CA2176248A1 - Methods and means for determining the female fertile period - Google Patents
Methods and means for determining the female fertile periodInfo
- Publication number
- CA2176248A1 CA2176248A1 CA002176248A CA2176248A CA2176248A1 CA 2176248 A1 CA2176248 A1 CA 2176248A1 CA 002176248 A CA002176248 A CA 002176248A CA 2176248 A CA2176248 A CA 2176248A CA 2176248 A1 CA2176248 A1 CA 2176248A1
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- Canada
- Prior art keywords
- day
- period
- hormone
- fertile period
- fertile
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/743—Steroid hormones
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16H—HEALTHCARE INFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR THE HANDLING OR PROCESSING OF MEDICAL OR HEALTHCARE DATA
- G16H40/00—ICT specially adapted for the management or administration of healthcare resources or facilities; ICT specially adapted for the management or operation of medical equipment or devices
- G16H40/60—ICT specially adapted for the management or administration of healthcare resources or facilities; ICT specially adapted for the management or operation of medical equipment or devices for the operation of medical equipment or devices
- G16H40/63—ICT specially adapted for the management or administration of healthcare resources or facilities; ICT specially adapted for the management or operation of medical equipment or devices for the operation of medical equipment or devices for local operation
Abstract
Provided is a highly reliable method to predict the beginning and/or ending of the fertile period for a female for each menstrual cycle. The methods and means provided advantageously address the day-to-day, cycle-to-cycle, and women-to-women variability in fertility hormone levels by analyzing the measurements of serial hormone concentrations in the midst of daily hormonal variations to determine when an actual and significant increase in the concentration of the monitored hormone has begun. Thus the present disclosure is directed to a method that combines existing hormone assay methods with calculation procedures to optimize the predictive values of daily hormonal changes. In this way a reliable and useful prediction of the fertile period is achieved with the concomitant assurance that the beginning and/or end of the fertile period has been reached so that such declarations are sufficient to provide both fertility enhancement and contraceptive utility.
Description
WO95/16920 ~ 4 8 PCT/US94/14455 METHODS AND MEANS FOR DETERMINING THE FEMALE FERTLE
PERIOD
Field of the Invention The present invention relates generally to methods and means for d~t~ll. inillg S periodic fertility of a female. More particularly, the present invention utilizes methods and means for d~lell- iri--g when the fertile period begins via the evaluation of a series of Ille~ulc~lllents of urine hormones. The measurements are made with regular assay methods, escenti~lly daily, after the first day of menses and con~ lg until the end of the fertile period. In addition, the present invention relates to a method for verifying the 0 actual occurrence of ovulation, and therefore, the end of fertile period.
Background of the Invention Methods for modifying human female fertility There are several methods to modify human female fertility. Pharmacological agents can be utiliæd to modify either the endocrinological sequence or the viability of 15 the gametes to reduce fertility. Examples are ovulation inducing drugs that çnh~nce fertility and oral contraceptive and spermicides that reduce fertility. Mechanically, barriers such as condoms and diaphragms can be used during sexual intercourse to prevent unwanted pregnancy. Another widely practiced method in modifying fertility is the timing method. The timing method is based on analysis provides an estimate of 20 when ovulation is likely to occur and thus sexual activity can be scheduled to maximize or minimi7P the probability of conception. Pharmacological and ml~h~nic~l methods are beyond the scope of this invention as the present invention is directed to increasing the effectiveness of the timing method.
25Timing ovulation There are several known methods for approxim~ting when ovulation has occurred, some of which are less reliable than others. Conventional methods for e~"-,;nil~g the fertile period, or for predicting the day of ovulation, range from tracking dates on a calendar to methods involving self-ex~min~tion and the charting of body 2l-76248 WO 95/16920 PCr/US94/1445 tt;lll~lCld~UlC (sympto-thermal method).
The c~l~f~ r method is based on the ~etefmin~tion of the usual length of a woman's menstrual cycle and dividing by two. The ~lcsulll~ion is that ovulation occurs approximately in the middle of the cycle. For example, if the woman's normal 5 cycle length is 23-24 days, ovulation is presumed to occur on the 12th day; if the woman's nommal cycle length is 40 days, ovulation may be presumed to occur between days 20 and 28. This method only provides a rough estim~te of when ovulation mayoccur and is very liffi~ult to use for most women whose cycle length varies from cycle to cycle.
Altematively, a woman may know approximately when she is ovulating if she experiences a mid-month sharp pain in the lower abdomen, inrlic~ting that ovulation is about to, or has just occurred. However, this is not a very reliable sign as this pain can be confused with other pain and is not felt by every woman.
The sympto-thermal method increases the indicative value of the c~len-~r method, and is based on the theory that basal body Ltlll~ela~ulc rises discemibly (about 0.4 C) shortly after ovulation has occurred. This phenomenon is related to the production of pro~,t~,one by the Graffian follicle. Rec~nce ovulation usually occurs around the middle of the menstrual cycle, basal body telllpeld~ulc monitoring is begun at about that time. To detect the increase in telllpcldlulc, a woman must take her telllpcldlulc every morning, on waking, at the same time throughout the cycle and record the results, typically as a graph of telll~ld~ulc versus day of cycle. When an a~plupliately sensilive th~l,llulllc~er is plupclly used, a distinct increase in basal body ~tlllpcldtLIc should be evident after ovulation has occurred. After lccordhlg this infommation over several months, a woman may be able to predict when ovulation will occur in subsequent cycles. However, as stated above, this method, is unsuitable for women who have irregular cycle length.
These methods of appro~im~ting day of ovulation have several disadvantages. A
fim~ problem is the difficulty in ~lcdi-;~illg the length of the ongoing menstrual cycle from previous cycles because most women do not have a standardized cycle length _ WO 95116920 ~ 1 7 6 2 ~ ~ PcrluS94/14455 for every cycle. When used as a method for preventing conception, each of these methods involves long intervals when users must practice ~hstin.onre or use additional contraceptive means. These methods may also involve ambiguity in hl~ c~illg the inriir~tor of the beginning of the fertile period, as well as a high probability of method S failure, i.e., conccl)lion. For example, the conventional sympto-thermal method of collL~ ion involves an abstin~nre period or risk interval of ten days and has an efficacy (Pearl Index) of 22 pregn~nries per 100 women-year of use. (Fertility &
Sterility, 36: 152 [1981]; 36: 591 [1981]; 40: 773 [1983] and 44: 328 [1985].) Furthermore, the women in these studies were closely monitored, and therefore, the 10 reported rate of pregnancy is probably the "i";"""" rate to be expected when using the sympto-thermal method. The sympto-thermal method has an efficacy similar to the high end of the range of efficacy reported for users of barrier-type (condom, diaphragm) contraceptives .
Ovulation prediction by horrnnne assay J.P. Royston (Biometrics, 38, 397-406, 1982) describes the relationship among basal body tclll~cl~thlc, ovulation, and the risk of conccy~ion~ with special reference to the lifetimes of sperm and egg.
A review article by the World Health Organization Task Force (Int. J. Fertil.
30(3), 18-30, 1985) describes a ~ ,e-;live multi-center study to develop universal immunochclllical tests for pre~licting the fertile period in women.
Brown et al. (Am. J. Obstet. Gynecol. 157,1082-1089, 1987) describes a method for d~t~ ",;nin~ a woman's fertility period based on the l,le&ulcnlenl of estrogen and PDG in urine with an enzyme-immnno~cc~y method.
Brown et. al. (Int. J. Gynecol. Obstet, Suppl. 1, 111-122, 1989) describes the hll~ol~lce of urinary estrogen and pregn~n~-liol llleasul~lllcn~ in identifying the period of fertility and infertility during the lllCll:~ll ual cycle. They also describe an immunoassay based on reagents coated onto walls of a mo(li~d cuvette.
J.P. Royston (Statistics in Medicine, 10, 221-240, 1991) reviews recent wo 95tl6920 2 1 ~ ~ 2 4 8 pCrlUS94/1445 progress on the use of statistics in identifying the fertile phase of the human menstrual cycle. Methods using threshold, or Bayesian change-point, and CUSUM (cumulative sum) techniques are diccucsed Blackwell and Brown (Steroids, 57: 554-562, 1992) discuss immunochemical 5 assay procedures and suggest to apply time series analysis for the recognition of increase in urinary estrogen as a marker for the beginning of the potential fertile period.
U.S. Patent No. 5,118,630 describes a method whereby an increase in pregnanediol glucuronide concentration in urine is cc,~ t;d to a threshold as a means to confirrn ovulation.
WIPO Patent Publications WO 94/04924, WO 94/04925, WO 94/04926 and WO 94/04928 describe a method whereby hormones are measured, and along with information about the woman's prior cycles, are used to predict ovulation in the current cycle.
Recently, several methods have been made available to either predict the time of15 ovulation or to confirm its oc~;u,l~nce. These m~thorlc are typically based on changes in ~çtçct~ble levels of urine hormones. Urinary hormone concentrations vary throughout the menstrual cycle. For example, ovulation is generally preceded by an increase in urinary estrogen-type hormones. The level of luteinizing hormone ("LH"), increases .cignifir~ntly the day before ovulation occurs. In many tests, the increased LH
20 conce..l.alion is intiic~t~ by a color change on a dipstick when contacted with a urine sample. Because the test result predicts ovulation before it has happened, it may provide a female with a better chance of conceiving as compared to the traditional methods that indicate that ovulation has already occurred. However, these ovulation prediction tests still have several disadv~nt~ges When used for co..ll~c~li~/e ~ oses these tests must 25 be performed very close to the time of ovulation since detectable LH increase occur only 24 hours before ovulation and LH levels return to normal in about 24 hours afterovulation. As stated above, it is not easy to predict the present cycle based on the history of previous cycles for women with irregular cycle lengths. A second drawback for the LH tests is that the strong surge in conc~ ,alion happens too close to the Wo 95116920 21 ~ ~ 2 ~ 8 PCT/US94/14455 s ovulation for contraceptive use. For contraceptive use, the LH test detects hormonal changes at a time too close to ovulation and therefore generally not sufficiently reliable.
Furthermore, the LH surge does not always occur, rendering the ~f t~- ,.,in~tion of fertile period ploble11ldtic.
As stated above, it is known that levels of proge~elone are elevated shortly after ovulation, see e.g. U.S. Patent No. 5,118,630. A major disadvantage of this method is that it can only confirm ovulation. Therefore, it is of no use for either conception enh~nrement or co-1L,a~ tive purposes, as it can only de~,,.l i.le the post-ovulation infertile period, not the pre-ovulation infertile period. Again, the method relies on the previous cycle length to predict when ovulation is likely to occur so that a woman can start testing PDG levels. For these reasons, it would be beneficial to provide a method by which the beginning and/or end of the fertile period could be predicted reliably from hormone concentration measulc-l~f ~1~, without having to rely on unreliable timing methods or measu-~-llents.
Thus, there is a co"l ;~-~l ;ng need to provide a highly reliable method to predict the beginning, and/or the end, of a female's fertile period. In addition, a need exists to provide such a method which addresses the day-to-day, cycle-to-cycle, and woman-to-woman variability in fertility hormone levels. Furthermore, such a method should be preferably provided in a form and at a cost which is suitable for in-home use by an 20 unskilled person.
The present invention provides a method that combines existing hormone assay methods with c~lcnl~tion procedures to ~~ li~ the predictive values of daily hormonal ch~rl~s In this way a reliable and useful prediction of the fertile period is achieved with the concollli~-L assurance that the beginning and/or end of the fertile period has 25 been reached so that such declarations are sllffirie.nt to provide both fertility enh~nrP~nP~,lt and conL~ ive utility.
WO95/16920 ~ 2~8 PCr/US94/14455 Summary of the Invention In one aspect, the invention provides a method for dclcllllhlillg the periodic fertile period in a female, the method including the steps of:
(a) ...e~ufi..g a hormone level in a daily sample of a biological fluid of the 5 female for a testing period commencing on a fixed number of days from the start of menses;
b) storing the measured hormone level and corresponding day from start of menses into a data storage means;
(c) analyzing the hormone level using a cc,---~!u~lional means, the co---l.u~lional 10 means comprising a miclupl.~cessor for applying a fitting function to norm~li7~d data;
(d) ~luces~illg analyzed data in a regime of three stages, including a baseline stage, a pre~icting stage and a conr"l"i,.g stage;
(e) extrapolating an expected horm- n~- level from the fitted function;
(f) the predicting stage co",~ ;"g .,.ea~u,cd hormone level to extrapolated 15 hormone level using a scoring means, the scoring means defining a preset relationship between "-ea~u.cd and extrapolated hormone values;
(g) decign~ting the first day of the fertile period as the day when accumnl~ttod scores in the predicting stage reach or exceed a predet,_ll.,i..ed value;
(h) the co--fi.."ing stage storing the mesured hormone level from the first day of the fertile period until the end of the testing period;
(i) analyzing the stored hormone data at the end of the testing period with a second scoring means comprising a preset decision rule;
(j) ~l.o,cign~ting the last day of the fertile period based on the oulcol"e of the second scoring means; and (k) displaying the first and last days of the fertile period on an output means;
whereby the female is considered capable of concc~lion from the time of reaching the first day of the fertile period until the last day of the fertile period.
The invention also provides a method for dctcll~linillg the first day of the fertile period WO 95116920 ;~ 1 7 6 2 4 8 PCr/US94/14455 The invention further provides a method for det~ -,;,-i,-g the final day of thefertile period.
The plcfc-l~,d method of the invention predicts both a first and a last day of the fertile period based on daily hormone measu.~,me~ ., most ~1cfe1dbly EIG, commencing a fixed number of days usually 1 to 8, pl~cfe1ably 2 to 5, most ~ulcfclably 5, from the start of menses. The daily hormone mea~.ulclllc,1L~7 continue for a fixed period of time, preferably lO days, from the start of testing.
Brief Description of the Figures Figure 1 shows a hard-held meter suitable for use in the method of the invention.
Figure 2 shows a disposable self-performing immunochromatography strip suitable for use in the method of the present invention.
Detailed Description of the Invention The present invention relates to an improved method for predicting the day of ovulation by the prospective estimation of a future estrogen holl.1one level. Thus, in one aspect, the invention provides a method for monitoring an optimal period for concc~tion. In addition, based on the prospective analysis of urinary progc~.lc1one hormone I11e~.ul~illlcllL~., the present invention provides verification that ovulation has occurred. Therefore, in another aspect, by early prediction and verification of ovulation the invention provides a method for conception prevention.
Broadly, one method of the invention is begun by observing the first day of menses in the menStrual cycle. Starting from a fixed day of the menstrual cycle,preferably day l through day 8, the level of estrone is 1lled..ulcd and the results recorded 25 on a daily basis. Any number of mea..ulcllle~ll or assay methods known in the art may be used. However, an illl~JUlLalll criterion for choosing the me~.ulelllent or assay method is that the Ille~.ul~,lllcnL of the hormone marker should be at least precise enough to be semi~u~,LiLdLi~e. The result is entered into a c~lc~ tion means which stores the result and l~c~fo1~s calculations, p~eft,~bly on a daily basis, as will be described in Wo 95/16920 ~ 3 2 ~ - PCTIUS94/1445~
., more detail below. The results of the first few days of hormone mea~,ulcl,lent thus serve as a baseline which is calculated for each female and, therefore reflects cycle variability unique to the user. When the progression of the change in estrone concellLIdlion meets the criteria set by the present invention, the beginning of fertile period is declared. From S that day, the daily Illeasulel.,t;lll of estrone ceases and a daily Illedsul~lllt;lll of progeslelone may begin. Again, proge~,Lerone may be measured following techniques known in the art as long as the method is suitable for at least semi-qu~ e analysis.
These results are also stored and analyzed by a c~lc~ tion means. When the change in progesterone concentration meets the criteria set by the present invention, the end of the l O fertility period is declared.
Briefly, then, a method of the invention comprises the following elemt-nt.s:
a) daily m~,u~ ;llls of the concellll~,lion of one or more fertility hormones in a biological sample;
b) a data storage means in which to input the hormone concentration data;
c) colll~uLaLional capacity for no~n~li7ing the data and fitting a function;
d) an evaluation means for extrapolating the fitted function to provide an esli...;1lr of the next day's hormone concellLIdLion;
e) a scoring means for decision making based on the relative slope of the fitted function and the colll~;,lison of the expected concellLIdLion to the observed concentration on the day of mea~,ult.ll~llL; and f) an output means to indicate the beginning of the fertile period or the end of the fertile period.
The result of the strip assay is read by the hand-held meter which also stores the result and p~,l~lllls c~lrul~tions, as will be ~rsrrihecl in more detail below. The improved perf~ e of the present invention over the other methods known in the art is due in part to the regime-like m~thrm~tir,~l scheme tailored for this particular use which can reliably predict i.--~ ovulation. F..~senti~lly, the method utilizes a baseline region, a running window, a set of st~ti~tic~l decision making rules, and testing of a fixed duration. A baseline region is c~lr~ trd and used to study the variation of the Wo 95/16920 2 t 7 6 2 ~ 8 PCT/US94/14455 observed hormone level in the present menstrual cycle itself as basis for later St~tictir~l decision. Each menstrual cycle uses its own baseline without the influence of or ~cfelcnce to previous menstrual cycles or other individuals. This simplifies operation and makes the method in~P.pen~lPnt of cycle-to-cycle variability within the same 5 individual. A running window scheme maximizes the efficiency and improves the sensitivity to .~ignific~nt changes in hormone levels. The st~ticti~al rules enhance the validity of decision making and the fixed duration of testing maximizes the efficiency of operation while .,.i.~i...;~.i-.g expense and labor. The development and op~i"li;~alion of the m~thPm~tir~l scheme will be described in more detail below.
An alternate and plcPcllcd method of the invention begins by observing the first day of menses in the menstrual cycle. From a pre-selected fixed day, preferably day 5, the level of estrone is assayed and the results recorded on a daily basis. For assaying urinary steriod hormone, any number of assay methods known in the art may be used, but again, an hlll)o"anl criterion for choosing the assay method is that the meas.llclllcl t 15 of the hormone marker should be at least precise enough to be semi-qu~ntit~tive. As before, the level of estrone in the first few days measulclllcn~. serve as a baseline unique for the individual female. When an increase in hormone concentration meets the criteria described herein, the beginning of the fertile period is inrlic~tPcl to the user.
Again, any number of assay methods known in the art may be utilized in the 20 method of the invention. In making the selection, an hllpol ~1t criteria is that the assay provide at least a semi-qu~ ivc result (as opposed to a qualitative, i.e.7 "yes/no"
result) be "user-friendly" and u~lco...l)lir~erl That is, the assay should be sutiable for in-home use by a relatively untrained person. Immunoassays developed for use on dip sticks, membranes, strips, etc. are thus plcfe.lcd. Most plc~ lcd is a disposable, self-25 performing solid-phase immunoassay strip which, upon use, may be scanned by a rP-flect~nre reader as described below. The use of imm~lnrJclllulllalography test stips is well known to those skilled in the art. F.~peci~lly suitable for strips useful in the present invention are those employing colloidal lllicrùp~licles. In this regard, the preferred llliclu~u~icle is described in U.S. Patent 5,252,459 to Tarcha, et al. The '459 patent WO 95/16920 PCr/US94/1445~
~17~2408 describes an indicator reagent, method and test kit for the analysis of an analyte in a test sample.
As mentioned above, the end result of the hormone mea~u,c"lc;lll must be nt rm~li7~1, fitted to a function to create a baseline against which subsequent hormone S lue~u~c~ "~ will be co.l.p~d. Such techniques can be performed m~n~ y following the method described infra, however, it is a preferred feature of the invention to utilize an es~enti~lly automated a~d~us which performs all steps of the method which involve measu,~",e--L, calculations, storage, etc. Thus, an additional and preferred feature of the invention is that the method of the present invention, when the calculation, 10 storage, timing, etc. functions are handled by a mic.ùp,ocessor-based reader, requires no input, or h~ ,ct~Lion of results, by the user. Indeed, when such a reader is utilized in the present invention, the hlrullllalion displayed by the reader is a signal or readout inllir~ting that the fertile period has been reached. Thus, there is no q~l~ntit~tive or semi-4~ /e result which is displayed for the user. This greatly simplifies the method 15 from the standpoint of the user.
Thus, when the progression of the change in estrone concentration meets the criteria set by the present invention, the begi.--1i..g of fertile period is declared. From that day, the daily "Rasu,~"ænl of estrone estrone continues until a pre-selected fixed day (e.g. day 14) for the end of monitoring At the end of the monitoring period, the 20 meter processes the stored results of the estrone level using the criteria set by the present invention to predict and declare the end of the fertility period. The improved perfo"nal1ce of the present invention over the other methods known in the art is due in part to the invented regime-like m~th~m~ti~l scheme tailored for this particular use to predict illllll;ll~l~t ovulation. F~senti~lly, the m~th.-m~tir~l scheme consists of a baseline 25 region, a running window, a set of st~ti~tic~l decision making rules, and a fixed test duration. A baseline region is used to study the variation of the observed hormone level in the present -ænsllual cycle itself as basis for later st~ticti~l decision. Each menstrual cycle uses its own baseli-le without the influence of previous ~ ual cycles or other individuals. This simplifies operation and avoids complication that no two cycles of - WO95/16920 ~ L 7 ~ 2 4 8 PCT/US94/14455 the same individual are i~lPntir~l A running window scheme maximizes the efficiency and improves the sensitivity to detect a cignifir~nt change in the hormone level. The st~tcitic~l rules enh~nre the validity of decision m~king The fixed duration of testing maximizes the efficiency of operation and provide user with friendly operation system.
S The development and o~tillliz~ion of the m;.ll,r",~lir~l scheme will be described in more detail below.
In a preferred method of practicing the invention, a hand-held meter is described for use in accordance with the invention. Figure I illustrates a hand-held meter suitable for use in fertility period monitoring. Figure 2 shows a strip for solid state 10 immunoassay itself. Referring now to Figure 1, the meter I comprises a top casing 104 and a bottom casing 110 with an opening 109 for receiving the carrier 100. A test strip is held in place on carrier 100 by ridge 101 and locating pin 103. When placed in the meter, gear 102 of carrier 100 engages with the driving gear train of a motor inside the reader. An optical refl~ct~nce read head is housed inside the reader depicted as a 15 protrusion 105 of the meter. Assay results as well as user inputs are displayed on a liquid crystal display 108. Two buttons or switches 106 and 107 provide control and user interface to the meter. Referring now to Figure 2, the solid phase immunoassay strip 2 is ~u~poll~d by a plactic carrier 200 which has a notch 201 comple.llc;llli~y to locking pin 103 on carrier 100 to allow reproducible positioning. A reagent pad 202 20 cont~inc, e.g. a diffusible labeled reagent, and is in contact with nitrocellulose strip 203 which co~Lains an immobilized capture reagent for immuno reaction. When the user applies a urine sample to reagent pad 202, the reagent label reacts with steroid in the sample and the complex will be llanspolLed along the nitrocellulose strip by capillar action across the capture reagent which immobilizes the label-steroid complex. After an 25 ~ lU~lidtl, development time, the intensity of the signal at the capture reagent can then be recorded with a sc~nning reflect~nre reader (inside the meter under the read head) 105. ~,fe,dbly, the meter is a battery powered device to enh~nre the portability and convenience of use of the whole system.
The hand-held meter is controlled by a mi~;lul,locessor. Optical elements consist Wo 95/16920 2 ~ 6 2 ~ ~ PCr/US94/1445S
of illumination and photo sensors which function to provide a data string as a reflection reading to a voltage-to-frequency converter for result signals to the processor. A motor provides sc~nning motion under control of the processor. A real time clock keeps track of timed parameters, e.g., day of testing and day of menstrual cycle. The user interface 5 driver relays the information from and to the processor with the user through a display and control buttons. Batteries and a power supply controller ensures sufficient power that an entire menstrual cycle may be monitored before battery repl~e-m~nt is required.
A calibration data bank is also included and functions to convert optical signal to analyte concentration.
As would be understood by the skilled person, individual features may be replaced by equivalent on subst~nti~lly equivalent elements in order to achieve the same objective as outlined above. For example, discrete electronic parts could be replæed with an application specific integrated circuitry (ASIC) and visa versa.
1 5 Definitions The following definitions are applicable to the present invention. ~th.om~tic~l operations are also explained when a~lupliately associated with the definition.
The term "fitting function" refers to a mathematical expression of the relationship between the day of the menstrual cycle (typically, day 1 through 28) and the 20 hormone con~entration that is Illea~,ured on that day. Although many fitting functions can be applied to the conc~ ion data, three especi~lly useful functions will be described which provide a high degree of certainty for the perfollllance of the present mventlon.
a. Linear Function. The first fitting function is a simple linear equation and 25 has the advantage of simplicity, fast c~lcul~tion time, and easy i~ ,le..,~"~ ion. The linear fitting function has the following formula:
concell~dtion = aO + (a1 x Day) where aO is the hltel-;ept of the linear fitting function, al is the slope of the linear fitting function and Day is the day of the IllensLl,lal cycle with Day=1 being the first day of ~l762~.8 menses. aO and a1 are numeric constants determined by the method of least squares.
The unit of concentration in this equation is that used in the collc~onding assay method. For example, nano-mole per liter is commonly used for assaying estrone 3-glucuronide (ElG) by immunoreaction. The linear least square plocedulG is easily 5 calculated and is well known or can be readily found in any number of reference texts.
e.g., "FORTRAN Programs For Scientists and Engineers", by Alan R. Miller (Sybex Inc., San Francisco (1982).
b. Linear Inverse Exponential Function. The second preferred function is a non-linear one and has the following formula:
concentration = ( aO + (al x Day))/ (I + l/lO(a2+ Day)) where a~, al and Day are as defined above, and a2 is another numeric consk~t determined by the method of least squares. This non-linear fitting function is more 15 complex since the cA~oncl.Lial CollllJoll."ll~ require more Culll~uLillg power and time to execute. However, with its complex nature, it provides an oppollullily to develop the adaptive algorithm to be ~ cucsed below. The llulllelic col.~lal1t~ of the function are estim~tr~l by the method of least squares to provide a -,;.,;"""" squared difference between the concclllldlion predicted by the fitting function and the actual concc--L,dlion 20 observed. For the non-linear fitting function, a2 is the exponential modulus. The use of this function is more complex and usually involves multi-variable o~l;,,,;7i~'ion iterative search. The Simplex method is one such well known method and is suitable for use in the method of the invention. A detailed rlc srnrtion of the Simplex method can be found in standard linear ~l.~l,.~lllll;l~g ~lobl_.ll solving texts such as "Mathematical 25 Progr~,u,.ir~g'', by Reinfeld and Vogel, Published by Prentice-Hall, Inc. Englewood Cliffs, N.J. (1958). Briefly, the Simplex method iflpnti~lps a basic feasible solution for the con~lldilled multi-variable equations. Then, the method is used to dcl~ ,--;-,e whether an (~)~illlUIII value has been reached by ~ub~l~clillg the value c~lcul~trd from the fitting model for each basis point from the actual analytical value at the point. This dirrclcllce Wo 95/16920 ~ 1 ~ 6 2 ~ 8 PCr/US94/14455 is squared and the squared difference is used to rank the sets of basis parameters. If an Op~illlUIII has not been reached, the method provides a way of moving to a new solution that will have a better value of the objective function. In the method of the invention, for example, four observed hormone concentrations on four consecutive days are set up as S four .cimlllt~nrous equations with a set of starting estim~tPd values of aO, al, and a2 with 4 slack variables, one for each of the four equations in a tableau. These variables are tested to see whether all the net gains are negative. If it is, then the OpLh~u~ has been found. If not, the operation of pivoting is carried out using the most marginal net gain variable as the pivot element and removing the variable with the least positive net l O gain to arrive at a new basic feasible solution with modified variables. The process is repeated until the o~Lilllulll has been found. The results are a set of values for the fitting function that give least square deviation for the observed hormone concentration.
c. Arit~unetic Average Function. The third function is a simple ~, ;LI ,., If ~liC
average c~lcul~tion. By using average values from a selected range of days as a baseline 15 to set up threshold values for comr~ricon to the observed value of a later day, a mathematical relationship is set up. This function is the most simple to use but not very reliable if used to predict beginning of the fertile cycle because it is more sensitive to data variability than the previously descrihecl two functions. However, using a sophisticated function for prediction of the be~ g of the fertile period to st~tictir~lly comren.c~te 20 for the data variation difficulties, such as the linear equation or non-linear equation as described above, a simple method could be used for predicting the end of the fertile period.
The term "slope" refers to the rate of change in hormone concentration. For the linear fitting function, it is equivalent to the value of a1. For the non-linear fitting 25 function, it is the evaluation of the first derivative of the fitting function.
The term "relative slope" refers to the value of slope mllltirlird by 100 then divided by the average values of the obse, ~d hormone conce"ll~Lion in the fitting function. In other words, relative slope is the average concentration norm~li7~d rate of p~"ce~L~ge concentration change. Since function fitting is done on the data in base ~1762~
- wo 95/16920 ~ ~ PCT/US94/14455 window, the slope is calculated from data in base window. [Define 'Base Window']The use of relative slope instead of slope itself reduces sensitivity of decision making to the individual to individual variation.
The term "predicted value" refers to the threshold value of hormone 5 conce~LIdLion, based on the fitted function, e~t~tiing one day beyond the fitting period.
For example, if a four day fitting window is used the predicted value of today would the evaluation of the fitted function with the variables least square fitted from the observed hormone concentration of days from 4 days ago to yesterday.
The term "threshold value" refers to the value of the sum of predicted value and10 the value of fitting standard deviation multiplied by a chosen multiplier. By co",l.~. ;llg the threshold value to the observed hormone concentration, the mathematical processes in the present invention de~e~ P whether a change in the trend in the hormone conce..l.d~ion has occurred. The multiplier is similar to a st~tictic~ 't' test and is selected using similar considerations in that the multiplier ~'~csessçs~ the confiflPn~e level to be given an ~csçs~,nr,.l that an observed change in hormonal concellllalion warrants an action. For the third fitting function of simple average as described above, the threshold value is the product of "baseline value" with a pre-set multiplier.
The term "data variability" refers to that property of the concentration ~e~u~ L~ that reflect a female's own day to day hormone concenLIdLion changes and result in minor deviations from a smooth slope. For example, variability can be caused by the changes in the way the hormone is metabolized in liver, the rate of hormone secretion into the kidney, the volume of liquid secreted by kidney, the conce"lld~illg process of urine in the urinary bladder, and variations in the assay method used to quantify the hormone conc~LI~tion.
The term "b~cPIinP" refers to the hormone concellL,dLions as nle~ulc;d on several col-~e~-uli-.re days that are characterized by relatively low values. Thel.,folc, the baseline values collc~L~ond to days where an increase of ovarian activity is unlikely. Typically, b~cPIinP for ElG l~,~lcse~ day 2 to day 5 of each cycle while the b~cPIinP for PDG
Ic~lcsellL~ the first 4 days of PDG measu,e,l,t;,ll of each cycle.
PERIOD
Field of the Invention The present invention relates generally to methods and means for d~t~ll. inillg S periodic fertility of a female. More particularly, the present invention utilizes methods and means for d~lell- iri--g when the fertile period begins via the evaluation of a series of Ille~ulc~lllents of urine hormones. The measurements are made with regular assay methods, escenti~lly daily, after the first day of menses and con~ lg until the end of the fertile period. In addition, the present invention relates to a method for verifying the 0 actual occurrence of ovulation, and therefore, the end of fertile period.
Background of the Invention Methods for modifying human female fertility There are several methods to modify human female fertility. Pharmacological agents can be utiliæd to modify either the endocrinological sequence or the viability of 15 the gametes to reduce fertility. Examples are ovulation inducing drugs that çnh~nce fertility and oral contraceptive and spermicides that reduce fertility. Mechanically, barriers such as condoms and diaphragms can be used during sexual intercourse to prevent unwanted pregnancy. Another widely practiced method in modifying fertility is the timing method. The timing method is based on analysis provides an estimate of 20 when ovulation is likely to occur and thus sexual activity can be scheduled to maximize or minimi7P the probability of conception. Pharmacological and ml~h~nic~l methods are beyond the scope of this invention as the present invention is directed to increasing the effectiveness of the timing method.
25Timing ovulation There are several known methods for approxim~ting when ovulation has occurred, some of which are less reliable than others. Conventional methods for e~"-,;nil~g the fertile period, or for predicting the day of ovulation, range from tracking dates on a calendar to methods involving self-ex~min~tion and the charting of body 2l-76248 WO 95/16920 PCr/US94/1445 tt;lll~lCld~UlC (sympto-thermal method).
The c~l~f~ r method is based on the ~etefmin~tion of the usual length of a woman's menstrual cycle and dividing by two. The ~lcsulll~ion is that ovulation occurs approximately in the middle of the cycle. For example, if the woman's normal 5 cycle length is 23-24 days, ovulation is presumed to occur on the 12th day; if the woman's nommal cycle length is 40 days, ovulation may be presumed to occur between days 20 and 28. This method only provides a rough estim~te of when ovulation mayoccur and is very liffi~ult to use for most women whose cycle length varies from cycle to cycle.
Altematively, a woman may know approximately when she is ovulating if she experiences a mid-month sharp pain in the lower abdomen, inrlic~ting that ovulation is about to, or has just occurred. However, this is not a very reliable sign as this pain can be confused with other pain and is not felt by every woman.
The sympto-thermal method increases the indicative value of the c~len-~r method, and is based on the theory that basal body Ltlll~ela~ulc rises discemibly (about 0.4 C) shortly after ovulation has occurred. This phenomenon is related to the production of pro~,t~,one by the Graffian follicle. Rec~nce ovulation usually occurs around the middle of the menstrual cycle, basal body telllpeld~ulc monitoring is begun at about that time. To detect the increase in telllpcldlulc, a woman must take her telllpcldlulc every morning, on waking, at the same time throughout the cycle and record the results, typically as a graph of telll~ld~ulc versus day of cycle. When an a~plupliately sensilive th~l,llulllc~er is plupclly used, a distinct increase in basal body ~tlllpcldtLIc should be evident after ovulation has occurred. After lccordhlg this infommation over several months, a woman may be able to predict when ovulation will occur in subsequent cycles. However, as stated above, this method, is unsuitable for women who have irregular cycle length.
These methods of appro~im~ting day of ovulation have several disadvantages. A
fim~ problem is the difficulty in ~lcdi-;~illg the length of the ongoing menstrual cycle from previous cycles because most women do not have a standardized cycle length _ WO 95116920 ~ 1 7 6 2 ~ ~ PcrluS94/14455 for every cycle. When used as a method for preventing conception, each of these methods involves long intervals when users must practice ~hstin.onre or use additional contraceptive means. These methods may also involve ambiguity in hl~ c~illg the inriir~tor of the beginning of the fertile period, as well as a high probability of method S failure, i.e., conccl)lion. For example, the conventional sympto-thermal method of collL~ ion involves an abstin~nre period or risk interval of ten days and has an efficacy (Pearl Index) of 22 pregn~nries per 100 women-year of use. (Fertility &
Sterility, 36: 152 [1981]; 36: 591 [1981]; 40: 773 [1983] and 44: 328 [1985].) Furthermore, the women in these studies were closely monitored, and therefore, the 10 reported rate of pregnancy is probably the "i";"""" rate to be expected when using the sympto-thermal method. The sympto-thermal method has an efficacy similar to the high end of the range of efficacy reported for users of barrier-type (condom, diaphragm) contraceptives .
Ovulation prediction by horrnnne assay J.P. Royston (Biometrics, 38, 397-406, 1982) describes the relationship among basal body tclll~cl~thlc, ovulation, and the risk of conccy~ion~ with special reference to the lifetimes of sperm and egg.
A review article by the World Health Organization Task Force (Int. J. Fertil.
30(3), 18-30, 1985) describes a ~ ,e-;live multi-center study to develop universal immunochclllical tests for pre~licting the fertile period in women.
Brown et al. (Am. J. Obstet. Gynecol. 157,1082-1089, 1987) describes a method for d~t~ ",;nin~ a woman's fertility period based on the l,le&ulcnlenl of estrogen and PDG in urine with an enzyme-immnno~cc~y method.
Brown et. al. (Int. J. Gynecol. Obstet, Suppl. 1, 111-122, 1989) describes the hll~ol~lce of urinary estrogen and pregn~n~-liol llleasul~lllcn~ in identifying the period of fertility and infertility during the lllCll:~ll ual cycle. They also describe an immunoassay based on reagents coated onto walls of a mo(li~d cuvette.
J.P. Royston (Statistics in Medicine, 10, 221-240, 1991) reviews recent wo 95tl6920 2 1 ~ ~ 2 4 8 pCrlUS94/1445 progress on the use of statistics in identifying the fertile phase of the human menstrual cycle. Methods using threshold, or Bayesian change-point, and CUSUM (cumulative sum) techniques are diccucsed Blackwell and Brown (Steroids, 57: 554-562, 1992) discuss immunochemical 5 assay procedures and suggest to apply time series analysis for the recognition of increase in urinary estrogen as a marker for the beginning of the potential fertile period.
U.S. Patent No. 5,118,630 describes a method whereby an increase in pregnanediol glucuronide concentration in urine is cc,~ t;d to a threshold as a means to confirrn ovulation.
WIPO Patent Publications WO 94/04924, WO 94/04925, WO 94/04926 and WO 94/04928 describe a method whereby hormones are measured, and along with information about the woman's prior cycles, are used to predict ovulation in the current cycle.
Recently, several methods have been made available to either predict the time of15 ovulation or to confirm its oc~;u,l~nce. These m~thorlc are typically based on changes in ~çtçct~ble levels of urine hormones. Urinary hormone concentrations vary throughout the menstrual cycle. For example, ovulation is generally preceded by an increase in urinary estrogen-type hormones. The level of luteinizing hormone ("LH"), increases .cignifir~ntly the day before ovulation occurs. In many tests, the increased LH
20 conce..l.alion is intiic~t~ by a color change on a dipstick when contacted with a urine sample. Because the test result predicts ovulation before it has happened, it may provide a female with a better chance of conceiving as compared to the traditional methods that indicate that ovulation has already occurred. However, these ovulation prediction tests still have several disadv~nt~ges When used for co..ll~c~li~/e ~ oses these tests must 25 be performed very close to the time of ovulation since detectable LH increase occur only 24 hours before ovulation and LH levels return to normal in about 24 hours afterovulation. As stated above, it is not easy to predict the present cycle based on the history of previous cycles for women with irregular cycle lengths. A second drawback for the LH tests is that the strong surge in conc~ ,alion happens too close to the Wo 95116920 21 ~ ~ 2 ~ 8 PCT/US94/14455 s ovulation for contraceptive use. For contraceptive use, the LH test detects hormonal changes at a time too close to ovulation and therefore generally not sufficiently reliable.
Furthermore, the LH surge does not always occur, rendering the ~f t~- ,.,in~tion of fertile period ploble11ldtic.
As stated above, it is known that levels of proge~elone are elevated shortly after ovulation, see e.g. U.S. Patent No. 5,118,630. A major disadvantage of this method is that it can only confirm ovulation. Therefore, it is of no use for either conception enh~nrement or co-1L,a~ tive purposes, as it can only de~,,.l i.le the post-ovulation infertile period, not the pre-ovulation infertile period. Again, the method relies on the previous cycle length to predict when ovulation is likely to occur so that a woman can start testing PDG levels. For these reasons, it would be beneficial to provide a method by which the beginning and/or end of the fertile period could be predicted reliably from hormone concentration measulc-l~f ~1~, without having to rely on unreliable timing methods or measu-~-llents.
Thus, there is a co"l ;~-~l ;ng need to provide a highly reliable method to predict the beginning, and/or the end, of a female's fertile period. In addition, a need exists to provide such a method which addresses the day-to-day, cycle-to-cycle, and woman-to-woman variability in fertility hormone levels. Furthermore, such a method should be preferably provided in a form and at a cost which is suitable for in-home use by an 20 unskilled person.
The present invention provides a method that combines existing hormone assay methods with c~lcnl~tion procedures to ~~ li~ the predictive values of daily hormonal ch~rl~s In this way a reliable and useful prediction of the fertile period is achieved with the concollli~-L assurance that the beginning and/or end of the fertile period has 25 been reached so that such declarations are sllffirie.nt to provide both fertility enh~nrP~nP~,lt and conL~ ive utility.
WO95/16920 ~ 2~8 PCr/US94/14455 Summary of the Invention In one aspect, the invention provides a method for dclcllllhlillg the periodic fertile period in a female, the method including the steps of:
(a) ...e~ufi..g a hormone level in a daily sample of a biological fluid of the 5 female for a testing period commencing on a fixed number of days from the start of menses;
b) storing the measured hormone level and corresponding day from start of menses into a data storage means;
(c) analyzing the hormone level using a cc,---~!u~lional means, the co---l.u~lional 10 means comprising a miclupl.~cessor for applying a fitting function to norm~li7~d data;
(d) ~luces~illg analyzed data in a regime of three stages, including a baseline stage, a pre~icting stage and a conr"l"i,.g stage;
(e) extrapolating an expected horm- n~- level from the fitted function;
(f) the predicting stage co",~ ;"g .,.ea~u,cd hormone level to extrapolated 15 hormone level using a scoring means, the scoring means defining a preset relationship between "-ea~u.cd and extrapolated hormone values;
(g) decign~ting the first day of the fertile period as the day when accumnl~ttod scores in the predicting stage reach or exceed a predet,_ll.,i..ed value;
(h) the co--fi.."ing stage storing the mesured hormone level from the first day of the fertile period until the end of the testing period;
(i) analyzing the stored hormone data at the end of the testing period with a second scoring means comprising a preset decision rule;
(j) ~l.o,cign~ting the last day of the fertile period based on the oulcol"e of the second scoring means; and (k) displaying the first and last days of the fertile period on an output means;
whereby the female is considered capable of concc~lion from the time of reaching the first day of the fertile period until the last day of the fertile period.
The invention also provides a method for dctcll~linillg the first day of the fertile period WO 95116920 ;~ 1 7 6 2 4 8 PCr/US94/14455 The invention further provides a method for det~ -,;,-i,-g the final day of thefertile period.
The plcfc-l~,d method of the invention predicts both a first and a last day of the fertile period based on daily hormone measu.~,me~ ., most ~1cfe1dbly EIG, commencing a fixed number of days usually 1 to 8, pl~cfe1ably 2 to 5, most ~ulcfclably 5, from the start of menses. The daily hormone mea~.ulclllc,1L~7 continue for a fixed period of time, preferably lO days, from the start of testing.
Brief Description of the Figures Figure 1 shows a hard-held meter suitable for use in the method of the invention.
Figure 2 shows a disposable self-performing immunochromatography strip suitable for use in the method of the present invention.
Detailed Description of the Invention The present invention relates to an improved method for predicting the day of ovulation by the prospective estimation of a future estrogen holl.1one level. Thus, in one aspect, the invention provides a method for monitoring an optimal period for concc~tion. In addition, based on the prospective analysis of urinary progc~.lc1one hormone I11e~.ul~illlcllL~., the present invention provides verification that ovulation has occurred. Therefore, in another aspect, by early prediction and verification of ovulation the invention provides a method for conception prevention.
Broadly, one method of the invention is begun by observing the first day of menses in the menStrual cycle. Starting from a fixed day of the menstrual cycle,preferably day l through day 8, the level of estrone is 1lled..ulcd and the results recorded 25 on a daily basis. Any number of mea..ulcllle~ll or assay methods known in the art may be used. However, an illl~JUlLalll criterion for choosing the me~.ulelllent or assay method is that the Ille~.ul~,lllcnL of the hormone marker should be at least precise enough to be semi~u~,LiLdLi~e. The result is entered into a c~lc~ tion means which stores the result and l~c~fo1~s calculations, p~eft,~bly on a daily basis, as will be described in Wo 95/16920 ~ 3 2 ~ - PCTIUS94/1445~
., more detail below. The results of the first few days of hormone mea~,ulcl,lent thus serve as a baseline which is calculated for each female and, therefore reflects cycle variability unique to the user. When the progression of the change in estrone concellLIdlion meets the criteria set by the present invention, the beginning of fertile period is declared. From S that day, the daily Illeasulel.,t;lll of estrone ceases and a daily Illedsul~lllt;lll of progeslelone may begin. Again, proge~,Lerone may be measured following techniques known in the art as long as the method is suitable for at least semi-qu~ e analysis.
These results are also stored and analyzed by a c~lc~ tion means. When the change in progesterone concentration meets the criteria set by the present invention, the end of the l O fertility period is declared.
Briefly, then, a method of the invention comprises the following elemt-nt.s:
a) daily m~,u~ ;llls of the concellll~,lion of one or more fertility hormones in a biological sample;
b) a data storage means in which to input the hormone concentration data;
c) colll~uLaLional capacity for no~n~li7ing the data and fitting a function;
d) an evaluation means for extrapolating the fitted function to provide an esli...;1lr of the next day's hormone concellLIdLion;
e) a scoring means for decision making based on the relative slope of the fitted function and the colll~;,lison of the expected concellLIdLion to the observed concentration on the day of mea~,ult.ll~llL; and f) an output means to indicate the beginning of the fertile period or the end of the fertile period.
The result of the strip assay is read by the hand-held meter which also stores the result and p~,l~lllls c~lrul~tions, as will be ~rsrrihecl in more detail below. The improved perf~ e of the present invention over the other methods known in the art is due in part to the regime-like m~thrm~tir,~l scheme tailored for this particular use which can reliably predict i.--~ ovulation. F..~senti~lly, the method utilizes a baseline region, a running window, a set of st~ti~tic~l decision making rules, and testing of a fixed duration. A baseline region is c~lr~ trd and used to study the variation of the Wo 95/16920 2 t 7 6 2 ~ 8 PCT/US94/14455 observed hormone level in the present menstrual cycle itself as basis for later St~tictir~l decision. Each menstrual cycle uses its own baseline without the influence of or ~cfelcnce to previous menstrual cycles or other individuals. This simplifies operation and makes the method in~P.pen~lPnt of cycle-to-cycle variability within the same 5 individual. A running window scheme maximizes the efficiency and improves the sensitivity to .~ignific~nt changes in hormone levels. The st~ticti~al rules enhance the validity of decision making and the fixed duration of testing maximizes the efficiency of operation while .,.i.~i...;~.i-.g expense and labor. The development and op~i"li;~alion of the m~thPm~tir~l scheme will be described in more detail below.
An alternate and plcPcllcd method of the invention begins by observing the first day of menses in the menstrual cycle. From a pre-selected fixed day, preferably day 5, the level of estrone is assayed and the results recorded on a daily basis. For assaying urinary steriod hormone, any number of assay methods known in the art may be used, but again, an hlll)o"anl criterion for choosing the assay method is that the meas.llclllcl t 15 of the hormone marker should be at least precise enough to be semi-qu~ntit~tive. As before, the level of estrone in the first few days measulclllcn~. serve as a baseline unique for the individual female. When an increase in hormone concentration meets the criteria described herein, the beginning of the fertile period is inrlic~tPcl to the user.
Again, any number of assay methods known in the art may be utilized in the 20 method of the invention. In making the selection, an hllpol ~1t criteria is that the assay provide at least a semi-qu~ ivc result (as opposed to a qualitative, i.e.7 "yes/no"
result) be "user-friendly" and u~lco...l)lir~erl That is, the assay should be sutiable for in-home use by a relatively untrained person. Immunoassays developed for use on dip sticks, membranes, strips, etc. are thus plcfe.lcd. Most plc~ lcd is a disposable, self-25 performing solid-phase immunoassay strip which, upon use, may be scanned by a rP-flect~nre reader as described below. The use of imm~lnrJclllulllalography test stips is well known to those skilled in the art. F.~peci~lly suitable for strips useful in the present invention are those employing colloidal lllicrùp~licles. In this regard, the preferred llliclu~u~icle is described in U.S. Patent 5,252,459 to Tarcha, et al. The '459 patent WO 95/16920 PCr/US94/1445~
~17~2408 describes an indicator reagent, method and test kit for the analysis of an analyte in a test sample.
As mentioned above, the end result of the hormone mea~u,c"lc;lll must be nt rm~li7~1, fitted to a function to create a baseline against which subsequent hormone S lue~u~c~ "~ will be co.l.p~d. Such techniques can be performed m~n~ y following the method described infra, however, it is a preferred feature of the invention to utilize an es~enti~lly automated a~d~us which performs all steps of the method which involve measu,~",e--L, calculations, storage, etc. Thus, an additional and preferred feature of the invention is that the method of the present invention, when the calculation, 10 storage, timing, etc. functions are handled by a mic.ùp,ocessor-based reader, requires no input, or h~ ,ct~Lion of results, by the user. Indeed, when such a reader is utilized in the present invention, the hlrullllalion displayed by the reader is a signal or readout inllir~ting that the fertile period has been reached. Thus, there is no q~l~ntit~tive or semi-4~ /e result which is displayed for the user. This greatly simplifies the method 15 from the standpoint of the user.
Thus, when the progression of the change in estrone concentration meets the criteria set by the present invention, the begi.--1i..g of fertile period is declared. From that day, the daily "Rasu,~"ænl of estrone estrone continues until a pre-selected fixed day (e.g. day 14) for the end of monitoring At the end of the monitoring period, the 20 meter processes the stored results of the estrone level using the criteria set by the present invention to predict and declare the end of the fertility period. The improved perfo"nal1ce of the present invention over the other methods known in the art is due in part to the invented regime-like m~th~m~ti~l scheme tailored for this particular use to predict illllll;ll~l~t ovulation. F~senti~lly, the m~th.-m~tir~l scheme consists of a baseline 25 region, a running window, a set of st~ti~tic~l decision making rules, and a fixed test duration. A baseline region is used to study the variation of the observed hormone level in the present -ænsllual cycle itself as basis for later st~ticti~l decision. Each menstrual cycle uses its own baseli-le without the influence of previous ~ ual cycles or other individuals. This simplifies operation and avoids complication that no two cycles of - WO95/16920 ~ L 7 ~ 2 4 8 PCT/US94/14455 the same individual are i~lPntir~l A running window scheme maximizes the efficiency and improves the sensitivity to detect a cignifir~nt change in the hormone level. The st~tcitic~l rules enh~nre the validity of decision m~king The fixed duration of testing maximizes the efficiency of operation and provide user with friendly operation system.
S The development and o~tillliz~ion of the m;.ll,r",~lir~l scheme will be described in more detail below.
In a preferred method of practicing the invention, a hand-held meter is described for use in accordance with the invention. Figure I illustrates a hand-held meter suitable for use in fertility period monitoring. Figure 2 shows a strip for solid state 10 immunoassay itself. Referring now to Figure 1, the meter I comprises a top casing 104 and a bottom casing 110 with an opening 109 for receiving the carrier 100. A test strip is held in place on carrier 100 by ridge 101 and locating pin 103. When placed in the meter, gear 102 of carrier 100 engages with the driving gear train of a motor inside the reader. An optical refl~ct~nce read head is housed inside the reader depicted as a 15 protrusion 105 of the meter. Assay results as well as user inputs are displayed on a liquid crystal display 108. Two buttons or switches 106 and 107 provide control and user interface to the meter. Referring now to Figure 2, the solid phase immunoassay strip 2 is ~u~poll~d by a plactic carrier 200 which has a notch 201 comple.llc;llli~y to locking pin 103 on carrier 100 to allow reproducible positioning. A reagent pad 202 20 cont~inc, e.g. a diffusible labeled reagent, and is in contact with nitrocellulose strip 203 which co~Lains an immobilized capture reagent for immuno reaction. When the user applies a urine sample to reagent pad 202, the reagent label reacts with steroid in the sample and the complex will be llanspolLed along the nitrocellulose strip by capillar action across the capture reagent which immobilizes the label-steroid complex. After an 25 ~ lU~lidtl, development time, the intensity of the signal at the capture reagent can then be recorded with a sc~nning reflect~nre reader (inside the meter under the read head) 105. ~,fe,dbly, the meter is a battery powered device to enh~nre the portability and convenience of use of the whole system.
The hand-held meter is controlled by a mi~;lul,locessor. Optical elements consist Wo 95/16920 2 ~ 6 2 ~ ~ PCr/US94/1445S
of illumination and photo sensors which function to provide a data string as a reflection reading to a voltage-to-frequency converter for result signals to the processor. A motor provides sc~nning motion under control of the processor. A real time clock keeps track of timed parameters, e.g., day of testing and day of menstrual cycle. The user interface 5 driver relays the information from and to the processor with the user through a display and control buttons. Batteries and a power supply controller ensures sufficient power that an entire menstrual cycle may be monitored before battery repl~e-m~nt is required.
A calibration data bank is also included and functions to convert optical signal to analyte concentration.
As would be understood by the skilled person, individual features may be replaced by equivalent on subst~nti~lly equivalent elements in order to achieve the same objective as outlined above. For example, discrete electronic parts could be replæed with an application specific integrated circuitry (ASIC) and visa versa.
1 5 Definitions The following definitions are applicable to the present invention. ~th.om~tic~l operations are also explained when a~lupliately associated with the definition.
The term "fitting function" refers to a mathematical expression of the relationship between the day of the menstrual cycle (typically, day 1 through 28) and the 20 hormone con~entration that is Illea~,ured on that day. Although many fitting functions can be applied to the conc~ ion data, three especi~lly useful functions will be described which provide a high degree of certainty for the perfollllance of the present mventlon.
a. Linear Function. The first fitting function is a simple linear equation and 25 has the advantage of simplicity, fast c~lcul~tion time, and easy i~ ,le..,~"~ ion. The linear fitting function has the following formula:
concell~dtion = aO + (a1 x Day) where aO is the hltel-;ept of the linear fitting function, al is the slope of the linear fitting function and Day is the day of the IllensLl,lal cycle with Day=1 being the first day of ~l762~.8 menses. aO and a1 are numeric constants determined by the method of least squares.
The unit of concentration in this equation is that used in the collc~onding assay method. For example, nano-mole per liter is commonly used for assaying estrone 3-glucuronide (ElG) by immunoreaction. The linear least square plocedulG is easily 5 calculated and is well known or can be readily found in any number of reference texts.
e.g., "FORTRAN Programs For Scientists and Engineers", by Alan R. Miller (Sybex Inc., San Francisco (1982).
b. Linear Inverse Exponential Function. The second preferred function is a non-linear one and has the following formula:
concentration = ( aO + (al x Day))/ (I + l/lO(a2+ Day)) where a~, al and Day are as defined above, and a2 is another numeric consk~t determined by the method of least squares. This non-linear fitting function is more 15 complex since the cA~oncl.Lial CollllJoll."ll~ require more Culll~uLillg power and time to execute. However, with its complex nature, it provides an oppollullily to develop the adaptive algorithm to be ~ cucsed below. The llulllelic col.~lal1t~ of the function are estim~tr~l by the method of least squares to provide a -,;.,;"""" squared difference between the concclllldlion predicted by the fitting function and the actual concc--L,dlion 20 observed. For the non-linear fitting function, a2 is the exponential modulus. The use of this function is more complex and usually involves multi-variable o~l;,,,;7i~'ion iterative search. The Simplex method is one such well known method and is suitable for use in the method of the invention. A detailed rlc srnrtion of the Simplex method can be found in standard linear ~l.~l,.~lllll;l~g ~lobl_.ll solving texts such as "Mathematical 25 Progr~,u,.ir~g'', by Reinfeld and Vogel, Published by Prentice-Hall, Inc. Englewood Cliffs, N.J. (1958). Briefly, the Simplex method iflpnti~lps a basic feasible solution for the con~lldilled multi-variable equations. Then, the method is used to dcl~ ,--;-,e whether an (~)~illlUIII value has been reached by ~ub~l~clillg the value c~lcul~trd from the fitting model for each basis point from the actual analytical value at the point. This dirrclcllce Wo 95/16920 ~ 1 ~ 6 2 ~ 8 PCr/US94/14455 is squared and the squared difference is used to rank the sets of basis parameters. If an Op~illlUIII has not been reached, the method provides a way of moving to a new solution that will have a better value of the objective function. In the method of the invention, for example, four observed hormone concentrations on four consecutive days are set up as S four .cimlllt~nrous equations with a set of starting estim~tPd values of aO, al, and a2 with 4 slack variables, one for each of the four equations in a tableau. These variables are tested to see whether all the net gains are negative. If it is, then the OpLh~u~ has been found. If not, the operation of pivoting is carried out using the most marginal net gain variable as the pivot element and removing the variable with the least positive net l O gain to arrive at a new basic feasible solution with modified variables. The process is repeated until the o~Lilllulll has been found. The results are a set of values for the fitting function that give least square deviation for the observed hormone concentration.
c. Arit~unetic Average Function. The third function is a simple ~, ;LI ,., If ~liC
average c~lcul~tion. By using average values from a selected range of days as a baseline 15 to set up threshold values for comr~ricon to the observed value of a later day, a mathematical relationship is set up. This function is the most simple to use but not very reliable if used to predict beginning of the fertile cycle because it is more sensitive to data variability than the previously descrihecl two functions. However, using a sophisticated function for prediction of the be~ g of the fertile period to st~tictir~lly comren.c~te 20 for the data variation difficulties, such as the linear equation or non-linear equation as described above, a simple method could be used for predicting the end of the fertile period.
The term "slope" refers to the rate of change in hormone concentration. For the linear fitting function, it is equivalent to the value of a1. For the non-linear fitting 25 function, it is the evaluation of the first derivative of the fitting function.
The term "relative slope" refers to the value of slope mllltirlird by 100 then divided by the average values of the obse, ~d hormone conce"ll~Lion in the fitting function. In other words, relative slope is the average concentration norm~li7~d rate of p~"ce~L~ge concentration change. Since function fitting is done on the data in base ~1762~
- wo 95/16920 ~ ~ PCT/US94/14455 window, the slope is calculated from data in base window. [Define 'Base Window']The use of relative slope instead of slope itself reduces sensitivity of decision making to the individual to individual variation.
The term "predicted value" refers to the threshold value of hormone 5 conce~LIdLion, based on the fitted function, e~t~tiing one day beyond the fitting period.
For example, if a four day fitting window is used the predicted value of today would the evaluation of the fitted function with the variables least square fitted from the observed hormone concentration of days from 4 days ago to yesterday.
The term "threshold value" refers to the value of the sum of predicted value and10 the value of fitting standard deviation multiplied by a chosen multiplier. By co",l.~. ;llg the threshold value to the observed hormone concentration, the mathematical processes in the present invention de~e~ P whether a change in the trend in the hormone conce..l.d~ion has occurred. The multiplier is similar to a st~tictic~ 't' test and is selected using similar considerations in that the multiplier ~'~csessçs~ the confiflPn~e level to be given an ~csçs~,nr,.l that an observed change in hormonal concellllalion warrants an action. For the third fitting function of simple average as described above, the threshold value is the product of "baseline value" with a pre-set multiplier.
The term "data variability" refers to that property of the concentration ~e~u~ L~ that reflect a female's own day to day hormone concenLIdLion changes and result in minor deviations from a smooth slope. For example, variability can be caused by the changes in the way the hormone is metabolized in liver, the rate of hormone secretion into the kidney, the volume of liquid secreted by kidney, the conce"lld~illg process of urine in the urinary bladder, and variations in the assay method used to quantify the hormone conc~LI~tion.
The term "b~cPIinP" refers to the hormone concellL,dLions as nle~ulc;d on several col-~e~-uli-.re days that are characterized by relatively low values. Thel.,folc, the baseline values collc~L~ond to days where an increase of ovarian activity is unlikely. Typically, b~cPIinP for ElG l~,~lcse~ day 2 to day 5 of each cycle while the b~cPIinP for PDG
Ic~lcsellL~ the first 4 days of PDG measu,e,l,t;,ll of each cycle.
2 1 7 ~ 2 ~ & PCrlUS94/1445~ -The term "minim~l average value" refers to the minim~l value of a series of four day moving averages.
The term "fertile period" refers to that time of the menstrual cycle within which the presence of a viable sperm in the uterus, e.g, via sexual intGI~;oul~,e, would likely S leadtoconce~ ion.
The term "score" or "trigger point" refers to a number that is used to keep a record of an event. For example, the score is set to 0 when the menstrual cycle data analysis starts. When the observed assayed analyte concellL.dlion is higher fhan the threshold value and fulfills the decision rules as explained below, the score would be 10 incremented by 1.
The term "decision rules" refers to the set of logical relationships that define decision making in the algorithm. This set of rules includes, but is not limited to the following: the dirrGIGnce between the observed concentration collly~uGd to the expected concclllldlion; the relative slope colllpdlGd to the preset threshold of relative slope value;
15 the preset number of trigger points required for action colll~,d to the ~rCum~ tr(l trigger points of the cycle up to the day the decision is being made; and the maximal allowable fertile period. For example, a higher observed concelllld~ion than expected will give a trigger point if the slope is also positive. A trigger point will be given for each day having slope higher than threshold unless it is one of the baseline days. If the 20 number of trigger points accum--l~f~d is equal to or greater than the number of trigger required for action, the beginning or end of the fertile period is declared and the output means will accoldillgly take action by displaying information and the colll~uL~Iional and scoring means are prepared for the next phase, i.e., switching to accept an altern~tr assay result, e.g., PDG, if the beginning of the fertile period has been declared, or not 25 æcepting further assay results until the next cycle starts if the end of fertile period has been declared. For using the simple average function as Irsr-ribed in the "fitting function" section above, there is no relative slope consideration in the decision rules.
Instead, an additional rule is inrl~ d that takes into account the number of days from the baseline c ~lcul~tion. As an example for this particular case, from the day after the - WOgS/16920 ~1 7 6 2 ~ 8 PCT/US94/14455 pre-set number of days from beginning of the baseline data of PDG assay, an observed value higher than threshold value would give a trigger point.
Homone Mea~,ulcl.lcn~, As stated above, hormone Illcasulclllcll~, may be made following techniques well known in the art. The assay method can use any of a variety of established biochemical or immunological procedures that can be used in either liquid format or in dry Illclllbl~u~e bound solid phase format, such as chemical color reaction assay, enzyme-linked immunoassay (ELA), or radiolabeled immunoassay (RIA). For example,US Patent No. 4,138,278 describes a colorimetric immunoassay with enzyme label on solid surface for home pregnancy and ovulation tests, US Patent No. 5,120,643 describes a process for immunochlulllalography with colloidal particles, Japanese published application 1 1 09262-A describes an immunoassay of estrogen hormones in body fluid having two or more antibodies to estrogen (conjugate), estradiol (conjugate), and/or estratriol (conjugate) immobilized on a solid phase, and Japanese patent application 88052705-B described mP~cl-ring ~g~ ;"~lion of latex reagenl~, for diagnosis by measuring an increase in optical absGIl.auce. For ease of use and convenience a solid phase format is preferred. The numerous methods and devices suitable for use in a solid phase format are well known in the art and no special mention need be made in this respect.
The hormones or hormone metabolites to be assayed may be obtained from any biological fluid in which such collll.uullds are seclcted by the female. Typically, blood and urine are preferred, an early morning sample of urine being the most preferred fluid.
For in~t~n~e, when the hormone estrodiol is Illca~ulcd, the assay can be estrone or other estrogen hormone in blood or the metabolites of the estrone hormones in urine such as estrone 3-glucuronide (ElG). U.S. Patent No. 3,544,868 provides a suitable method which can be used to dclclll~ine steroid hormone glucuronide in urine samples byimmunoassay with tracers of glucuronide bound to enzyme. In a sirnilar manner, progc~,lcrone may be assayed using suitable markers such as serum progcslelone or a wo 95/16920 217 6 ~ 4 ~ PCT/US94/1445 progesterone metabolite in urine samples such as pregnanediol-glucuronide (PDG).Again, any number of methods are known to those skilled in the art, such as gas chro,l,atography, high pclrollll~lce liquid chromatography, enzymatic cletermin~tion, immuno reactions, etc. and are suitable for use to practice the present invention.
The means by which the daily ",ea~u,c",c"~ of hormone concentration is acco",~ I,Pd are not i",~u,~ll to the present invention. Any suitable means can be used including visual observation, spectrophotometric reading, radioactivity counting, or other electrom~gn~o-tic readings as applu~,iate for the chosen assay method. VVhen a solid phase format is used the me~su,G,,,cnl can be made most typically by a reader which detects and records results of electromagnetic re~Aingc. Readers app,o~,iate for the chosen assay methods are well known and the above mentioned references illustrate the wide range of read out formats available. In a similar manner, the data storage means, co",~u~lional capacity, evaluation means, scoring means, and output means can use any number of well known methods for acco",~lishing these functions. However, it is preferred to illCOl~OI~-t, the above means into a single hl~llu",c"l that can function as a unitary device that would require esse~ti~lly no input from the user other than to input a daily assay to be measured.
All ~cfc~el~ces to patents or publications in this specifi~tion are inc~ JIdled herein by Icrclcnce.
Examples Example 1. Detection of the beginning of fertile period In this example, the begill~ g of fertile period is predicted using a linear fitfunction and decision rules having the following p~l,c~ observed hormone c~ e~ alion greater than 2 times minim~l average value for trigger point con~ Pration, 6% relative slope is set as threshold after day 5, threshold value is set by adding 5 times standard deviation to the predicted value, and a score of 4 are required to declare action.
As would be understood, this set of parameters is one of the many sets which may be used to assess the performance of the method of invention on a large data base. The particular p~alllc~ used here show the application of the method of the invention and _- WO g5/16920 ~ 1 ~ 6 2 4 8 PCr/US94/14455 the resulting information which is provided by such data analysis. The o~ ulll set of p~alllct~l~ for a large data base can be found by pclr~,lllli,~g a ~çsigned experiment of systç~n~ti~ analysis using known ovulation day for reference verification. Further details of the o~i"liz~lion of parameters is found in Example 3 described below.In this example the estrogen hormone estrone 3-glucuronide (ElG) is measured in urine. The plucedulc utilizes the World Health Olga~ alion radioillullul,o assay (RIA) procedure. The first ElG assay is done on day 2 instead of day 1 so that a clear definition of the start of cycle is given and a simple daily routine is set up to fæilitate sample collection and assay pelr~" " ,~ e The sample is collected from the first early morning passage of urine. Hormone assay is obtained for at least four days with results from eæh day entered into a lll-c~ol.locessor pl-)~lalllll,ed for linear function calculation The observed data of the first five days of the cycle are shown in Table 1.
The observations of Days 2-5 are fitted using the linear function and the various values described above are derived. In this case, the relative slope is not taken into account to increment the score because it is not after day 5 as stated in decision rules that relative slope fætor would be considered only after day 5. Therefore, no adjustment is made to the score. The minim~l average value is the same as average value observed because this four day period is the first window observed. Further, there is no predicted value c~lcul~tion and the threshold value is set to a high number to indicate that no comparison is needed. On Day 6 the daily hormone lllea~ulclllclll is made. The results are shown in Table 2.
The working window is shifted to day 2 to day 6 with a base window of day 2 to day 5. The observed day 6 value (89.35) is then analyzed according to the method of the invention. In this case the slope is positive (20.28) but the observed hormone assay value (89.35) is not higher than twice the minim~l average value (76.35x(2)=153.7) and thus there is no change in the score. However, since the relative slope observed is higher than threshold slope (6) and the day is later than day 5 the score is inclclllcnled by 1. The score which results is less than set number of 3 and, the,efolc, day 6 is not declared as the beginning of the fertile period. The process of day 6 is repeated on day Wo 95/16920 21~ 6 2 4 8 PCT/U$94/14455 7. The window shifts to Day 3-7 with a base window of day 3 to day 6. The data observed for day 7 and the proces.cingc are shown in the Table 3.
Since the observed value (120.37) is not higher than 2 times minim~l average value (153.7), no score is inclclllclllcd . Again, since relative slope (15) is higher than 5 set threshold (6), the score is il~crc~cnted by 1. The resulting score is not higher than that required for action and therefore Day 7 is not declared the beginning of the fertile period. This process is repeated until an action point is reached and the beginning of fertile period is declared. Table 4 shows the p~ucesshlg of data from other days.
On day 10, the score reaches the required number for action, namely 4. Thus 10 the beginning of the fertile period for this cycle is declared on day 10. The ovulation day for this particular cycle had been in~epen~ently established as day 13 by the daily ultrasound obstl~ation, i.e., day 13 showed characteristic reduction in follicle diameter as well as other features intlir~ting ovulation such as echo observed in the follicle sac and blurring of follicle borders, etc. Therefore, in this example the method of the 15 invention declares the beginning of fertile period 3 days prior to ovulation.
Example 2. D~t~ - . . .; .-~l ion of the Ending of Fertile Period This example describes the method of the invention for predicting the end of the fertile period. This example uses the same menstrual cycle as the one used in Example l 20 and measures urinary pregnanediol 3-glucuronide (PDG) assay by the World Health O,ganization radioi--..-.~..o~cs~y method described above.
Example 2(a) This example uses the linear fitting function with the following p~U~ull~l..; 1 25 times minim~l average value, 5 times standard deviation for threshold value, 12%
relative slope, and a score of 3 for declaration of end of fertile period. Table 5 shows the observed concc;"~ion values and other parameters as analyzed for days 11 through 16 accordi~g to the method of the invention.
A four day baseline data is collected for initial relative slope and threshold value wo 95/16920 ~ ~ 7 6 2 ~ 8 PCr/US94/1445S
calculation similar to the procedures described above for Example l. In Example 2(a), day 11 to day 14 provides the four day baseline data of PDG. On day 15, PDG
concentration observed, 11.54 uM, is higher than 1 times minim~l average value and higher than threshold value of the day. Thus, the score is in~ lented by 1. The score is further h~ulcl~el~led by 1 due to a relative slope higher (29.99) than threshold (12) in the base window. However, the ~cunmll~t-ocl score of 2 remains less than that required number for action and, therefore, this day is passed without declaring the end of fertile period. On day 16, observed value is greater than threshold value and score is thus incremented by 1. The resulting i~l~;lc;lllen~al score (3) is the number preset as required for action and day 16 is declared as the end of the fertile period for this particular cycle.
This is three days after ovulation as d~ ...;..P-d by ultrasound (Example 1). Noadditional hormone assays are needed until the be~,h,..i..g of next cycle. At the first day of menses of the next cycle, the procedure for predicting the beginning of fertile period is carried out as ~l~s~ ~ibe~l in Example 1.
Example 2(b) This example uses the third fitting function of ~i ;LII~ ir averaging function.
The pal~lletel~ used are: 5 days after declaration of beginning of fertile period to begin fitting function for ending of fertile pe;iod, a value of 2 times the base value as the threshold value for ending of fertile period, and a score of l to declare action. In this case the multiplier for standard deviation is zero. The baseline data is obtained from day 11 to day 14. The ~ ",~lir average method c~lrlll~te~l the b~cP-line value to 6.5 llM.
The threshold value is then set to 13.0 ~lM (2 times base value; as set in the p~lletel~).
As shown in the Table 5, day 15 showed 11.54 ~M which is lower than the threshold value of 13Ø Therefore day 15 is passed without action. On day 16, the observed PDG value is 13.28 ,uM, i.e., higher than threshold value. The score is in-;lel..~ d to l and the ending of the fertile period is declared because a score of 1 is pre-set for decl~aLion of action. Again, the ending of the fertile period is three days post-ovulation as l.le~ul~d by ultrasound (Example 1).
wo 95/16920 - ~ PCr/uS94/1445 Example 3. Finding the best set of pal~llllGtGl~i for a large data base By collecting and analyzing a large d~t~h~ce co~ lg ~ e.u-ls data points, one can C~/tillli~,G the values ~cign~d to the pre-set pal~llGlGI~, e.g., minim~l average 5 value, relative slope, threshold slope, multiplier, and the score. In this way the method of the invention can be cnctomi7~d to adjust for earlier detection and declaration of beginning of fertile period (for cû--l~a~G~ e utility) or to adjust for later (i.e., closer to ovulation) detection and declaration of beginning of fertile period (for concGp~ion enh~ncement utility).
For this purpose, a large data base with known ovulation day is required. A
collllllonly practiced expG.il--e-,~l design can be used for the ol"i--,i;~alion of ~aldlllclGI~.
Briefly, it involves many G~. h l lr~ runs with regularly varying combination ofpalalllG~Gl~i. The trends of the effect of varying p~alllet~ are analyzed according to the desired relationship to the IGÇ~,lGnce ovulation day. This analysis is then used to predic the best set of parameters for the desired relationship.
Tables 6-8 shows the results from a set of data base collected from 170 cycles.
The time of ovulation of each menstrual cycle is t~ rrninPd by ultrasound investigation to within 24 hours CGI lai--ly. Ovulation is declared when the sonographer notes both a reduction in follicle size and a change in follicle apl)e~dllce by echo pattern and follicle border after rupture. The urinary ElG and urinary PDG assays are done following the RIA method of Examples 1 and 2. The pdlalllGtGI~ tested for this run on this data set are as follows:
For beginning of fertile period:
a minim~l average value of 2.25 times background, 4.5% relative slope, a multiplier of 3.5 times sl~d~d deviation, and a score of 4 for declaration of action.
For end of fertile period:
6 days after declaration of be~,i--l i--g of fertile period to begin fitting function, 2 times minim~l background value, 4.5 times background as threshold - Wo 95/16920 2 1 7 ~ 2 ~ 8 PCT/US94/14455 value, a score of 1, and 11 days as maximal allowable fertile period.
Table 6 shows the results of predicting the be~ g of fertile period of 170 cycles. The first colurnn of the table lists the categories of results relating to ovulation day. For example -9 I~ Stll~ a result that beghlnil,g of fertile period is declared 9 S days before ovulation occurred. The second column pleselll~ the number of cycles that are predicted to have the beginning of fertile period as the co,~ onding number of days from ovulation in the first colurnn. The third column presents the number in the second column as a per~el-~ge fraction of the total number of 170 cycles. The fourth column .~..,sc..~ the cnmlll~t~d fraction of the cycles that have been predicted to have begun fertile period before and on the day shown in the first column before ovulation. The last colurnn represents the 1~ fraction of cycles that have not been announced to have entered fertile period. As can be seen in Table 6, a high majority of cycles (72%) are declared to have begun fertile period 3 days before ovulation and only a small fraction (5%) had not been declared to have entered fertile period by 1 day before ovulation.
Similarly, Table 7 shows the results of testing for the ending of fertile period of 170 cycles. The column and rows of data are arranged and defined as shown in Table 6. As can be seen in the cumlll~tPd fraction column, less than 5% of cycles are announced to have ended fertile period on and before the day of ovulation (as det~ ,I-;--~d by ultrasound), while a majority of cycles (55% to 73%) are announced to have ended the fertile period by the time 4 to 5 days after ovulation.
As can be seen from these results, the predictive value of the method of the invention is superior to other methods known in the art.
Example 4. Comparison to known methods.
As an additional example, the results of the present invention are col,lpd-ed to those published by J.P. Royston, Statistics in Medicine, 10:221-240 (1991). The Royston reference ~ullull~i~ed and ~csPssecl the ~l~llll~lce of various predictive tests of fertile period based on published data. The data are incol~ul~ed into Table 8 and exp~n~l to include data generated by the method of the invention. As is known in the 2176~
wo 95/16920 ` ` PCr/US94/1445 art (see, e.g., pp. 231 of Royston) a commonly accepted criteria for a critical fertility period inclu-les the period of three days pre-ovulation and two days post-ovulation. In Table 8 p~culæltl g is defined as the fraction of cycles that provided a fertile period that overlaps the critical period. P~læter T is an e~ e of the dispersion of the 5 distribution of the fertile period around the ideal value of 6. It is c~l~ulrtç~l by using the following formula:
T = ~ ( f a - 6) 2 + sf 2 } 0-5 10 where fa stands for the average value of the fertile period and Sf stands for the standard deviation of the fertile period.
In Table 8, data from dirr~ sources are colll~ d in terms of the average ~bstin~n~e days and the standard deviation of the sarnples in their respective study in practicing each method. The g p&~ll~,t._l c~ lr~ed above using data ~ t~,d 15 following the method of the invention shows lower values than prior art methods for the reason that all the available data in the sample cycles were used in the c~lculrtion. For a practical use of the method, it would be more a~,u~iate to include those cycles that did not give a~prop,iate signals. Thus, when the cycles classified as not giving a~,op,iate signals are also included into the crlcul~tion, the g values would about 0.61 to 0.64 for 20 the chemical test l and ch~o-rnic~l test 2 in Royston's table. If such g values are used, the values co,l~al~ well with the results ge~ led using the method of the invention (0.61).
The smaller T value which results when using the method of the invention in-lic~t~s that the present invention is capable of giving a much better defined fertile period in a more reproducible manner. Thus, one advantage of the present invention over the prior art 25 methods of predicting fertile period is that a much shorter fertile period is ~sign~1 to each cycle.
Example 5. Trading o~ lion for a shorter predicted fertile period.
Examples 3 and 4 show a set of optimal p~u~net~ which provide good - WO 95/16920 ~ ~ 7 6 2 ~ 8 PCT/US94/14455 p~lro~ ce and an acceptable predicted fertile period. This example shows that the palalllc;~ can be dt;l~ll~ined by following the expelilllt;lllal design used above but with the goal of a shortened predicted fertile period. This provides an additional advantage of fewer testing days which reduces the number of tests which must be taken and provides S a more cost effective and easier to use method.
The o~Lillli,~lion uses a 169 cycle data base from the same participants as used in Example 3, however, one of the 170 cycle data base was ~icqu~lified due to consecutive missed samples half way through the cycle. Ovulation of each menstrual cycle is detelmined by ultrasound investigation to within 24 hours cel laillty as before.However, in this case, the first urine sample collected for analysis is done on day 6 of menses. The urinary E 1 G and urinary PDG assays are done following the RIA method described above. The palalllc;l~l~ are set as follows:
For beginning of fertile period:
Analysis begins on day 6 of menses, no minim~l average value required above background, 4.5% relative slope, a multiplier of 3.5 times standard deviation, and a score of 3.
For end of fertile period:
No minim~l average value required above background, 5% relative slope, a multiplier of 4 times standard deviation, a score of 4, and 10 days as maximal allowed fertile period.
Table 9 shows the results of predicting the beginning of fertile period of these169 cycles. As can be seen in Table 9, a majority of cycles (59%) are declared to have begun fertile period 3 days before actual ovulation (as verified by ultrasound) and only a small fraction of cycles (3.6%) had not been declared to have entered fertile period by the day of ovulation.
Table 10 shows the results of testing for the ending of fertile period of the 169 cycles. As can be seen in the cum~ t~od fraction column, less than 4% of cycles are predicted by the method to have ended fertile period on or before the day of actual ovulation (as ~Ir.l~ "~ Pd by ultrasound), while a majority of cycles (48% to 67%) are Wo 95/16920 PCT/US94/1445~
217 62~8 26 predicted to have ended the fertile period by 4 to 5 days after actual ovulation.
A comparison of the results from Example 3 (the method of the invention commencing on day 2 of menses) and Example 5 (day 6) is shown in Table 11. As can be seen, the pararneters accign~d in Example 5 provide certain advantages over those 5 used in Example 3. The predicted length of fertile phase is reduced from 8.4 to 8.1 days with a conco,~ reduction in standard deviation of fertile phase from 2 to 1.5.
However, this reduction in predicted fertile period results in a reduced index (g) of performance.
The effect of this reduction in g value on the predictive value of the method may 10 be less than the nllm~-rir~l value would indicate since it appears that the reduction is predominately due to a difference of three vs two days pre-ovulation in predicting the start of fertile period. Furthermore, the lower g value may also be offset in part on the presumed use of an early morning urine sample. Therefore, the slightly lower g values which result from col.llll~ncillg the method of the invention on day 6 rather than day 2 15 are well co~ e~ ed by the hlcleased benefit of a shorter predicted fertile period and fewer testing days.
Example 6. Detection of the beginning of fertile period In this example, the begillllillg of fertile period is predicted using a linear fit 20 function and decision rules having the following p~al~ 3% relative slope is set as threshold after day 5,expected value is set by adding 3 times standard deviation to the predicted value, and 3 trigger points are required to declare action. As would be understood, this set of p~lle~ is one of the many sets which may be used to atssess the pelr(,llllà,lce of the method of invention on a large data base. The particular 25 p~alllelel~ used here show the application of the method of the invention and the resulting hlfc,lllldlion which is provided by such data analysis. The l lhl~ulll set of p~al"etel~ for a large data base can be found by pe,rolllli"g a ~lesigned exlJ~lilllelll of system~tiC analysis using known ovulation day for reference v~fir,caLion. Further details of the o~ .d~ion of pal~llel~ is found in Example 8 described below.
zl76248 - wo 95l16920 PCr/US94/14455 In this example the estrogen hormone estrone 3-glucuronide (ElG) is measured in urine. The procedure utilizes a solid phase immllno~c~y on nitrocellulose. In this example in contrast to Example 1, the first ElG assay is done on day 5 instead of day 1 from the start of menses. The advantage of this most p,Grclled method of the invention is that it reduces the number of assays per cycle required to be performed by the user as well as reducing the possibility of urine samples being co~ ted by ~,ellsllual flow.
Again, the sample is plcrcl~bly collected from the first early morning passage of urine.
Samples and assays are obtained and completed for the first four days (i.e., day 5 to day 8 of the menstrual cycle) with assay results from each day read and stored in the meter.
The meter are programmed with the mathematical function of this invention. Thesefirst four days assay results serve as data the baseline data as explained above. As explained above, in the preferred method the meter is programed with function of the invention and thus the actual results are not viewed by the user. Accordingly, the internal operation of the c~l~ ul~tion process inside the meter is ~lesçnhed here in order to more fully explain the method of the invention. The observed data of a first four days results are shown in Table 12.
The observations of Days 5-8 are f tted using the linear function and the other related values described above are derived. Under the pal~llcL,~ set out above, the first four days serve as the baseline days, and, therefore, no trigger count adjustment is made. This results in the minim~l average value being the same as the average value observed because this four day period is the first window observed. Furthermore, there is no predicted value c~lrul~tion and the expected value is not applicable. On Day 9 a daily hormone llle~ulcllltllt is made, the results of which are shown in Table 13.
The working window is now shifted to day 5 to day 9 with abase window of day 5 to day 8. The observed day 9 value (30) is then analyzed ~ccording to the method of the invention. In this case the slope is positive (14) but the observed hollllone assay value (30) is not higher than the minim~l expected value (67.9) and thus there is no trigger point for this feature. However, since the relative slope observed (14) is higher than threshold slope (3) and the day is later than day 8, the trigger point is inclGlllcntcd Wo 95/16920 ~ 1 7 6 2 ~ 8 PCT/US94/14455 by 1. However, the trigger point reached ( 1) is less than set number of 3 and, therefore, day 9 is not the beginning of the fertile period. The process of day 9 is repeated on day 10. The window then shifts to Day 6-10 with a base window of day 6 to day 9. The data observed for day 10 and the results of the method of the invention 5 are shown in the Table 14.
In Table 14 it can be seen that the day 10 observed value (23) is not higher than the expected minim~l average value (66.9), and this is no trigger points are il~;lclllcll~cd by this criterion. Again, since relative slope (17.6) is higher than set threshold (3), the trigger point is hlclclllcllled by 1. The resulting trigger point reached (2) is not higher 10 than trigger number required for action and therefore Day 10 is not declared the beginning of the fertile period. This process is repeated until an action point is reached and the beginning of fertile period is declared. Table 15 shows the proces.cing of data on day 13 and the data from day from 9 to day 13 are displayed.
On day 13, the trigger point reaches the required number for action, namely 3.
15 Thus the beginning of the fertile period for this cycle is declared on day 13. The ovulation day for this particular cycle had been in-lç.pçnr1~ tly established as day 17 by the daily ultrasound observation, i.e., day 17 showed cll~a~;lclistic reduction in follicle di~mr-tçr as well as other features in-lir~ting ovulation such as echo observed in the follicle sac and blurring of follicle borders, etc. Therefore, in this example the method 20 of the invention declares the begilln;llg of fertile period 4 days prior to ovulation.
Example 7. Detç~ ",;n,.lion of the Ending of Fertile Period This example describes the method of the invention for predicting the end of the fertile period and uses the same l"~,nsl,ual cycle as the one used in Example 6. Urinary 25 estrone 3 glucuronide (ElG) is measured by the solid phase immunoassay as desc,ibcd above in Example 6. After the begi""ing of fertile period is decl~ in FY~mple 6 day 13, the daily assay of urinary ElG continues until the pre-set day of day 14. On that day c~lr,ul~tions co,..",~nre using the same running window scheme of five days as that used for pleliclillg the begilmillg of fertile period in Example 6, except the relative - WO95/16920 ~1 762~8 PCrlUS94/1445 slope is c~lcul~t~d from the day of observation itself. In this example the following decision rules are used: default fertile period is 9 days long, reduce fertile period by one day if maximum relative slope is greater than set threshold of 25%, extend fertile period by one day if the beginning of fertile period is less than 10 days while maximum relative S slope is negative, reduce fertile period by 2 days if m~imllm relative slope reached was between day 11 and day 12, reduce fertile period by 1 day if the maximum relative slope is on day 13. Table 16 shows the observed concentration values and other parameters as analyzed for day 9 through 14 according to the method of the invention. As shown in Table 16, the relative slope (42.4%) observed on day 13, is greater than the relative slope of day 14 (13.0). According to the decision rules, a one day reduction infertile period is given for a relative slope greater than threshold (25%) and a fertile period is reduced by one day for reaching maximum on day 13. Therefore, the fertile period for this cycle is 7 days and the end of the fertile period for this cycle is calculated to be day 20. Ovulation day for this menstrual cycle had been detc.lllilled by the ultrasound 15 reference method on day 17 as mentioned above in Example 6. Thus, this declaration of end of fertile period provides 3 additional subsequent days to ovulation for contraception purposes.
Example 8. Determination of Optional P~alllclcl~
By collecting and analyzing a large d~t~h~ce C~ .;Ui.. g llul~crous data points, one can o~ e the values ~c~ign.od to the pre-set pal~llclcl~, e.g., minim~l average value, relative slope, threshold slope, multiplier, and the trigger number. In this way the method of the invention can be cu~ Jllfi~cd to adjust for earlier detection and declaration of beginning of fertile period (for col,ll~cc~ e utility) or to adjust for later 25 (i.e., closer to ovulation) detection and declaration of begi"nillg of fertile period (for conception cl~h~ 'c~e~l utility).
For this purpose, a large data base with known ovulation day is required. A
cornmonly practiced eA~e. ;lll~ design can be used for the u~hl~izalion of p~halllGlcl~..
Briefly, it involves many eA~- - ;",~ runs with regularly varying combination of Wo 95/16920 2 1 7 6 ~ 4 i~ PCr/US9411445~
parameters. The trends of the effect of varying pa,~ne~cl~ are analyzed according to the desired relationship to the reference ovulation day. This analysis is then used to predict the best set of parameters for the desired relationship.
Tables 17-19 shows the results from a set of data base collected from 120 cycles. The time of ovulation of each menstrual cycle is dete- ,II;II~d by ultrasound investigation to within 24 hours certainty. Ovulation is declared when the sonographer notes both a reduction in follicle size and a change in follicle a~pe~dllce by echo pattern and follicle border after rupture. The urinary ElG assay çc.centi~lly follows the solid phase immunoassay as tlPs~-rihecl in Examples 6 and 7. The p~ulle~.s tested for this Example on this data set are as the same as described in Example 6 and 7.
Table 17 shows the results of predicting the beginning of fertile period for 120cycles. The first column of the table lists the results obtained by the method of the invention as cOlll~alcd to actual ovulation day. For example 4 ,c~lcsenL~ a result that the present method declares the beginning of fertile period 4 days before a confined ovulation had occurred. The second column plcs~.lL~ the number of cycles that are predicted to have the beginning of fertile period as the coll~s~ondillg number of days from ovulation in the first column. The third column plcScllL~ the number in the second column as a pel-;clll~ge fraction of the total number of 120 cycles. The fourth column Ic~lcScllL~ the cum~ tecl fraction of the cycles that have been predicted to have begun fertile period before and on the day shown in the first column beforc ovulation. The last column lc~lcscll~s the lClll~ g fraction of cycles that have not been announced to have entered fertile period. As can be seen in Table 17, a high majority of cycles (66%) are declared to have begun fertile period 3 days before ovulation and only a small fraction (8%) had not been declared to have entered fertile period by I day before ovulation.
Similarly, Table 18 shows the results of testing for the ending of fertile period of 170 cycles. The data is arranged and defined as in Table 17. As can be seen in the cum~ ted fraction column, less than 8% of cycles are announced to have ended fertile period on and before the day of ovulation (as detel"l",ed by ultrasound), while a majority of cycles (43% to 59%) are announced to have ended the fertile period by the 2~76248 --- WO 9S/16920 Pcr/uss4/l4455 time 4 to 5 days after ovulation.
As can be seen from these results, the predictive value of the method of the invention is superior to other methods known in the art.
5 Example 9. Co~ alisol1 to known methods.
As a further collfi,llla~ion of the superior predictive use of the present invention, the results of the present invention are compared to those published by J.P. Royston, Statistics in Medicine, 10:221-240 (1991). The Royston reference ~.ulll~l~izes and assesses the performance of various predictive tests of fertile period based on published lO data. The data are incorporated into Table 19 and expanded to include data generated by the method of the invention. As is known in the art (see, e.g., pp. 231 of Royston) a commonly accepted criteria for a critical fertility period includes the period of three days pre-ovulation and two days post-ovulation. In Table 19 parameter g is defined as the fraction of cycles that provided a fertile period that overlaps the critical period.
15 Parameter T is an çstim~ts of the dispersion of the distribution of the fertile period around the ideal value of 6. It is calc~ tP-d by using the following formula:
T={(f - 6)2+ sf 2}0.5 20 where fa stands for the average value of the fertile period and Sf stands for the standard deviation of the fertile period.
In Table 19, data from dirr~.Gnt sources are colllp~;d in terms of the average abstinence days and the standard deviation of the Salllpl-~ in theimcs~ ;Li~e study in practicing each method. The g pdl.:ullt;Lcl c~lrlll~tecl above using data generated 25 following the method of the invention shows lower values than prior art methods for the reason that all the available data in the sample cycles were used in the c~lr,~l~ltion. For a practical use of the method, it would be more a~ ,iate to include those cycles that did not give ap~"o~liate signals. Thus, when the cycles cl~cci~lec~ as not giving a~pl~,~liate signals are also included into the s~lrlll~tion7 the g values would approximately 0.65 to Wo 95/16920 ;~ 17 6 2 4 ~` PCT/US94/14455 0.68 for the either chemical test l or chemical test 2. If such g values are used, the values co~ well with the results generated using the method of the present invention (0.68). The smaller T value which results when using the method of the inventioninrlir~tes that the present invention is capable of giving a much better defined fertile 5 period in a more reproducible manner. Thus, one advantage of the present invention over the prior art methods of predicting fertile period is that a much shorter fertile period is assigned to each cycle.
As can be seen from these results, the predictive value of the method of the invention is superior to other methods known in the art.
Examples 8 and 9 show a set of optimal parameters which provide good performance and an acceptable predicted fertile period. As is understood by those skilled in the art, the parameters can be ~letermin~od by following the ex~.i",~ l design used above but with the goal of a shortened predicted fertile period. By applying the principle of.design of e~ , another set of ~alal~le~ could be devised to 15 provide an additional advantage of fewer testing days which reduces the number of tests which must be taken and provides a more cost effective and easier to use method. On the other hand, another set of parameters, similar to Examples 8, could be devised to provide higher protection with the understood expense in running more assays for a longer period.
The alternative emborlimrnt~ described are intrn~ d as examples rather than as limits. Thus, the description of the invention is not intrn-lPd to limit the invention to the particular embo-l;",~ ; disclosed, but it is inten-led to enco",~ass all equivalents and subject matter within the spirit and scope of the invention as observed above and as set forth in the following claims.
WO 95116920 2 1 7 6 2 ~ 8 PCr/US94/14455 TABLE 1- Day 5 Cycle Assay Relative Standard Minimal Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 2 62.61
The term "fertile period" refers to that time of the menstrual cycle within which the presence of a viable sperm in the uterus, e.g, via sexual intGI~;oul~,e, would likely S leadtoconce~ ion.
The term "score" or "trigger point" refers to a number that is used to keep a record of an event. For example, the score is set to 0 when the menstrual cycle data analysis starts. When the observed assayed analyte concellL.dlion is higher fhan the threshold value and fulfills the decision rules as explained below, the score would be 10 incremented by 1.
The term "decision rules" refers to the set of logical relationships that define decision making in the algorithm. This set of rules includes, but is not limited to the following: the dirrGIGnce between the observed concentration collly~uGd to the expected concclllldlion; the relative slope colllpdlGd to the preset threshold of relative slope value;
15 the preset number of trigger points required for action colll~,d to the ~rCum~ tr(l trigger points of the cycle up to the day the decision is being made; and the maximal allowable fertile period. For example, a higher observed concelllld~ion than expected will give a trigger point if the slope is also positive. A trigger point will be given for each day having slope higher than threshold unless it is one of the baseline days. If the 20 number of trigger points accum--l~f~d is equal to or greater than the number of trigger required for action, the beginning or end of the fertile period is declared and the output means will accoldillgly take action by displaying information and the colll~uL~Iional and scoring means are prepared for the next phase, i.e., switching to accept an altern~tr assay result, e.g., PDG, if the beginning of the fertile period has been declared, or not 25 æcepting further assay results until the next cycle starts if the end of fertile period has been declared. For using the simple average function as Irsr-ribed in the "fitting function" section above, there is no relative slope consideration in the decision rules.
Instead, an additional rule is inrl~ d that takes into account the number of days from the baseline c ~lcul~tion. As an example for this particular case, from the day after the - WOgS/16920 ~1 7 6 2 ~ 8 PCT/US94/14455 pre-set number of days from beginning of the baseline data of PDG assay, an observed value higher than threshold value would give a trigger point.
Homone Mea~,ulcl.lcn~, As stated above, hormone Illcasulclllcll~, may be made following techniques well known in the art. The assay method can use any of a variety of established biochemical or immunological procedures that can be used in either liquid format or in dry Illclllbl~u~e bound solid phase format, such as chemical color reaction assay, enzyme-linked immunoassay (ELA), or radiolabeled immunoassay (RIA). For example,US Patent No. 4,138,278 describes a colorimetric immunoassay with enzyme label on solid surface for home pregnancy and ovulation tests, US Patent No. 5,120,643 describes a process for immunochlulllalography with colloidal particles, Japanese published application 1 1 09262-A describes an immunoassay of estrogen hormones in body fluid having two or more antibodies to estrogen (conjugate), estradiol (conjugate), and/or estratriol (conjugate) immobilized on a solid phase, and Japanese patent application 88052705-B described mP~cl-ring ~g~ ;"~lion of latex reagenl~, for diagnosis by measuring an increase in optical absGIl.auce. For ease of use and convenience a solid phase format is preferred. The numerous methods and devices suitable for use in a solid phase format are well known in the art and no special mention need be made in this respect.
The hormones or hormone metabolites to be assayed may be obtained from any biological fluid in which such collll.uullds are seclcted by the female. Typically, blood and urine are preferred, an early morning sample of urine being the most preferred fluid.
For in~t~n~e, when the hormone estrodiol is Illca~ulcd, the assay can be estrone or other estrogen hormone in blood or the metabolites of the estrone hormones in urine such as estrone 3-glucuronide (ElG). U.S. Patent No. 3,544,868 provides a suitable method which can be used to dclclll~ine steroid hormone glucuronide in urine samples byimmunoassay with tracers of glucuronide bound to enzyme. In a sirnilar manner, progc~,lcrone may be assayed using suitable markers such as serum progcslelone or a wo 95/16920 217 6 ~ 4 ~ PCT/US94/1445 progesterone metabolite in urine samples such as pregnanediol-glucuronide (PDG).Again, any number of methods are known to those skilled in the art, such as gas chro,l,atography, high pclrollll~lce liquid chromatography, enzymatic cletermin~tion, immuno reactions, etc. and are suitable for use to practice the present invention.
The means by which the daily ",ea~u,c",c"~ of hormone concentration is acco",~ I,Pd are not i",~u,~ll to the present invention. Any suitable means can be used including visual observation, spectrophotometric reading, radioactivity counting, or other electrom~gn~o-tic readings as applu~,iate for the chosen assay method. VVhen a solid phase format is used the me~su,G,,,cnl can be made most typically by a reader which detects and records results of electromagnetic re~Aingc. Readers app,o~,iate for the chosen assay methods are well known and the above mentioned references illustrate the wide range of read out formats available. In a similar manner, the data storage means, co",~u~lional capacity, evaluation means, scoring means, and output means can use any number of well known methods for acco",~lishing these functions. However, it is preferred to illCOl~OI~-t, the above means into a single hl~llu",c"l that can function as a unitary device that would require esse~ti~lly no input from the user other than to input a daily assay to be measured.
All ~cfc~el~ces to patents or publications in this specifi~tion are inc~ JIdled herein by Icrclcnce.
Examples Example 1. Detection of the beginning of fertile period In this example, the begill~ g of fertile period is predicted using a linear fitfunction and decision rules having the following p~l,c~ observed hormone c~ e~ alion greater than 2 times minim~l average value for trigger point con~ Pration, 6% relative slope is set as threshold after day 5, threshold value is set by adding 5 times standard deviation to the predicted value, and a score of 4 are required to declare action.
As would be understood, this set of parameters is one of the many sets which may be used to assess the performance of the method of invention on a large data base. The particular p~alllc~ used here show the application of the method of the invention and _- WO g5/16920 ~ 1 ~ 6 2 4 8 PCr/US94/14455 the resulting information which is provided by such data analysis. The o~ ulll set of p~alllct~l~ for a large data base can be found by pclr~,lllli,~g a ~çsigned experiment of systç~n~ti~ analysis using known ovulation day for reference verification. Further details of the o~i"liz~lion of parameters is found in Example 3 described below.In this example the estrogen hormone estrone 3-glucuronide (ElG) is measured in urine. The plucedulc utilizes the World Health Olga~ alion radioillullul,o assay (RIA) procedure. The first ElG assay is done on day 2 instead of day 1 so that a clear definition of the start of cycle is given and a simple daily routine is set up to fæilitate sample collection and assay pelr~" " ,~ e The sample is collected from the first early morning passage of urine. Hormone assay is obtained for at least four days with results from eæh day entered into a lll-c~ol.locessor pl-)~lalllll,ed for linear function calculation The observed data of the first five days of the cycle are shown in Table 1.
The observations of Days 2-5 are fitted using the linear function and the various values described above are derived. In this case, the relative slope is not taken into account to increment the score because it is not after day 5 as stated in decision rules that relative slope fætor would be considered only after day 5. Therefore, no adjustment is made to the score. The minim~l average value is the same as average value observed because this four day period is the first window observed. Further, there is no predicted value c~lcul~tion and the threshold value is set to a high number to indicate that no comparison is needed. On Day 6 the daily hormone lllea~ulclllclll is made. The results are shown in Table 2.
The working window is shifted to day 2 to day 6 with a base window of day 2 to day 5. The observed day 6 value (89.35) is then analyzed according to the method of the invention. In this case the slope is positive (20.28) but the observed hormone assay value (89.35) is not higher than twice the minim~l average value (76.35x(2)=153.7) and thus there is no change in the score. However, since the relative slope observed is higher than threshold slope (6) and the day is later than day 5 the score is inclclllcnled by 1. The score which results is less than set number of 3 and, the,efolc, day 6 is not declared as the beginning of the fertile period. The process of day 6 is repeated on day Wo 95/16920 21~ 6 2 4 8 PCT/U$94/14455 7. The window shifts to Day 3-7 with a base window of day 3 to day 6. The data observed for day 7 and the proces.cingc are shown in the Table 3.
Since the observed value (120.37) is not higher than 2 times minim~l average value (153.7), no score is inclclllclllcd . Again, since relative slope (15) is higher than 5 set threshold (6), the score is il~crc~cnted by 1. The resulting score is not higher than that required for action and therefore Day 7 is not declared the beginning of the fertile period. This process is repeated until an action point is reached and the beginning of fertile period is declared. Table 4 shows the p~ucesshlg of data from other days.
On day 10, the score reaches the required number for action, namely 4. Thus 10 the beginning of the fertile period for this cycle is declared on day 10. The ovulation day for this particular cycle had been in~epen~ently established as day 13 by the daily ultrasound obstl~ation, i.e., day 13 showed characteristic reduction in follicle diameter as well as other features intlir~ting ovulation such as echo observed in the follicle sac and blurring of follicle borders, etc. Therefore, in this example the method of the 15 invention declares the beginning of fertile period 3 days prior to ovulation.
Example 2. D~t~ - . . .; .-~l ion of the Ending of Fertile Period This example describes the method of the invention for predicting the end of the fertile period. This example uses the same menstrual cycle as the one used in Example l 20 and measures urinary pregnanediol 3-glucuronide (PDG) assay by the World Health O,ganization radioi--..-.~..o~cs~y method described above.
Example 2(a) This example uses the linear fitting function with the following p~U~ull~l..; 1 25 times minim~l average value, 5 times standard deviation for threshold value, 12%
relative slope, and a score of 3 for declaration of end of fertile period. Table 5 shows the observed concc;"~ion values and other parameters as analyzed for days 11 through 16 accordi~g to the method of the invention.
A four day baseline data is collected for initial relative slope and threshold value wo 95/16920 ~ ~ 7 6 2 ~ 8 PCr/US94/1445S
calculation similar to the procedures described above for Example l. In Example 2(a), day 11 to day 14 provides the four day baseline data of PDG. On day 15, PDG
concentration observed, 11.54 uM, is higher than 1 times minim~l average value and higher than threshold value of the day. Thus, the score is in~ lented by 1. The score is further h~ulcl~el~led by 1 due to a relative slope higher (29.99) than threshold (12) in the base window. However, the ~cunmll~t-ocl score of 2 remains less than that required number for action and, therefore, this day is passed without declaring the end of fertile period. On day 16, observed value is greater than threshold value and score is thus incremented by 1. The resulting i~l~;lc;lllen~al score (3) is the number preset as required for action and day 16 is declared as the end of the fertile period for this particular cycle.
This is three days after ovulation as d~ ...;..P-d by ultrasound (Example 1). Noadditional hormone assays are needed until the be~,h,..i..g of next cycle. At the first day of menses of the next cycle, the procedure for predicting the beginning of fertile period is carried out as ~l~s~ ~ibe~l in Example 1.
Example 2(b) This example uses the third fitting function of ~i ;LII~ ir averaging function.
The pal~lletel~ used are: 5 days after declaration of beginning of fertile period to begin fitting function for ending of fertile pe;iod, a value of 2 times the base value as the threshold value for ending of fertile period, and a score of l to declare action. In this case the multiplier for standard deviation is zero. The baseline data is obtained from day 11 to day 14. The ~ ",~lir average method c~lrlll~te~l the b~cP-line value to 6.5 llM.
The threshold value is then set to 13.0 ~lM (2 times base value; as set in the p~lletel~).
As shown in the Table 5, day 15 showed 11.54 ~M which is lower than the threshold value of 13Ø Therefore day 15 is passed without action. On day 16, the observed PDG value is 13.28 ,uM, i.e., higher than threshold value. The score is in-;lel..~ d to l and the ending of the fertile period is declared because a score of 1 is pre-set for decl~aLion of action. Again, the ending of the fertile period is three days post-ovulation as l.le~ul~d by ultrasound (Example 1).
wo 95/16920 - ~ PCr/uS94/1445 Example 3. Finding the best set of pal~llllGtGl~i for a large data base By collecting and analyzing a large d~t~h~ce co~ lg ~ e.u-ls data points, one can C~/tillli~,G the values ~cign~d to the pre-set pal~llGlGI~, e.g., minim~l average 5 value, relative slope, threshold slope, multiplier, and the score. In this way the method of the invention can be cnctomi7~d to adjust for earlier detection and declaration of beginning of fertile period (for cû--l~a~G~ e utility) or to adjust for later (i.e., closer to ovulation) detection and declaration of beginning of fertile period (for concGp~ion enh~ncement utility).
For this purpose, a large data base with known ovulation day is required. A
collllllonly practiced expG.il--e-,~l design can be used for the ol"i--,i;~alion of ~aldlllclGI~.
Briefly, it involves many G~. h l lr~ runs with regularly varying combination ofpalalllG~Gl~i. The trends of the effect of varying p~alllet~ are analyzed according to the desired relationship to the IGÇ~,lGnce ovulation day. This analysis is then used to predic the best set of parameters for the desired relationship.
Tables 6-8 shows the results from a set of data base collected from 170 cycles.
The time of ovulation of each menstrual cycle is t~ rrninPd by ultrasound investigation to within 24 hours CGI lai--ly. Ovulation is declared when the sonographer notes both a reduction in follicle size and a change in follicle apl)e~dllce by echo pattern and follicle border after rupture. The urinary ElG and urinary PDG assays are done following the RIA method of Examples 1 and 2. The pdlalllGtGI~ tested for this run on this data set are as follows:
For beginning of fertile period:
a minim~l average value of 2.25 times background, 4.5% relative slope, a multiplier of 3.5 times sl~d~d deviation, and a score of 4 for declaration of action.
For end of fertile period:
6 days after declaration of be~,i--l i--g of fertile period to begin fitting function, 2 times minim~l background value, 4.5 times background as threshold - Wo 95/16920 2 1 7 ~ 2 ~ 8 PCT/US94/14455 value, a score of 1, and 11 days as maximal allowable fertile period.
Table 6 shows the results of predicting the be~ g of fertile period of 170 cycles. The first colurnn of the table lists the categories of results relating to ovulation day. For example -9 I~ Stll~ a result that beghlnil,g of fertile period is declared 9 S days before ovulation occurred. The second column pleselll~ the number of cycles that are predicted to have the beginning of fertile period as the co,~ onding number of days from ovulation in the first colurnn. The third column presents the number in the second column as a per~el-~ge fraction of the total number of 170 cycles. The fourth column .~..,sc..~ the cnmlll~t~d fraction of the cycles that have been predicted to have begun fertile period before and on the day shown in the first column before ovulation. The last colurnn represents the 1~ fraction of cycles that have not been announced to have entered fertile period. As can be seen in Table 6, a high majority of cycles (72%) are declared to have begun fertile period 3 days before ovulation and only a small fraction (5%) had not been declared to have entered fertile period by 1 day before ovulation.
Similarly, Table 7 shows the results of testing for the ending of fertile period of 170 cycles. The column and rows of data are arranged and defined as shown in Table 6. As can be seen in the cumlll~tPd fraction column, less than 5% of cycles are announced to have ended fertile period on and before the day of ovulation (as det~ ,I-;--~d by ultrasound), while a majority of cycles (55% to 73%) are announced to have ended the fertile period by the time 4 to 5 days after ovulation.
As can be seen from these results, the predictive value of the method of the invention is superior to other methods known in the art.
Example 4. Comparison to known methods.
As an additional example, the results of the present invention are col,lpd-ed to those published by J.P. Royston, Statistics in Medicine, 10:221-240 (1991). The Royston reference ~ullull~i~ed and ~csPssecl the ~l~llll~lce of various predictive tests of fertile period based on published data. The data are incol~ul~ed into Table 8 and exp~n~l to include data generated by the method of the invention. As is known in the 2176~
wo 95/16920 ` ` PCr/US94/1445 art (see, e.g., pp. 231 of Royston) a commonly accepted criteria for a critical fertility period inclu-les the period of three days pre-ovulation and two days post-ovulation. In Table 8 p~culæltl g is defined as the fraction of cycles that provided a fertile period that overlaps the critical period. P~læter T is an e~ e of the dispersion of the 5 distribution of the fertile period around the ideal value of 6. It is c~l~ulrtç~l by using the following formula:
T = ~ ( f a - 6) 2 + sf 2 } 0-5 10 where fa stands for the average value of the fertile period and Sf stands for the standard deviation of the fertile period.
In Table 8, data from dirr~ sources are colll~ d in terms of the average ~bstin~n~e days and the standard deviation of the sarnples in their respective study in practicing each method. The g p&~ll~,t._l c~ lr~ed above using data ~ t~,d 15 following the method of the invention shows lower values than prior art methods for the reason that all the available data in the sample cycles were used in the c~lculrtion. For a practical use of the method, it would be more a~,u~iate to include those cycles that did not give a~prop,iate signals. Thus, when the cycles classified as not giving a~,op,iate signals are also included into the crlcul~tion, the g values would about 0.61 to 0.64 for 20 the chemical test l and ch~o-rnic~l test 2 in Royston's table. If such g values are used, the values co,l~al~ well with the results ge~ led using the method of the invention (0.61).
The smaller T value which results when using the method of the invention in-lic~t~s that the present invention is capable of giving a much better defined fertile period in a more reproducible manner. Thus, one advantage of the present invention over the prior art 25 methods of predicting fertile period is that a much shorter fertile period is ~sign~1 to each cycle.
Example 5. Trading o~ lion for a shorter predicted fertile period.
Examples 3 and 4 show a set of optimal p~u~net~ which provide good - WO 95/16920 ~ ~ 7 6 2 ~ 8 PCT/US94/14455 p~lro~ ce and an acceptable predicted fertile period. This example shows that the palalllc;~ can be dt;l~ll~ined by following the expelilllt;lllal design used above but with the goal of a shortened predicted fertile period. This provides an additional advantage of fewer testing days which reduces the number of tests which must be taken and provides S a more cost effective and easier to use method.
The o~Lillli,~lion uses a 169 cycle data base from the same participants as used in Example 3, however, one of the 170 cycle data base was ~icqu~lified due to consecutive missed samples half way through the cycle. Ovulation of each menstrual cycle is detelmined by ultrasound investigation to within 24 hours cel laillty as before.However, in this case, the first urine sample collected for analysis is done on day 6 of menses. The urinary E 1 G and urinary PDG assays are done following the RIA method described above. The palalllc;l~l~ are set as follows:
For beginning of fertile period:
Analysis begins on day 6 of menses, no minim~l average value required above background, 4.5% relative slope, a multiplier of 3.5 times standard deviation, and a score of 3.
For end of fertile period:
No minim~l average value required above background, 5% relative slope, a multiplier of 4 times standard deviation, a score of 4, and 10 days as maximal allowed fertile period.
Table 9 shows the results of predicting the beginning of fertile period of these169 cycles. As can be seen in Table 9, a majority of cycles (59%) are declared to have begun fertile period 3 days before actual ovulation (as verified by ultrasound) and only a small fraction of cycles (3.6%) had not been declared to have entered fertile period by the day of ovulation.
Table 10 shows the results of testing for the ending of fertile period of the 169 cycles. As can be seen in the cum~ t~od fraction column, less than 4% of cycles are predicted by the method to have ended fertile period on or before the day of actual ovulation (as ~Ir.l~ "~ Pd by ultrasound), while a majority of cycles (48% to 67%) are Wo 95/16920 PCT/US94/1445~
217 62~8 26 predicted to have ended the fertile period by 4 to 5 days after actual ovulation.
A comparison of the results from Example 3 (the method of the invention commencing on day 2 of menses) and Example 5 (day 6) is shown in Table 11. As can be seen, the pararneters accign~d in Example 5 provide certain advantages over those 5 used in Example 3. The predicted length of fertile phase is reduced from 8.4 to 8.1 days with a conco,~ reduction in standard deviation of fertile phase from 2 to 1.5.
However, this reduction in predicted fertile period results in a reduced index (g) of performance.
The effect of this reduction in g value on the predictive value of the method may 10 be less than the nllm~-rir~l value would indicate since it appears that the reduction is predominately due to a difference of three vs two days pre-ovulation in predicting the start of fertile period. Furthermore, the lower g value may also be offset in part on the presumed use of an early morning urine sample. Therefore, the slightly lower g values which result from col.llll~ncillg the method of the invention on day 6 rather than day 2 15 are well co~ e~ ed by the hlcleased benefit of a shorter predicted fertile period and fewer testing days.
Example 6. Detection of the beginning of fertile period In this example, the begillllillg of fertile period is predicted using a linear fit 20 function and decision rules having the following p~al~ 3% relative slope is set as threshold after day 5,expected value is set by adding 3 times standard deviation to the predicted value, and 3 trigger points are required to declare action. As would be understood, this set of p~lle~ is one of the many sets which may be used to atssess the pelr(,llllà,lce of the method of invention on a large data base. The particular 25 p~alllelel~ used here show the application of the method of the invention and the resulting hlfc,lllldlion which is provided by such data analysis. The l lhl~ulll set of p~al"etel~ for a large data base can be found by pe,rolllli"g a ~lesigned exlJ~lilllelll of system~tiC analysis using known ovulation day for reference v~fir,caLion. Further details of the o~ .d~ion of pal~llel~ is found in Example 8 described below.
zl76248 - wo 95l16920 PCr/US94/14455 In this example the estrogen hormone estrone 3-glucuronide (ElG) is measured in urine. The procedure utilizes a solid phase immllno~c~y on nitrocellulose. In this example in contrast to Example 1, the first ElG assay is done on day 5 instead of day 1 from the start of menses. The advantage of this most p,Grclled method of the invention is that it reduces the number of assays per cycle required to be performed by the user as well as reducing the possibility of urine samples being co~ ted by ~,ellsllual flow.
Again, the sample is plcrcl~bly collected from the first early morning passage of urine.
Samples and assays are obtained and completed for the first four days (i.e., day 5 to day 8 of the menstrual cycle) with assay results from each day read and stored in the meter.
The meter are programmed with the mathematical function of this invention. Thesefirst four days assay results serve as data the baseline data as explained above. As explained above, in the preferred method the meter is programed with function of the invention and thus the actual results are not viewed by the user. Accordingly, the internal operation of the c~l~ ul~tion process inside the meter is ~lesçnhed here in order to more fully explain the method of the invention. The observed data of a first four days results are shown in Table 12.
The observations of Days 5-8 are f tted using the linear function and the other related values described above are derived. Under the pal~llcL,~ set out above, the first four days serve as the baseline days, and, therefore, no trigger count adjustment is made. This results in the minim~l average value being the same as the average value observed because this four day period is the first window observed. Furthermore, there is no predicted value c~lrul~tion and the expected value is not applicable. On Day 9 a daily hormone llle~ulcllltllt is made, the results of which are shown in Table 13.
The working window is now shifted to day 5 to day 9 with abase window of day 5 to day 8. The observed day 9 value (30) is then analyzed ~ccording to the method of the invention. In this case the slope is positive (14) but the observed hollllone assay value (30) is not higher than the minim~l expected value (67.9) and thus there is no trigger point for this feature. However, since the relative slope observed (14) is higher than threshold slope (3) and the day is later than day 8, the trigger point is inclGlllcntcd Wo 95/16920 ~ 1 7 6 2 ~ 8 PCT/US94/14455 by 1. However, the trigger point reached ( 1) is less than set number of 3 and, therefore, day 9 is not the beginning of the fertile period. The process of day 9 is repeated on day 10. The window then shifts to Day 6-10 with a base window of day 6 to day 9. The data observed for day 10 and the results of the method of the invention 5 are shown in the Table 14.
In Table 14 it can be seen that the day 10 observed value (23) is not higher than the expected minim~l average value (66.9), and this is no trigger points are il~;lclllcll~cd by this criterion. Again, since relative slope (17.6) is higher than set threshold (3), the trigger point is hlclclllcllled by 1. The resulting trigger point reached (2) is not higher 10 than trigger number required for action and therefore Day 10 is not declared the beginning of the fertile period. This process is repeated until an action point is reached and the beginning of fertile period is declared. Table 15 shows the proces.cing of data on day 13 and the data from day from 9 to day 13 are displayed.
On day 13, the trigger point reaches the required number for action, namely 3.
15 Thus the beginning of the fertile period for this cycle is declared on day 13. The ovulation day for this particular cycle had been in-lç.pçnr1~ tly established as day 17 by the daily ultrasound observation, i.e., day 17 showed cll~a~;lclistic reduction in follicle di~mr-tçr as well as other features in-lir~ting ovulation such as echo observed in the follicle sac and blurring of follicle borders, etc. Therefore, in this example the method 20 of the invention declares the begilln;llg of fertile period 4 days prior to ovulation.
Example 7. Detç~ ",;n,.lion of the Ending of Fertile Period This example describes the method of the invention for predicting the end of the fertile period and uses the same l"~,nsl,ual cycle as the one used in Example 6. Urinary 25 estrone 3 glucuronide (ElG) is measured by the solid phase immunoassay as desc,ibcd above in Example 6. After the begi""ing of fertile period is decl~ in FY~mple 6 day 13, the daily assay of urinary ElG continues until the pre-set day of day 14. On that day c~lr,ul~tions co,..",~nre using the same running window scheme of five days as that used for pleliclillg the begilmillg of fertile period in Example 6, except the relative - WO95/16920 ~1 762~8 PCrlUS94/1445 slope is c~lcul~t~d from the day of observation itself. In this example the following decision rules are used: default fertile period is 9 days long, reduce fertile period by one day if maximum relative slope is greater than set threshold of 25%, extend fertile period by one day if the beginning of fertile period is less than 10 days while maximum relative S slope is negative, reduce fertile period by 2 days if m~imllm relative slope reached was between day 11 and day 12, reduce fertile period by 1 day if the maximum relative slope is on day 13. Table 16 shows the observed concentration values and other parameters as analyzed for day 9 through 14 according to the method of the invention. As shown in Table 16, the relative slope (42.4%) observed on day 13, is greater than the relative slope of day 14 (13.0). According to the decision rules, a one day reduction infertile period is given for a relative slope greater than threshold (25%) and a fertile period is reduced by one day for reaching maximum on day 13. Therefore, the fertile period for this cycle is 7 days and the end of the fertile period for this cycle is calculated to be day 20. Ovulation day for this menstrual cycle had been detc.lllilled by the ultrasound 15 reference method on day 17 as mentioned above in Example 6. Thus, this declaration of end of fertile period provides 3 additional subsequent days to ovulation for contraception purposes.
Example 8. Determination of Optional P~alllclcl~
By collecting and analyzing a large d~t~h~ce C~ .;Ui.. g llul~crous data points, one can o~ e the values ~c~ign.od to the pre-set pal~llclcl~, e.g., minim~l average value, relative slope, threshold slope, multiplier, and the trigger number. In this way the method of the invention can be cu~ Jllfi~cd to adjust for earlier detection and declaration of beginning of fertile period (for col,ll~cc~ e utility) or to adjust for later 25 (i.e., closer to ovulation) detection and declaration of begi"nillg of fertile period (for conception cl~h~ 'c~e~l utility).
For this purpose, a large data base with known ovulation day is required. A
cornmonly practiced eA~e. ;lll~ design can be used for the u~hl~izalion of p~halllGlcl~..
Briefly, it involves many eA~- - ;",~ runs with regularly varying combination of Wo 95/16920 2 1 7 6 ~ 4 i~ PCr/US9411445~
parameters. The trends of the effect of varying pa,~ne~cl~ are analyzed according to the desired relationship to the reference ovulation day. This analysis is then used to predict the best set of parameters for the desired relationship.
Tables 17-19 shows the results from a set of data base collected from 120 cycles. The time of ovulation of each menstrual cycle is dete- ,II;II~d by ultrasound investigation to within 24 hours certainty. Ovulation is declared when the sonographer notes both a reduction in follicle size and a change in follicle a~pe~dllce by echo pattern and follicle border after rupture. The urinary ElG assay çc.centi~lly follows the solid phase immunoassay as tlPs~-rihecl in Examples 6 and 7. The p~ulle~.s tested for this Example on this data set are as the same as described in Example 6 and 7.
Table 17 shows the results of predicting the beginning of fertile period for 120cycles. The first column of the table lists the results obtained by the method of the invention as cOlll~alcd to actual ovulation day. For example 4 ,c~lcsenL~ a result that the present method declares the beginning of fertile period 4 days before a confined ovulation had occurred. The second column plcs~.lL~ the number of cycles that are predicted to have the beginning of fertile period as the coll~s~ondillg number of days from ovulation in the first column. The third column plcScllL~ the number in the second column as a pel-;clll~ge fraction of the total number of 120 cycles. The fourth column Ic~lcScllL~ the cum~ tecl fraction of the cycles that have been predicted to have begun fertile period before and on the day shown in the first column beforc ovulation. The last column lc~lcscll~s the lClll~ g fraction of cycles that have not been announced to have entered fertile period. As can be seen in Table 17, a high majority of cycles (66%) are declared to have begun fertile period 3 days before ovulation and only a small fraction (8%) had not been declared to have entered fertile period by I day before ovulation.
Similarly, Table 18 shows the results of testing for the ending of fertile period of 170 cycles. The data is arranged and defined as in Table 17. As can be seen in the cum~ ted fraction column, less than 8% of cycles are announced to have ended fertile period on and before the day of ovulation (as detel"l",ed by ultrasound), while a majority of cycles (43% to 59%) are announced to have ended the fertile period by the 2~76248 --- WO 9S/16920 Pcr/uss4/l4455 time 4 to 5 days after ovulation.
As can be seen from these results, the predictive value of the method of the invention is superior to other methods known in the art.
5 Example 9. Co~ alisol1 to known methods.
As a further collfi,llla~ion of the superior predictive use of the present invention, the results of the present invention are compared to those published by J.P. Royston, Statistics in Medicine, 10:221-240 (1991). The Royston reference ~.ulll~l~izes and assesses the performance of various predictive tests of fertile period based on published lO data. The data are incorporated into Table 19 and expanded to include data generated by the method of the invention. As is known in the art (see, e.g., pp. 231 of Royston) a commonly accepted criteria for a critical fertility period includes the period of three days pre-ovulation and two days post-ovulation. In Table 19 parameter g is defined as the fraction of cycles that provided a fertile period that overlaps the critical period.
15 Parameter T is an çstim~ts of the dispersion of the distribution of the fertile period around the ideal value of 6. It is calc~ tP-d by using the following formula:
T={(f - 6)2+ sf 2}0.5 20 where fa stands for the average value of the fertile period and Sf stands for the standard deviation of the fertile period.
In Table 19, data from dirr~.Gnt sources are colllp~;d in terms of the average abstinence days and the standard deviation of the Salllpl-~ in theimcs~ ;Li~e study in practicing each method. The g pdl.:ullt;Lcl c~lrlll~tecl above using data generated 25 following the method of the invention shows lower values than prior art methods for the reason that all the available data in the sample cycles were used in the c~lr,~l~ltion. For a practical use of the method, it would be more a~ ,iate to include those cycles that did not give ap~"o~liate signals. Thus, when the cycles cl~cci~lec~ as not giving a~pl~,~liate signals are also included into the s~lrlll~tion7 the g values would approximately 0.65 to Wo 95/16920 ;~ 17 6 2 4 ~` PCT/US94/14455 0.68 for the either chemical test l or chemical test 2. If such g values are used, the values co~ well with the results generated using the method of the present invention (0.68). The smaller T value which results when using the method of the inventioninrlir~tes that the present invention is capable of giving a much better defined fertile 5 period in a more reproducible manner. Thus, one advantage of the present invention over the prior art methods of predicting fertile period is that a much shorter fertile period is assigned to each cycle.
As can be seen from these results, the predictive value of the method of the invention is superior to other methods known in the art.
Examples 8 and 9 show a set of optimal parameters which provide good performance and an acceptable predicted fertile period. As is understood by those skilled in the art, the parameters can be ~letermin~od by following the ex~.i",~ l design used above but with the goal of a shortened predicted fertile period. By applying the principle of.design of e~ , another set of ~alal~le~ could be devised to 15 provide an additional advantage of fewer testing days which reduces the number of tests which must be taken and provides a more cost effective and easier to use method. On the other hand, another set of parameters, similar to Examples 8, could be devised to provide higher protection with the understood expense in running more assays for a longer period.
The alternative emborlimrnt~ described are intrn~ d as examples rather than as limits. Thus, the description of the invention is not intrn-lPd to limit the invention to the particular embo-l;",~ ; disclosed, but it is inten-led to enco",~ass all equivalents and subject matter within the spirit and scope of the invention as observed above and as set forth in the following claims.
WO 95116920 2 1 7 6 2 ~ 8 PCr/US94/14455 TABLE 1- Day 5 Cycle Assay Relative Standard Minimal Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 2 62.61
3 58.76
4 75.33 108.7 20.28 76.35 N/A N/A 0 Note: N/A stands for not applicable.
TABLE 2- Day 6 Cycle Assay Relative Standard Minimal Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 2 62.61 3 58.76 4 75.33 108.7 20.28 10.75 76.35 N/A N/A 0 6 89.35 20.28 10.75 76.35 115.06 168.81 0 TABLE 3- Day 7 Cycle Assay Relative Standard Mini~ Predicted FYpect~d Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 3 58.76 4 75.33 108.7 20.28 10.75 76.35 N/A N/A 0 6 89.35 20.28 10.75 76.35 115.06 999999 7 120.37 15.07 13.71 76.35 114.32 168.81 2 WO 95/16920 ~ 2 ~ 8 PCTIUS94/1445S
TABLF 4- Day 10 Cycle Assay Relative Standard MiniIT~ Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 6 89.35 20.28 10.75 76.35 115.06 999999 7 120.37 15.07 13.71 76.35 114.32 168.81 2 8 99.03 11.76 13.33 76.35 127.38 182.89 3 9 162.64 0.19 13.27 76.35 104.87 194.01 3 182.20 10.83 19.36 76.35 167.48 171.22 4 TABLE 5- Day 16 Cycle Assay Relative Standard Mini~ Predicted Expected Trigger Day PDG Slope Deviation Average Value Value Points (uM) (%) Value 11 2.77 12 5.25 13 5.39 14 12.6 45.57 0 11.54 45.57 1.83 6.50 13.91 8.23 2 16 13.28 29.99 2.01 6.50 15.21 23.05 3 2~7624~
Daysfrom No. Cycle Fraction (%) of Cum~ t~d R~rn~ining ovulation predicted to begin total 170 cycles Fraction (%) Fractions (%) DayFertile period S-9 15 8.8 8.8 91.2 -8 6 3.5 12.3 87.7 -7 9 5.3 17.6 82.4 -6 11 6.5 24.1 75.9
TABLE 2- Day 6 Cycle Assay Relative Standard Minimal Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 2 62.61 3 58.76 4 75.33 108.7 20.28 10.75 76.35 N/A N/A 0 6 89.35 20.28 10.75 76.35 115.06 168.81 0 TABLE 3- Day 7 Cycle Assay Relative Standard Mini~ Predicted FYpect~d Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 3 58.76 4 75.33 108.7 20.28 10.75 76.35 N/A N/A 0 6 89.35 20.28 10.75 76.35 115.06 999999 7 120.37 15.07 13.71 76.35 114.32 168.81 2 WO 95/16920 ~ 2 ~ 8 PCTIUS94/1445S
TABLF 4- Day 10 Cycle Assay Relative Standard MiniIT~ Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 6 89.35 20.28 10.75 76.35 115.06 999999 7 120.37 15.07 13.71 76.35 114.32 168.81 2 8 99.03 11.76 13.33 76.35 127.38 182.89 3 9 162.64 0.19 13.27 76.35 104.87 194.01 3 182.20 10.83 19.36 76.35 167.48 171.22 4 TABLE 5- Day 16 Cycle Assay Relative Standard Mini~ Predicted Expected Trigger Day PDG Slope Deviation Average Value Value Points (uM) (%) Value 11 2.77 12 5.25 13 5.39 14 12.6 45.57 0 11.54 45.57 1.83 6.50 13.91 8.23 2 16 13.28 29.99 2.01 6.50 15.21 23.05 3 2~7624~
Daysfrom No. Cycle Fraction (%) of Cum~ t~d R~rn~ining ovulation predicted to begin total 170 cycles Fraction (%) Fractions (%) DayFertile period S-9 15 8.8 8.8 91.2 -8 6 3.5 12.3 87.7 -7 9 5.3 17.6 82.4 -6 11 6.5 24.1 75.9
-5 26 15.3 39.4 60.6 -4 29 17.1 56.5 43.5 -3 27 15.9 72.4 27.6 -2 20 11.8 84.2 15.8 -1 18 10.6 94.8 5.2 7 4.1 98.9 1.1 0.6 99.5 0.5 > 2 1 0.6 100.0 0.0 WO 95/16920 2 ~ 6 2 ~ PCr/US94114455 Daysfrom No. Cycle Fraction (%) of~um~ t~o-dRemaining ovulationpredicted to end total 170 cycles Fraction (%) Fractions (%) Day Fertile period <-2 1 0.6 0.6 99.4 - 1 2 1.2 1.8 98.2 0 5 2.9 4.7 95.3 11 6.5 11.2 88.8 2 22 12.9 24.1 75.9 3 27 15.9 40.0 60.0 4 25 14.7 54.7 45.3 31 18.2 72.9 27.1
6 21 12.4 85.3 14.7
7 9 5.3 90.6 9.4
8 7 4.1 94.7 5.3 29 9 5.3 100.0 0.0 Wo 95/16920 ~ 1 7 ~ ~ ~ g ` PCr/US94/14455 Length of Indices of fertile phase pelrol~ ce Mean SD
Method (days) (days~ g T
Sympto-thermal 13.4 2.9 0.98 7.9 Cervical Mucus (billings)11.9 2.9 0.91 6.6 Thermal (C~lrul~tion and BBT) 11.8 3.3 0.90 6.7 Ch~-mic~l test 1 9.3 2.2 0.83 4.0 (~h.-.mic~l test2 10.7 2.3 0.84 5.2 This invention 8.4 2.0 0.48a 3.2 This invention 8.4 2.0 0.61b 3.2 Note: a: c~lrul~tr.d with all cycles using -3 and +3 from ovulation as criterion.
b: c~lrul~t~d with all cycles using -3 and +2 from ovulation as criterion.
WO 95/169202 l 7 6 2 4 8 PCr/US94/1445S
DaysfromNo. Cycle Fraction (%) of Cum~ tPd R~om~inin~
ovulationpredicted to begin total 169 cyclesFraction (%)Fractions (%) DayFertile period S -9 11 6.5 6.5 93.5 -8 5 3.0 9.5 90.5 -7 4 2.4 11.9 88.1 -6 8 4.7 16.6 83.4 -5 17 10.1 26.7 73.3 -4 24 14.2 40.9 59.1 -3 31 18.3 59.2 40.8 -2 31 18.3 77.5 22.5 -1 25 14.8 92.3 7.7 0 7 4.1 96.4 3.6 3.0 97.4 0.6 2 2 1 0.6 100.0 0.0 --WO95/16920 ~ 76~248 PCT/US94/14455 Daysfrom No. Cycle Fraction (%) of Cum~ !n~inin~
ovulation predicted to end total 169 cycles Fraction (%) Fractions (%) Day Fertile period ~-2 2 1.2 1.2 98.8 -1 4 2.4 3.6 96.4 0 6 3.6 7.2 92.8 7 4.1 11.3 88.7 2 13 7.7 19.0 81.0 3 18 10.7 29.7 70.3 4 31 18.3 48.0 52.0 32 18.9 66.9 33.1 6 21 12.4 79.3 20.7 7 13 7.7 87.0 13.0 8 8 4.7 91.7 8.3 >9 14 8.3 100.0 0.0 WO 95/16920 2 1 7 ~ 2 4 8 PCrlUS94/1445S
~40 Predicted length of Indices of fertile period pe,Çol",al~ce Mean SD
Method (days) (days) g T
Example 3 8.4 2.0 0.48a 3.2 Example 3 8.4 2.0 0.61b 3.2 Example 5 8.1 1.5 0.40a 2.5 Example 5 8.1 1.5 0.48b 2.5 Note: a: c~l~ul~t~1 with all cycles using -3 and +3 from ovulation as criterion.b: calculated with all cycles using -3 and +2 from ovulation as criterion.
-Wo 95/16920 2 1 ~ 6 2 ~ 8 PCrlUS94/14455 TABLE 12- Day 5 to Day 8 Basel: ne Region Data Cycle Assay Relative Standard Minirnal Predicted ExI~ecte~l Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 8 40 4.2 9.13 30 N/A N/A 0 Note: N/A stands for not applicable.
TABLE 13- Day 9 Cycle Assay Relative Standard Mini~ Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 8 40 14 9.13 30 N/A N/A 0
Method (days) (days~ g T
Sympto-thermal 13.4 2.9 0.98 7.9 Cervical Mucus (billings)11.9 2.9 0.91 6.6 Thermal (C~lrul~tion and BBT) 11.8 3.3 0.90 6.7 Ch~-mic~l test 1 9.3 2.2 0.83 4.0 (~h.-.mic~l test2 10.7 2.3 0.84 5.2 This invention 8.4 2.0 0.48a 3.2 This invention 8.4 2.0 0.61b 3.2 Note: a: c~lrul~tr.d with all cycles using -3 and +3 from ovulation as criterion.
b: c~lrul~t~d with all cycles using -3 and +2 from ovulation as criterion.
WO 95/169202 l 7 6 2 4 8 PCr/US94/1445S
DaysfromNo. Cycle Fraction (%) of Cum~ tPd R~om~inin~
ovulationpredicted to begin total 169 cyclesFraction (%)Fractions (%) DayFertile period S -9 11 6.5 6.5 93.5 -8 5 3.0 9.5 90.5 -7 4 2.4 11.9 88.1 -6 8 4.7 16.6 83.4 -5 17 10.1 26.7 73.3 -4 24 14.2 40.9 59.1 -3 31 18.3 59.2 40.8 -2 31 18.3 77.5 22.5 -1 25 14.8 92.3 7.7 0 7 4.1 96.4 3.6 3.0 97.4 0.6 2 2 1 0.6 100.0 0.0 --WO95/16920 ~ 76~248 PCT/US94/14455 Daysfrom No. Cycle Fraction (%) of Cum~ !n~inin~
ovulation predicted to end total 169 cycles Fraction (%) Fractions (%) Day Fertile period ~-2 2 1.2 1.2 98.8 -1 4 2.4 3.6 96.4 0 6 3.6 7.2 92.8 7 4.1 11.3 88.7 2 13 7.7 19.0 81.0 3 18 10.7 29.7 70.3 4 31 18.3 48.0 52.0 32 18.9 66.9 33.1 6 21 12.4 79.3 20.7 7 13 7.7 87.0 13.0 8 8 4.7 91.7 8.3 >9 14 8.3 100.0 0.0 WO 95/16920 2 1 7 ~ 2 4 8 PCrlUS94/1445S
~40 Predicted length of Indices of fertile period pe,Çol",al~ce Mean SD
Method (days) (days) g T
Example 3 8.4 2.0 0.48a 3.2 Example 3 8.4 2.0 0.61b 3.2 Example 5 8.1 1.5 0.40a 2.5 Example 5 8.1 1.5 0.48b 2.5 Note: a: c~l~ul~t~1 with all cycles using -3 and +3 from ovulation as criterion.b: calculated with all cycles using -3 and +2 from ovulation as criterion.
-Wo 95/16920 2 1 ~ 6 2 ~ 8 PCrlUS94/14455 TABLE 12- Day 5 to Day 8 Basel: ne Region Data Cycle Assay Relative Standard Minirnal Predicted ExI~ecte~l Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 8 40 4.2 9.13 30 N/A N/A 0 Note: N/A stands for not applicable.
TABLE 13- Day 9 Cycle Assay Relative Standard Mini~ Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 8 40 14 9.13 30 N/A N/A 0
9 30 14 9.13 30 40.5 67.9 TABLE 14- Day 10 Cycle Assay Relative Standard Minin~ Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 8 40 14 9.1 30 N/A N/A 0 9 30 14 9.1 30 40.5 67.9 23 17.6 8.1 29.5 42.5 66.9 2 Wo 95116920 ~ 17 6 2 ~ 8 PCT/US94/14455 TABLE 15- Day 13 Cycle Assay Relative Standard Minimal Predicted Expected Trigger Day ElG Slope Deviation Average Value Value Points (nM) (%) Value 9 30 14 9.1 29.5 40.5 67.9 23 17.6 8.1 29.5 42.5 66.9 2 11 37 -12.7 4.8 29.5 21.5 999999 2 12 84 -4.9 7.3 29.5 28.5 999999 2 13 90 40.5 15.7 29.5 87.5 134.6 3 TABLE 16- Day 13 and Day 14 Cycle Assay Relative Standard Minimal Predicted FY~t~d Trigger Day PDG Slope Deviation Average Value Value Points (uM) (%) Value 13 90 42.4 15.7 29.5 87.5 134.6 3 14 65 13.0 2.01 6.50 15.21 23.05 3 --WO 95/16920 ~ 1 7 6 2 4 8 PCT/US94/14455 Days from No. Cycle Fraction (%) ofC'um~ t~d Rem~ining co~ -,Fd predicted to begin total 120 cycles Fraction (%) Fractions (%) ovulation (1*) Fertileperiod 2-9 7 5.8 5.8 94.2 -8 5 4.2 10.0 90.0 -7 2 1.7 11.7 88.3 -6 7 5.8 17.5 82.5 -5 21 17.5 35.0 65.0 -4 14 11.7 46.7 53.3 -3 23 19.2 65.8 34.2 -2 16 13.3 79.2 20.8 -1 15 12.5 91.7 8.3 0 7 5.8 97.5 2.5 2 1.7 99.2 0.8 2 2 1 0.8 100.0 0.0 1 * = Confirmed by ultrasound, see text.
Wo 95/16920 i PCT/US94/1445~-i~ l 7 ~
Daysfrom No. CycleFraction (%)of Cum~ ted Remaining conrllllled predictedtoend total 120cycles Fraction(%) Fractions (%) ovulation (1 *) Fertile period <-2 3 2.5 2.5 97.5 -1 3 2.5 5.0 95.0 4 3.3 8.3 91.7 3 2.5 10.8 89.2 2 4 3.3 14.2 85.8 3 11 9.2 23.3 76.7 4 23 19.2 42.5 57.5 16.7 59.2 40.8 6 18 15.0 74.2 25.8 7 15 12.5 86.7 13.3 8 10 8.3 95.0 5.0 2 9 6 5.0 100.0 0.0 I * = Confirmed by ultrasound, see text.
WO 95tl6920 ~ 1 7 6 2 4 ~ PCr/US94/14455 Length of Indices of fertile phase ~tlÇollll~ce Mean SD
Method (days) (days) g T
Sympto-thermal 13.4 2.9 0.98 7.9 Cervical Mucus (billings) 11.9 2.9 0.91 6.6 Thermal (~ tion and BBT) 11.8 3.3 0.90 6.7 Chemical test 1 9.3 2.2 0.83 4.0 Ch~nni~l test 2 10.7 2.3 0.84 5.2 This invention 8.4 2.0 0.65a 2.7 This invention 8.4 2.0 0.68b 2.7 Note: a: c~lrul~t~d with all cycles using -3 and +3 from ovulation as criterion.b: C~lCUl~t.q.~l with all cycles using -3 and +2 from ovulation as criterion.
Wo 95/16920 i PCT/US94/1445~-i~ l 7 ~
Daysfrom No. CycleFraction (%)of Cum~ ted Remaining conrllllled predictedtoend total 120cycles Fraction(%) Fractions (%) ovulation (1 *) Fertile period <-2 3 2.5 2.5 97.5 -1 3 2.5 5.0 95.0 4 3.3 8.3 91.7 3 2.5 10.8 89.2 2 4 3.3 14.2 85.8 3 11 9.2 23.3 76.7 4 23 19.2 42.5 57.5 16.7 59.2 40.8 6 18 15.0 74.2 25.8 7 15 12.5 86.7 13.3 8 10 8.3 95.0 5.0 2 9 6 5.0 100.0 0.0 I * = Confirmed by ultrasound, see text.
WO 95tl6920 ~ 1 7 6 2 4 ~ PCr/US94/14455 Length of Indices of fertile phase ~tlÇollll~ce Mean SD
Method (days) (days) g T
Sympto-thermal 13.4 2.9 0.98 7.9 Cervical Mucus (billings) 11.9 2.9 0.91 6.6 Thermal (~ tion and BBT) 11.8 3.3 0.90 6.7 Chemical test 1 9.3 2.2 0.83 4.0 Ch~nni~l test 2 10.7 2.3 0.84 5.2 This invention 8.4 2.0 0.65a 2.7 This invention 8.4 2.0 0.68b 2.7 Note: a: c~lrul~t~d with all cycles using -3 and +3 from ovulation as criterion.b: C~lCUl~t.q.~l with all cycles using -3 and +2 from ovulation as criterion.
Claims (13)
1. A method for determining the periodic fertile period in a female, the method comprising the steps of:
(a) collecting a daily sample of biological fluid from the female commencing start of a fixed number of days from the menses and measuring a hormone level in the daily sample for a number of consecutive days;
(b) inputting the measured hormone level and corresponding day from the first day of menses into a data storage means;
(c) analyzing the hormone level using a computational means, the computational means comprising the application of a fitting function, extrapolating a threshold value of hormone level from the fitted function, comparing the measured hormone level and the threshold value using a scoring means, the scoring means defining preset relationship between the measured hormone level and threshold value;
(d) defining a first day of the fertile period and a final day of the fertile period as the day when application of the scoring means results in an accumulated score greater than a preset value; and (e) displaying a predicted first day of the fertile period and a predicted final day of the fertile period on an output means;
whereby the female is considered capable of conception from the time of reaching the first day of fertile period until the final day of the fertile period.
(a) collecting a daily sample of biological fluid from the female commencing start of a fixed number of days from the menses and measuring a hormone level in the daily sample for a number of consecutive days;
(b) inputting the measured hormone level and corresponding day from the first day of menses into a data storage means;
(c) analyzing the hormone level using a computational means, the computational means comprising the application of a fitting function, extrapolating a threshold value of hormone level from the fitted function, comparing the measured hormone level and the threshold value using a scoring means, the scoring means defining preset relationship between the measured hormone level and threshold value;
(d) defining a first day of the fertile period and a final day of the fertile period as the day when application of the scoring means results in an accumulated score greater than a preset value; and (e) displaying a predicted first day of the fertile period and a predicted final day of the fertile period on an output means;
whereby the female is considered capable of conception from the time of reaching the first day of fertile period until the final day of the fertile period.
2. A method of claim 1 further comprising the measurement of a first hormone for a first period commencing a fixed number of days from the start of menses and a second hormone for a second period commencing the end of the first period.
3. The method of claim 2 wherein the first hormone is estrogen or an estrogenmetabolite and the second hormone is progesterone or progesterone metabolite.
4. The method of claim 3 wherein collecting the first period commences the second day of menses.
5. The method of claim 3 wherein collecting the first period commences the sixth day of menses.
6. A method for determining the periodic fertile period in a female, the method comprising the steps of:
(a) measuring a hormone level in a daily sample of a biological fluid of the female for a testing period commencing on a fixed number of days from the start of menses;
b) storing the measured hormone level and corresponding day from start of menses into a data storage means;
(c) analyzing the hormone level using a computational means, the computational means comprising a microprocessor for applying a fitting function to normalized data;
(d) processing analyzed data in a regime of three stages, including a baseline stage, a predicting stage and a confirming stage;
(e) extrapolating an expected hormone level from the fitted function;
(f) the predicting stage comparing measured hormone level to extrapolated hormone level using a scoring means, the scoring means defining a preset relationship between measured and extrapolated hormone values;
(g) designating the first day of the fertile period as the day when accumulated scores in the predicting stage reach or exceed a predetermined value;
(h) the confirming stage storing the measured hormone level from the first day of the fertile period until the end of the testing period;
(i) analyzing the stored hormone data at the end of the testing period with a second scoring means comprising a preset decision rule;
(j) designating the last day of the fertile period based on the outcome of the second scoring means; and (k) displaying the first and last days of the fertile period on an output means;
whereby the female is considered capable of conception from the time of reaching the first day of the fertile period until the last day of the fertile period.
(a) measuring a hormone level in a daily sample of a biological fluid of the female for a testing period commencing on a fixed number of days from the start of menses;
b) storing the measured hormone level and corresponding day from start of menses into a data storage means;
(c) analyzing the hormone level using a computational means, the computational means comprising a microprocessor for applying a fitting function to normalized data;
(d) processing analyzed data in a regime of three stages, including a baseline stage, a predicting stage and a confirming stage;
(e) extrapolating an expected hormone level from the fitted function;
(f) the predicting stage comparing measured hormone level to extrapolated hormone level using a scoring means, the scoring means defining a preset relationship between measured and extrapolated hormone values;
(g) designating the first day of the fertile period as the day when accumulated scores in the predicting stage reach or exceed a predetermined value;
(h) the confirming stage storing the measured hormone level from the first day of the fertile period until the end of the testing period;
(i) analyzing the stored hormone data at the end of the testing period with a second scoring means comprising a preset decision rule;
(j) designating the last day of the fertile period based on the outcome of the second scoring means; and (k) displaying the first and last days of the fertile period on an output means;
whereby the female is considered capable of conception from the time of reaching the first day of the fertile period until the last day of the fertile period.
7. A method for determining the beginning of the periodic fertile period in a female the method comprising the steps of:
(a) collecting a daily sample of biological fluid from the female commencing on a pre-selected fixed day after start of menses and continuing through a designated first day of the fertile period;
(b) measuring estrogen-related hormone level in the sample;
(c) storing the measured hormone level and corresponding day from start of menses on which the hormone level is measured into a data storage cans;
(d) analyzing the hormone level using a computational means, the computational means composing a microprocessor for applying a fitting function to normalized data; such that the computational means calculates a trend in hormone level variations and determines expected hormone level levels based on a trend in measured hormone level variations;
(e) comparing measured and extrapolated hormone levels using a scoring means;
(f) designating the first day of said fertile period when application of the scoring means results in an accumulated score greater than a preset value; and (g) displaying a result indicating that the female is considered capable of conception after the first day of the fertile period.
(a) collecting a daily sample of biological fluid from the female commencing on a pre-selected fixed day after start of menses and continuing through a designated first day of the fertile period;
(b) measuring estrogen-related hormone level in the sample;
(c) storing the measured hormone level and corresponding day from start of menses on which the hormone level is measured into a data storage cans;
(d) analyzing the hormone level using a computational means, the computational means composing a microprocessor for applying a fitting function to normalized data; such that the computational means calculates a trend in hormone level variations and determines expected hormone level levels based on a trend in measured hormone level variations;
(e) comparing measured and extrapolated hormone levels using a scoring means;
(f) designating the first day of said fertile period when application of the scoring means results in an accumulated score greater than a preset value; and (g) displaying a result indicating that the female is considered capable of conception after the first day of the fertile period.
8. A method for determining the end of the periodic fertile period in a female, the method comprising the steps of:
(a) collecting a daily sample of a biological fluid from the female commencing on a fixed day of the female menstrual period and continuing for a fixed number of days of menstrual period;
(b) measuring an estrone-related hormone level in the sample;
(c) storing the estrone-related hormone level and corresponding day from start of menses on which the hormone level was measured into a means for computting a trend in daily variations in measured hormone levels;
(d) scoring variations in daily measured estrone-related hormone levels using a scoring means; and (f) designating the final day of the fertile period when the scoring means results in an outcome of a preset decision rules; whereby the female is considered capable of conception from the time of reaching the first day of fertile period until the final day of the fertile period.
(a) collecting a daily sample of a biological fluid from the female commencing on a fixed day of the female menstrual period and continuing for a fixed number of days of menstrual period;
(b) measuring an estrone-related hormone level in the sample;
(c) storing the estrone-related hormone level and corresponding day from start of menses on which the hormone level was measured into a means for computting a trend in daily variations in measured hormone levels;
(d) scoring variations in daily measured estrone-related hormone levels using a scoring means; and (f) designating the final day of the fertile period when the scoring means results in an outcome of a preset decision rules; whereby the female is considered capable of conception from the time of reaching the first day of fertile period until the final day of the fertile period.
9. A method according to claim 6 wherein the testing period commences on day 5.
10. A method according to claim 6 wherein the testing period commences on day 2.
11. A method according to claim 6 wherein the baseline is 4 days.
12. A method according to claim 6, wherein the testing period is 10 days.
13. A method according to claim 6 wherein the hormone is estrone 3-glucuronide (E1G).
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US16854893A | 1993-12-16 | 1993-12-16 | |
| US08/168,548 | 1993-12-16 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2176248A1 true CA2176248A1 (en) | 1995-06-22 |
Family
ID=22611955
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002176248A Abandoned CA2176248A1 (en) | 1993-12-16 | 1994-12-16 | Methods and means for determining the female fertile period |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0734531A1 (en) |
| JP (1) | JPH09506713A (en) |
| AU (1) | AU1402295A (en) |
| CA (1) | CA2176248A1 (en) |
| WO (1) | WO1995016920A1 (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9217864D0 (en) | 1992-08-21 | 1992-10-07 | Unilever Plc | Monitoring method |
| IL140394A0 (en) * | 1998-06-22 | 2002-02-10 | Medi Cult As | A method for in vitro maturation of human gametes |
| US7427271B2 (en) * | 2002-07-01 | 2008-09-23 | Vaclav Kirsner | Diagnosis of fertility status by folliculogenesis monitoring in the vagina |
| JP4150323B2 (en) * | 2003-10-29 | 2008-09-17 | 浜松ホトニクス株式会社 | Immunochromatographic test strip reader, cartridge used therefor, and immunochromatographic test strip inspection system |
| US9939385B2 (en) | 2011-09-09 | 2018-04-10 | Church & Dwight Co., Inc. | Systems, methods, and test kits for analyte variation detection |
| RU2019100456A (en) * | 2016-06-13 | 2020-07-14 | Конинклейке Филипс Н.В. | OPTICAL MONITORING OF FACE CHARACTERISTICS DURING A WOMAN'S MENSTRUAL CYCLE |
| US11029321B2 (en) | 2017-02-17 | 2021-06-08 | MFB Fertility, Inc. | Method of evaluating corpus luteum function by recurrently evaluating progesterone non-serum bodily fluids on multiple days |
| US11061026B2 (en) | 2017-02-17 | 2021-07-13 | MFB Fertility, Inc. | System of evaluating corpus luteum function by recurrently evaluating progesterone non-serum bodily fluids on multiple days |
| WO2021034412A1 (en) * | 2019-08-19 | 2021-02-25 | MFB Fertility, Inc. | Systems and methods for menstrual cycle testing |
| US11131665B1 (en) | 2018-08-22 | 2021-09-28 | MFB Fertility, Inc. | Method for evaluating urine of a subject to estimate the fertile window |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2029011B (en) * | 1978-09-01 | 1983-02-02 | Coulson W | Use of a synthetic bifunctional ligand for the immunimetric determination of the concentration ratio of two solutes |
| DE3112767A1 (en) * | 1981-03-31 | 1982-11-11 | Werner Dr.med. 1000 Berlin Steinschulte | Home computer for natural family planning (NFP) and for gynaecological-endocrinological diagnosis |
| IE54109B1 (en) * | 1982-02-10 | 1989-06-21 | Boots Celltech Diagnostics | Assay |
| US5118630A (en) * | 1988-11-04 | 1992-06-02 | Quidel Corporation | Method for determining periodic infertility in females |
| EP0606343A1 (en) * | 1991-10-01 | 1994-07-20 | BIGGEL, Emil Josef | Menstrual cycle meter |
-
1994
- 1994-12-16 CA CA002176248A patent/CA2176248A1/en not_active Abandoned
- 1994-12-16 EP EP95905393A patent/EP0734531A1/en not_active Withdrawn
- 1994-12-16 AU AU14022/95A patent/AU1402295A/en not_active Abandoned
- 1994-12-16 JP JP7516950A patent/JPH09506713A/en active Pending
- 1994-12-16 WO PCT/US1994/014455 patent/WO1995016920A1/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| JPH09506713A (en) | 1997-06-30 |
| AU1402295A (en) | 1995-07-03 |
| EP0734531A1 (en) | 1996-10-02 |
| WO1995016920A1 (en) | 1995-06-22 |
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