WO2010061613A1

WO2010061613A1 - Method, system and kit for supporting pregnancy

Info

Publication number: WO2010061613A1
Application number: PCT/JP2009/006399
Authority: WO
Inventors: 穴沢隆; 藤田毅; 矢澤義昭
Original assignee: 株式会社日立製作所
Priority date: 2008-11-28
Filing date: 2009-11-26
Publication date: 2010-06-03
Also published as: JPWO2010061613A1; JP5308452B2

Abstract

Provided are a method and a system for enabling highly accurate estimation of LH surge peak time and ovulation time at the early stage. A method which comprises: collecting urine samples of a single subject at intervals of 24 hours during a single menstruation cycle; determining luteinizing hormone (LH) in the samples to give LH concentrations (quantitative values) (a diamond plot); at the point (48 hours of the sampling) when the LH concentration exceeds a predetermined LH concentration threshold (60 mlU/ml), fitting a predetermined LH concentration time-change function (Gauss distribution) for the LH concentrations (quantitative values) obtained until that point (three diamond plots indicated by the arrows) using the least squares method; estimating the peak time of the function after the fitting as the LH surge peak time (69 hours of the sampling); and estimating the ovulation time based on the LH surge peak time. The dotted line indicates the correct LH surge peak time (75 hours of the sampling).

Description

Pregnancy support method, system and kit

The present invention relates to an analysis system and method for analyzing biological substances such as nucleic acids, proteins and microorganisms contained in biological samples such as blood, urine and saliva. In particular, the present invention relates to a monitoring method, a monitoring system, and a test kit for analyzing luteinizing hormone contained in urine for the purpose of estimating ovulation time for supporting pregnancy of women.

Declining birthrate is one of the most important national issues. In 2005, Japan's total fertility rate reached 1.26 (1.06 million births), the lowest after the war, and entered a society with a declining population. The main causes are women's social advancement, unmarried women, late marriage and late birth due to economic independence. On the other hand, these are serious issues for women. Not only the social and economic problems of the person, but also the problem of infertility that you cannot get pregnant if you want. According to Non-Patent Document 1, about 10% of couples of the age (reproductive age) that children can make, 1.4 million couples do not have children, and half of them 700,000 couples are worried about infertility Yes (National Institute of Population and Social Security Research Basic Survey on Birth Trends). On the other hand, only 300,000 couples are actually receiving infertility treatment (estimated by the Ministry of Health, Labor and Welfare), and half 150,000 couples have children (Nikkei Medical Estimate). Reasons for not receiving infertility treatment are: “I don't want to have a child until after treatment in the hospital”, “Infertility treatment costs money”, “I do n’t agree with my husband ’s feelings about having a child,” “ I don't know which hospital I should go to. " For working women, it is difficult to achieve a balance between work and treatment, and the situation is more serious, such as being forced to choose between these two options (Non-patent Document 2).

不 Infertility treatment mainly includes (1) timing therapy, (2) hormone therapy, (3) artificial insemination, and (4) in vitro fertilization, which are generally performed while stepping up in this order. For example, perform (1) for the first 3 months, and if you do not become pregnant at this stage, perform (2) while using (1) for the next 3 months. (1) to (3) are also called general fertility treatments, and (4) is also called advanced fertility treatments. According to Non-Patent Document 1 and the homepages of hospitals that conduct infertility treatment, the statistical proportion of successful pregnancy steps is 14% for (1), 33% for (2), and (3) Is 19% and (4) is around 34%.

If the same level can be easily implemented at home without going to the hospital (1), 400,000 couples who are concerned about infertility and who have not received infertility treatment will・ Economical / physical barriers will be lower, making it easier to work on (1) and potentially being saved. If all 400,000 couples work on (1), it is expected that 400,000 couples x 50% x 14% = 30,000 couples will become pregnant. This number is statistically linked to the increase in the number of births. In addition, the remaining 370,000 couples who still do not become pregnant are able to recognize problems early, so they are motivated to go to hospitals and receive infertility treatment after (2), resulting in early pregnancy as a result. Is done.

(1) Timing therapy is a method of estimating the date of ovulation based on the fact that the period before and after the date of ovulation is most likely to become pregnant, and adjusting the timing of sex accordingly. There are three well-known methods for estimating the date of ovulation: (A) basal body temperature measurement, (B) simple urinary LH test, and (C) follicle size measurement.

(A) is a method that has been known for a long time and is the most popular method, and is performed using a female thermometer. When menstruation begins, the low temperature period continues, transitions from the low temperature period to the high temperature period after the day of ovulation, and repeats a basal body temperature cycle with a change range of 0.3 to 0.5 ° C that returns to the low temperature period just before the next menstruation. Since the rise in basal body temperature occurs after the day of ovulation, it is too late to wait for the rise in basal body temperature and adjust the timing. For this reason, the individual's basal body temperature cycle is learned several times, and the date of ovulation is estimated based on this. However, in addition to the unstable body temperature measurement itself, the date of ovulation changes depending on the physical condition and the like, and it is said that an error of about ± 3 days is caused in the estimation of the ovulation date.

(B) is a method of capturing luteinizing hormone (Luteinizing Hormone, LH) secreted from the pituitary gland in the urine using immunochromatography, which is a simple test device. Since LH release occurs at once, it is called LH surge, and the change in concentration over time shows a mountain profile with a half width of about 24 hours and a base width of about 48 hours in blood and urine. It is known that ovulation is promoted by LH surge, and ovulation occurs in about 36 hours from the start of LH surge and in about 12 hours from LH surge peak. Here, the LH surge peak indicates the peak of the peak profile, and the LH surge indicates the peak profile itself. The urinary LH test is repeated discretely at intervals of about 24 hours from several days before the expected date of ovulation, and the date of ovulation is estimated based on the obtained signal. The principle of the immunochromatography method is as follows. The primary antibody (immobilized antibody) against the antigen to be tested (LH in the case of this test) is placed on the permeable membrane such as nitrocellulose stored inside the container of the handy strip-shaped test device in advance. When a solution containing a serum or urine sample containing an antigen and a secondary antibody (enzyme-labeled antibody) against an enzyme-labeled antigen is permeated into the osmotic membrane using capillary action, A sandwich complex of immobilized antibody-antigen-enzyme labeled antibody is formed. Here, when the enzyme substrate is permeated into the permeable membrane, a color reaction occurs based on the enzyme reaction at a predetermined position. The color reaction transits from none (negative) to present (positive) at the lower limit of detection of antigen. The presence or absence of the antigen is examined by visually identifying the presence or absence of coloration from the outside through the window of the container. This method is a qualitative measurement that does not have quantitativeness, but since a series of processing is simple and completes in about 5-30 minutes, and no special equipment is required, it is not limited to LH. It is popular as a law.

(C) is a method of directly observing the follicle by transvaginal ultrasound diagnosis. The follicle size is known to be 18-23 mm on the day before ovulation and 21-27 mm just before ovulation. In (1) timing therapy in hospitals, the combination of (B) and (C) is highly reliable and common. However, with current technology, it is difficult to perform (C) outside a hospital, so (B) is the main timing therapy that can be easily performed at home. For this purpose, LH simple test kits are sold not only for medical use but also for general use at pharmacies.

The knowledge about the relationship between the time change of LH concentration in serum and urine and the time of ovulation, and the estimation of ovulation date using the relationship has been reported in various literatures. In Non-Patent Document 3, an attempt is made to estimate the date of ovulation by LH test by conducting serum LH test and follicular ultrasonography for a large number of female subjects. While many other documents perform the same inspection at intervals of 24 hours, Non-Patent Document 3 performs the same inspection at intervals of 4 hours, and thus is one of the documents with the highest time accuracy. The LH test is a highly accurate quantitative measurement by radioimmunoassay. From these, this document is considered to provide the most accurate knowledge. The time course of serum LH concentration is roughly a Gaussian profile with a baseline of 20 μmIU / ml, a peak height of 200 μmIU / ml, and a standard deviation of about 14 hours, 27 hours and 20 minutes after the start of the LH surge, and It is concluded that ovulation occurs 10 hours after the LH surge peak time. Here, the LH surge start time is defined as the time when the serum LH concentration exceeds the threshold of 60 μmIU / ml. Based on these findings, the ovulation date can be estimated from the LH test results.

Non-patent document 4 attempts to estimate the date of ovulation by urinary LH test. It is considered that the time change of LH concentration in serum and urine is different in baseline and peak height, but the profile is almost similar and the time lag is about several hours. Non-Patent Document 4 reports that this time lag is about 2 hours. In addition, it is considered that the time change profile of the LH concentration changes depending not only on the type of sample but also on the sample pretreatment method, the quantitative measurement method of the LH concentration, the difference in the population of sample collection targets, and the like.

The knowledge about the effectiveness of ovulation day estimation using the LH simple test kit is also reported in various literatures. In Non-Patent Document 5, when a urine LH test was performed on 26 female subjects using a commercially available kit Ovuquick (Quidel), all cases were positively detected. Among these, 19 cases (73%) were positively detected in the range of 0-24 hours preceding the ovulation time, and 24 cases (92%) were positively detected in 0-48 hours. The kit has been shown to be effective in estimating ovulation dates.

In Non-Patent Document 6, five commercial kits ClearPlan Easy One-Step (Unipath Diagnostics), OvuKit Self-Test (Quidel), OvuQuick Self-Test (Quidel), Sure Step Ovulation Predicator (Applied Biotech), The urinary LH test by EZ-LH One Step Ovulation Kit (Norwell Technologies) and radioimmunoassay, which is a high-precision quantitative measurement, are compared for 11 female subjects. LH surge peak concentrations varied from 13.5 to 73.0 ～ mIU / ml, ovulation occurred within 24 hours from the peak time in 8 cases, and ovulation occurred in 24 to 48 hours from the peak time in 2 cases. On the other hand, the positive detection lower limit concentration of each kit varies in the range of 25.5-48.7 mIU / ml, the most sensitive kit ClearPlan Easy One-Step is 6 cases, the least sensitive kit EZ-LH One Step Ovulation Kit is Three cases were positively detected. In addition, a concentration range bordering between positive and negative was observed in each kit.

In Patent Literature 1, menstrually related hormones such as estrogen, progesterone, or LH are quantitatively measured at 24-hour intervals, and when it is confirmed that the quantitative concentration exceeds a preset concentration threshold, the ovulation period, ovulation It proposes a method to estimate the day, the date of menstruation, the expected date of delivery, etc. In the case of an LH test, when it is confirmed that the LH concentration quantitative value has exceeded a preset concentration threshold, or when it has been confirmed that it has passed the peak of the concentration profile, it is preset at each time point. Estimated as the day of ovulation

Also, in Patent Document 2, estrogen and progesterone are quantitatively measured at 24-hour intervals, points are given based on preset criteria for each concentration quantitative value and its rate of change, and each cumulative score is preset. When it is confirmed that the cumulative score threshold has been exceeded, a method for estimating the date of each time point as the start date and the end date of the fertile period is proposed. Furthermore, Patent Document 3 proposes a simple quantitative test kit that performs positive or negative quantitative measurement. In addition, according to Non-Patent Document 7, the peak of the estradiol concentration change profile exists about 24 hours before the peak of the LH surge, and ovulation occurs about 34 hours after the peak time of the estradiol concentration change profile. Are known.

Japanese Patent Laid-Open No. 2003-290230 Japanese National Patent Publication No. 9-506713 JP 2007-333695 A

One of the most basic solutions to the declining birthrate problem is to bring pregnancy to a couple who wants to become pregnant as soon as possible. The timing therapy that can be easily done at home without going to the hospital is based on the urinary LH simple test, but there are 400,000 couples who are worried about infertility but have not been to the hospital, It is important to disseminate to wider layers. For this purpose, it is effective to realize early and highly accurate ovulation time estimation as a timing therapy that can be easily performed at home without performing follicle size measurement that cannot be performed without going to a hospital. However, when performing ovulation time estimation within the scope of the prior art based on the relationship between time change of LH concentration and ovulation time, including simple urinary LH tests, (1) the time interval between collection of samples is coarse That is, the specimen collection time is shifted from the LH surge timing, and (2) the LH test is not quantitative, the estimation of the ovulation time is delayed or the accuracy of the estimated ovulation time is lowered.

It is impossible to repeat the LH test at a sufficiently short time interval compared to the LH surge time width (half-value width of about 24 hours), for example, at a time interval of 4 hours or less unless hospitalized. If you repeat the LH test in your daily life, a time interval of 24 hours or more is acceptable, and in fact many commercial LH simple test kits use a time interval of about 24 hours, for example, the first urine after getting up every morning Is recommended. Therefore, in the following, the time interval of LH inspection is set to 24 hours. As the knowledge obtained in advance, the interval between the LH surge start time and the ovulation time is Δtso, and the interval between the LH surge peak time and the ovulation time is Δtpo. The unit of time is hours (hours) unless otherwise specified, and rounded to the nearest decimal place. The unit of LH concentration is expressed as mIU / ml unless otherwise specified, and is rounded off after the decimal point.

First, consider the case where the LH test is quantitative. Patent document 1 corresponds. In the case of Patent Document 2, a cumulative score threshold is used instead of a simple density threshold, but since the estimation is performed based on the threshold excess time at the threshold excess time, the performance is essentially the same. A high-precision quantitative measurement device is placed at home without using a simple LH inspection kit and LH inspection is performed. This involves problems such as device cost, space for installing the device at home, and labor for operating the device. LH surge peak excess time tn (n is 1, 2, n2) when the quantitative value C (t) of LH concentration for sample collection time t = t1, t2, t3,. 3) is later than the actual LH surge peak time tp (tp ≦ tn). At this point, the LH surge peak excess time tn is set as the estimated LH surge peak time tep. (Tep = tn), the estimated ovulation time teo = tep + Δtpo can be derived based on the LH surge peak time. Here, the first specimen collection time is set to t1 = 0, and thereafter t2 = 24, t3 = 48,. The time tn when the ovulation time is estimated may be immediately before the actual ovulation time to (tn to to) or may pass the ovulation time to (to ≦ tn). Is unsuitable.

Similarly, LH concentration when LH concentration quantitative value C (t) (t = t1 (= 0), t2 (= 24), t3 (= 48), ...) exceeds the appropriate concentration threshold Ct The threshold excess time tm (m is any one of 1, 2, 3,...) Is later than the actual LH concentration threshold time tt (tt ≦ tm). At this time, by setting the LH concentration threshold excess time tm as the estimated LH surge peak time tep (tep = tm), the estimated ovulation time teo = tep + Δtpo can be derived based on the LH surge peak time. Here, when the concentration threshold is set to the concentration threshold Cs indicating the start of the LH surge (Ct = Cs), the LH surge start excess time tm is later than the actual LH surge start time ts (ts ≦ tm). By setting the LH concentration threshold excess time tm as the estimated LH surge start time tes at this time (tes = tm), the estimated ovulation time teo = tes + Δtso can be derived based on the LH surge start time. Depending on the sample collection time and the LH surge timing, the LH concentration threshold excess time tm may vary in the range of tt ≦ tm ≦ tt + 24 and ts ≦ tm ≦ ts + 24, so that tep and tes, and teo based on them, are at least 24 hours. Variation. That is, the ovulation time is estimated with a resolution that is coarser than the daily unit. This occurs because the presence or absence of the LH surge is only quantitatively determined without considering the profile of the LH surge. Even when tep and tes are not written, the same performance is obtained when ovulation time is estimated based on tn and tm.

Next, consider the case where the LH test is not quantitative. This is a case where qualitative measurement is performed using an LH simple test kit. In this case, in addition to the problem in the case where there is the above quantitative property, the following problem newly arises. The LH simple test kit has optimum sensitivity when the detection lower limit concentration Cd matches the concentration threshold Cs indicating the start of LH surge (Cd = Cs), and the ovulation time estimation based on the LH surge start time is quantitative in the LH test. The performance is equivalent to the above case. However, it is technically difficult to fix the detection lower limit concentration Cd of the LH simple test kit to a constant value, and even the same product has variations due to individual differences. In addition, since the LH concentration time change profile varies depending on the individual and the population, Cs also varies. That is, in most cases, the LH simple test kit has low sensitivity and Cd> Cs, or high sensitivity and Cd <Cs. When the sensitivity of the LH simple test kit becomes low, the time tm when the LH surge start can be recognized (the time when the positive detection is first obtained) is further delayed than when the LH test is quantitative, and the sample collection time and LH surge timing This increases the chance that LH surge peaks cannot be positively detected. If the sensitivity becomes lower and the detection lower limit concentration Cd exceeds the LH surge peak concentration Cp (Cd> Cp), the LH surge peak cannot be detected positively at all, and the ovulation time cannot be estimated. On the other hand, when the LH simple test kit becomes highly sensitive, the chance of recognizing the start of the LH surge at the baseline stage before the start of the LH surge (tm <ts) increases, and the ovulation time is estimated with no grounds. descend.

The main object of the present invention is to provide a monitoring method, a monitoring system, and a test kit for pregnancy support, which solve each of the above-mentioned problems and realize an early and highly accurate estimation of the LH surge peak time and the ovulation time. To do.

An example of a representative example of the present invention is as follows. That is, the present invention is a method for monitoring a fertile state, the step of bringing a biological sample into contact with a detection device, detecting the hormone concentration in the biological sample and storing it in a memory, and the collection time of the biological sample And the hormone concentration to be measured in the collected biological sample is detected to exceed a hormone concentration threshold value stored in advance in the memory, and the hormone stored and set in the memory is detected. Extracting the concentration time change profile function of the concentration and fitting the hormone concentration stored in the memory and the collection time, and the time at which the hormone concentration takes the maximum value from the fitting function obtained by the fitting or Estimating the time when the hormone concentration exceeds a threshold indicating the start of surge; and And time, and a time interval between pre-stored ovulation time in the memory, characterized by a step of calculating and outputting the ovulation time.

The present invention enables early and highly accurate ovulation time estimation by the LH test, and therefore enables highly reliable timing therapy that can be easily performed at home without going to the hospital. As a result, many couples who want a natural pregnancy, especially those who are worried about infertility and feel resistance to going to the hospital, have used this method, resulting in many couples becoming pregnant early, The number of births is expected to improve. In addition, for couples who do not readily become pregnant even if the present invention is used, notice early on the possibility that some kind of infertility treatment is necessary, and by prompting hospital visits, as a result, early pregnancy results, The number of births is expected to improve. In other words, the present invention is expected to save many couples who want to become pregnant and provide a solution to the problem of declining birthrate.

The figure which shows the relationship between the standard time in Nonpatent literature 3, and LH density | concentration in Example 1 of this invention. The figure which shows the area | region enclosed in the average +/- standard deviation of LH density | concentration in FIG. The figure which shows the relationship between the standard time of # 1 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 2 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 3 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 4 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 5 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 6 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 7 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 8 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 9 of Table 2, and LH density | concentration. The figure which shows the relationship between the standard time of # 10 of Table 2, and LH density | concentration. FIG. 2 is a diagram showing an approximate LH concentration time variation profile derived from Method 1 from FIG. The figure which shows the fitting result of the approximate LH density | concentration time change profile by this method 1 regarding # 7 of Table 2. FIG. 3 is a diagram showing an approximate LH concentration time variation profile derived from FIG. The figure which shows the fitting result of the approximate LH density | concentration time change profile by this method 2 regarding # 7 of Table 2. FIG. FIG. 6 is a diagram showing a performance comparison of LH surge peak time estimation between the

present methods

1 and 2 and the conventional method in Example 1 of the present invention. FIG. 6 is a diagram showing an approximate LH concentration time change profile derived by the present method 1 from the relationship between the standard time of Non-Patent Document 4 and the LH concentration in Example 2 of the present invention. FIG. 6 is a diagram showing an improvement in performance of LH surge peak time estimation by renewal or improvement of an approximate LH concentration time change profile in Example 2. The schematic diagram of the simple quantitative test kit and system in Example 3 of this invention. FIG. 5 is a schematic diagram when an optical sensor chip is used in Example 3. FIG. 4 is a schematic diagram of a configuration and chemical reaction of an inspection device in Example 3. The figure which shows the calibration curve acquisition result of hCG by the method of FIG. FIG. 4 is a schematic diagram of the configuration and chemical reaction of an inspection device and an external measurement device in Example 3. The figure which displays the LH fixed_quantity | quantitative_assay result, the LH surge peak time estimation result, and the ovulation time estimation result in Example 4 of this invention. The figure which displays the estimation result of the pregnancy possible period and the contraceptive possible period in Example 4 of this invention. The flowchart which shows an example of the estimation process at the time of ovulation of this invention.

According to the monitoring method, monitoring system, and test kit of the present invention, a quantitative LH test is performed as a timing therapy that can be easily performed at home without going to the hospital. Although the time interval of the LH inspection is arbitrary, the case of 24 hours will be described as a typical example. According to the present invention, the quantitative value C (t) (t = t1 (= 0), t2 (= 24), t3 (= 48),...) Of the LH concentration detected by a detection device or the like is input, Using the information stored in the calculation device and memory, output at least one of the estimated LH surge start time tes, estimated LH surge peak time tep, estimated ovulation time teo calculated by a unique algorithm, and early and highly accurate A method, kit or system for estimating ovulation time is provided. In particular, when the time at which the quantitative value C (t) first exceeds the concentration threshold Ct is tm (m is any of 1, 2, 3, ...), C (t1), C (t2),・・ Early estimation of ovulation time is performed by inputting only C (tm).

FIG. 27 shows an example of the flow of ovulation estimation processing according to the present invention.
In the monitoring method of the present invention, the biological sample is brought into contact with a detection device, the hormone concentration in the biological sample is detected and stored in a memory (S01 to S03), and the collection time of the biological sample is stored in the memory. And detecting that the concentration of the hormone to be measured in the collected biological sample exceeds a threshold value of the hormone concentration stored in advance in the memory, and the concentration time change of the hormone concentration stored and set in the memory. A step of extracting a profile function and fitting the hormone concentration stored in the memory and the collection time (S04 to S06), and a time at which the hormone concentration takes a maximum value from the fitting function obtained by the fitting or A step (S07, S09) of estimating a time when the hormone concentration exceeds a threshold value indicating a surge start, and the estimated time; And calculating and outputting the ovulation time from the time interval until the ovulation time stored in advance in the memory (S08, S10).

That is, the algorithm of the estimation process at the time of ovulation in the present invention is as follows. Time-discrete quantitative value of LH concentration of a certain population shown in literature etc., or time-discrete quantification of LH concentration acquired for a specific population or individual as a true LH concentration time change profile TP (T) The values are TP (T1), TP (T2), TP (T3), and so on. That is, to simplify the explanation, it is assumed that TP (T1), TP (T2), TP (T3), ... have no error from the true value. Here, T is a standard time standardized so that the LH surge peak time becomes 0, and represents only a part of one physiological cycle. For TP (T1), TP (T2), TP (T3),..., First, an approximate LH concentration time change profile AP (T) is obtained (S05). AP (T) becomes an LH surge peak at T = 0, and the peak value is Cp (AP (0) = Cp). Since the LH concentration time-varying profile represents a complex physiological phenomenon in humans and is a phenomenon that has not been solved analytically so far, there is no basis for expressing AP (T) with a specific function. Therefore, consider expressing AP (T) using, for example, a Gaussian curve, a straight line (line segment), a combination of a plurality of straight lines (line segments), a polynomial curve, and the like. Even functions other than these need only be able to approximate TP (T1), TP (T2), TP (T3), ... well.

Next, at time tm, C (t1), C (t2),..., C (tm) is subjected to fitting that translates AP (T) in the time axis direction to obtain AP (t). (S06). In general, TP (T1), TP (T2), TP (T3), ... are statistical values such as the mean value in the population, whereas C (t1), C (t2), C ( t3), ... are non-statistical values determined for each physiological cycle of each subject. At this stage, the time axis is converted from the standard time T to the sample collection time t. As a means for fitting, the least square method may be used, or AP (t) may pass a specific quantitative value, for example, C (tm). In addition to simple parallel movement in the time axis direction, fitting with correction of coefficients other than the time in the function may be performed.

Subsequently, the time when AP (t) = Cp is assumed as the estimated LH surge peak time tep (S07), and the estimated ovulation time tep + Δtpo based on the LH surge peak time is derived (S08). In the conventional method, tep = tm, whereas in this method, tep ≠ tm. In rare cases, tep ≦ tm, but in most cases, tm <tep. Moreover, the estimated ovulation time tes + Δtpo based on the LH surge start time is derived by setting the time when AP (t) = Cs (select the smaller one when there are two solutions) as the estimated LH surge start time tes (S09) ( S10).

In the conventional method, tes = tm, whereas in the estimation method according to the present invention, tes ≠ tm often occurs. Although it is rare that tm ≦ tes, in most cases, tes <tm. Since the conventional method did not consider the LH surge profile, the estimated ovulation time had a variation of at least 24 hours, whereas this method estimated the ovulation time considering the LH surge profile. The estimation accuracy is improved.

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.

As Example 1 of the present invention, an example will be described in which the estimated ovulation time teo is acquired by fixing the approximate LH concentration time change profile AP (T) based on the flow of FIG. In the following, the performance evaluation of the ovulation time estimation method according to the present invention will be described in detail with particular reference to FIGS.

Many literatures show average quantitative values TP (T1), TP (T2), TP (T3), ... of time-discrete LH concentrations in a population. The data of quantitative values C (t1), C (t2), C (t3),... Of LH concentration is not disclosed for each case in each physiological cycle. Also, since the time interval between T1, T2, T3,... Is about 24 hours, the time accuracy of TP (T) is only about 24 hours. Therefore, it is difficult to evaluate whether the method according to the present invention (the present method) is superior in the early and high-precision estimation of the ovulation time as compared with the conventional method using only the data shown in the literature.

Therefore, the appropriate LH concentration quantitative values C (#j, t1), C (#j, t2), C (#j, t3), ... are generated by simulation based on the data shown in the literature. Using these, we show that this method is superior to the conventional method for early and high-precision estimation of ovulation time. Here, #j (j = 1, 2, 3,...) Represents a unique number of a case determined for each physiological cycle of each subject. In the following, it is shown that the accuracy of the estimated LH surge peak time tep by this method is particularly excellent. The same conclusions are derived for the accuracy of the estimated LH surge start time tes and the estimated ovulation time teo based on these.

In Non-Patent Document 3, unlike many other documents, serum LH test and follicular ultrasonography are performed at 4-hour intervals, so high time accuracy of about 4 hours can be obtained for TP (T). The control data of FIG. 2 of Non-Patent Document 3 shows the mean and standard deviation of the quantitative values of serum LH concentration at 4-hour intervals. These data are read manually at 24 hour intervals (standard time T = -192, -168, ..., -24, 0, 24, ..., 168, 192), and average AV and standard deviation SD are numerical values. The results are shown in Table 1, and the graphed results are shown in FIG. 1 (average: black circle plot, standard deviation: error bar).

Where standard time is T1 = -96, T2 = -72, T3 = -48, T4 = -24, T5 = 0, T6 = 24, T7 = 48, T8 = 72, T9 = 96, T10 = 120, Define T11 = 144. Let TP (T) be the black circle plot and the solid broken line connecting them. At standard time T5 = 0, TP (T) shows an LH surge peak with a maximum value of 200 ± 89 μmIU / ml. In Non-Patent Document 3, the concentration threshold indicating the start of LH surge is set to Cs = 60 μmIU / ml. This is indicated by a dotted line in FIG. On the other hand, in FIG. 2, the average + standard deviation at the standard time Tk (k = 1, 2,..., 11) in the range of −96 ≦ T ≦ 144 in FIG. 1 (shown as AV + SD in Table 1) A broken line 1 connecting the two points and a broken line 2 connecting the mean-standard deviation (shown as AV-SD in Table 1) are shown by dotted lines. Here, at the standard time T, a function that outputs a random value in a range between the polygonal line 1 and the polygonal line 2 is RP (T).

The following procedure generates appropriate LH concentration quantitative values C (#j, t1), C (#j, t2), ..., C (#j, t10). The sample collection time is t1 = 0, t2 = 24, ..., t10 = 216. First, consider # 1. Let R (# 1) be a random value of 0 <R (# 1) ≦ 24. The standard time is set to a random value -96 + R (# 1) in the range of -96 <T ≦ −72, and the random value RP between the above broken line 1 and the broken line 2 at this standard time -96 + R (# 1) Let (−96 + R (# 1)) be the quantitative value at t1 = 0 (C (# 1, 0) = RP (−96 + R (# 1))). In addition, the random value RP (-72 + R (# 1)) at the standard time -72 + R (# 1) 24 hours after the standard time is set as the quantitative value at t2 = 24 (C (# 1, 24 ) = RP (-72 + R (# 1))). Similarly, after t3 = 48, the random value RP (T) every 24 hours is used as the quantitative value. To generalize, let R (#j) (j = 1, 2, 3, ...) be a random value where 0 <R (#j) ≤ 24, and C (#j, tk) = RP (tk-96 + R (#j)) (k = 1, 2, ..., 10). Result of generating quantitative LH concentration values at t1 = 0, t2 = 24, ..., t10 = 216 for 10 independent cases of # 1, # 2, ..., # 10 by this method Is shown in Table 2.

For each of the cases # 1, # 2, ..., # 10, the standard time T = tk-96 + R (#j) and the LH concentration quantitative value RP (tk-96 + R ( #j)) is shown as a square plot. Table 3 shows R (# 1), R (# 2),..., R (# 10) used in this case. Since the LH surge peak occurs at T = 0 at the standard time, it occurs at T = t−96 + R (#j) = 0, that is, at t = 96−R (#j) at the sample collection time. This is shown as Table 3 as LH surge peak time tp.

Here, using only the information in Table 2 (ie, under the conditions where R (#j) and tp in Table 3 are unknown), the conventional method and the algorithm according to the original method of the present invention, the LH surge peak in Table 3 is used. Estimate time tp and compare their performance. At this time, among the information in Table 2, when the time when the LH concentration quantitative value first exceeds the concentration threshold Ct is tm (m is any of 1, 2, 3, ...), t1, t2,・・ Use only tm information. The information in Table 2 is data that is randomly distributed according to the actual situation both in the time axis direction and in the concentration axis direction, so it is considered appropriate for comparing the ovulation time estimation methods.

First, the LH surge peak time is estimated by a conventional method with quantitative LH inspection. The concentration threshold value Ct of the LH concentration quantitative value is set to the concentration threshold value Cs = 60 μmIU / ml indicating the start of the LH surge recommended in Non-Patent Document 3 (Ct = 60 μmIU / ml). From Table 2, the sample collection time tm at which the LH concentration quantitative value exceeds the threshold value 60 is tm = t3 = 48 in # 7 and # 10, and tm = t4 = 72 in other cases. Also, if the concentration threshold Ct is lower than Cs and Ct = 30 mIU / ml, tm = t2 = 24 for # 7 and # 10, tm = t4 = 72 for # 8, tm = t3 otherwise = 48. On the other hand, if the concentration threshold Ct is higher than Cs and Ct = 120 mIU / ml, the LH concentration quantitative value does not exceed the threshold in # 1, # 7, and # 8, and in # 2, tm = t5 = 96, otherwise tm = t4 = 72. As described above, the estimated LH surge peak time is set to tep = tm at the sample collection time tm in each case except when the LH concentration quantitative value does not exceed the threshold value. If the LH concentration quantitative value does not exceed the threshold value, the LH surge peak time cannot be estimated. The results are summarized in Table 4.

In Table 3, the correct LH surge peak time tp is recorded, and at the same time, the deviation from each estimated LH surge peak time tep. When estimation is impossible, a cross is indicated in the column. Moreover, the average AV of # 1 to # 10 of each item, the standard deviation SD, and the number of x are shown. It can be seen that the lower the concentration threshold, the earlier the estimation can be performed, but the deviation increases. Conversely, the higher the concentration threshold, the smaller the deviation, but the estimation execution is delayed and the estimation cannot be performed. Therefore, among these, the concentration threshold Ct = 60 μmIU / ml is considered to be most suitable for the early and highly accurate estimation of the LH surge peak time. On the other hand, in the conventional method that does not have quantitativeness like the LH simple test kit, the concentration threshold Ct is often unknown or the concentration threshold Ct is often deviated from the optimum value 60. Even if the LH simple test kit is the same product, the concentration threshold Ct varies depending on individual differences. For example, if the concentration threshold Ct varies between 30 and 120, tep varies between the threshold 30 AV and SD in Table 4 and the threshold 120 AV and SD, so the LH surge peak time estimation performance is quantitative. It is inferior to some conventional methods.

Next, the LH surge peak time tep is estimated according to the present invention. First, the approximate LH concentration time change profile AP (T) is expressed by a Gaussian curve. Fitting a Gaussian curve to all the black circle plots in Fig. 1 under the condition of T = 0 and peak value Cp = 200 mIU / ml (AP (0) = 200), AP (T) = 177 * exp (T / 19) +23 is derived. Here, when AP (T) is derived, AP (0) = 200 does not have to be a condition. This AP (T) is shown by a solid line in FIG. 13 together with the black circle plot and error bar in FIG. The concentration threshold value Ct of the LH concentration quantitative value is set to the concentration threshold value Cs = 60 μmIU / ml indicating the start of the LH surge recommended in Non-Patent Document 3 (Ct = 60 μmIU / ml). Subsequently, for each of # 1 to # 10, t1, t2,..., の tm (tm is the sample collection time that first exceeds the concentration threshold 60) and its LH concentration quantitative value C (t2), C (t2),..., C (tm) is fitted to AP) (T) only by translation in the time axis direction by the method of least squares. For example, in the case of # 7, tm = t3, and AP (t) = 177 * exp ((t−69) / 19) +23 is derived.

In Fig. 14, the time axis is converted from the standard time T to the sample collection time t, the data of # 7 in Table 2 is indicated by a rhombus plot, and AP (t) is indicated by a solid line. Add arrows to the first three square plots used for fitting. In addition, a correct LH surge peak time tp = 75 is indicated by a dotted line. The LH surge peak time is estimated at the sample collection time tm = t3 = 48. The estimated LH surge peak time tep is the peak time of AP (t), and tep = 69. The deviation -6 between the peak time tep and the correct peak time tp = 75 indicated by the dotted line is the estimation error. The results of similar estimations for all cases # 1 to # 10 are summarized in Table 5 as “Method 1”. As in Table 4, the number of deviations, tp, AV, SD, and x are also shown. Based on the estimated LH surge peak time tep, the estimated ovulation time teo is derived by tep + Δtpo. According to TABLE 3 of Non-Patent Document 3, Δtpo = 10 ± 5, so teo = 79 ± 5.

Next, the approximate LH concentration time change profile AP (T) is expressed by a line segment. Of the straight lines passing through the two black dot plots of T = −24 and T = 0 in FIG. 1, a line segment satisfying the condition of 0 ≦ AP (T) ≦ 200 is defined as AP (T). AP (T) = 5.67 * T + 200 (0 ≦ AP (T) ≦ 200), which is indicated by a solid line in FIG. 15 together with the black circle plot and error bar in FIG. The concentration threshold value of the LH concentration quantitative value is Ct = 60 as before. Subsequently, for each of # 1 to # 10, AP (T) is passed so as to pass through tm in Table 2 (tm is the sample collection time that first exceeds the concentration threshold 60) and its LH concentration quantitative value C (tm). Fitting is performed only by parallel movement in the time axis direction. For example, in the case of # 7, tm = t3, and AP (t) = 5.67 * (t−71) +200 (0 ≦ AP (T) ≦ 200) is derived. In FIG. 16, the time axis is converted from the standard time T to the sample collection time t, the data of # 7 in Table 2 is indicated by a rhombus plot, and AP (t) is indicated by a solid line. Add an arrow to the third square plot used for fitting. In addition, a correct LH surge peak time tp = 75 is indicated by a dotted line. The LH surge peak time is estimated at the sample collection time tm = t3 = 48. The estimated LH surge peak time tep is the time at the upper right end of the line segment of AP (t), and tep = 71. The deviation -4 between the peak time tep and the correct peak time tp = 75 indicated by the dotted line is the estimation error. The results of similar estimations for all cases # 1 to # 10 are summarized in Table 5 as “Method 2”. As in Table 4, the number of deviations, tp, AV, SD, and x are also shown.

比較 Comparing AV and SD of Method 1 and Method 2, it can be said that the performance of LH surge peak time estimation is almost equivalent. Comparing Table 4 and Table 5, in most cases, the deviation of this method was smaller than that of the conventional method. Only in # 3 and # 10, the deviation of the conventional method (threshold 120) was smaller than the deviation of this method. This is because, in the conventional method (threshold 120), the correct LH surge peak time tp and the execution time tm of LH surge peak time estimation are close to each other. Comparing AV and SD, it can be seen that Method 1 and Method 2 have better LH surge peak time estimation performance than either conventional method.

Furthermore, in order to statistically clarify the performance comparison evaluations in Tables 4 and 5 above, the number of cases was increased from 10 to 100 from # 1 to # 10, and the same analysis was performed. Table 6 and Table 7 show the results of summarizing the numbers of SD, SD, and X, respectively. FIG. 17 shows the frequency distribution of 100 deviations according to each method. The upper part of Fig. 17 shows the conventional method (threshold 30) as a black triangle plot, the conventional method (threshold 60) as a black circle plot, and the conventional method (threshold 120) as a black square plot. The lower part of Fig. 17 shows this method 1 as a round plot. This method 2 is indicated by a white triangle plot.

From Table 6, the upper part of FIG. 17, in the conventional method with quantitativeness in the LH test, the deviation and its variation decrease as the threshold value increases, but there is a problem that the estimation is impossible. For this reason, the threshold 60 is optimal, and the LH surge peak time can be estimated with an accuracy of −14 ± 12 hours 14 hours before the average. Here, if the average deviation -14 is known in advance, the average deviation can be zeroed by setting tep = tm + 14 without setting the estimated LH surge peak time at tep = tm. The value of 14 is a value determined by TP (T), Ct, and the time interval of sample collection, and can be derived by the above analysis.

That is, this method is another means for solving the problems of the present invention. However, this method does not change the standard deviation of the deviation, so the performance is lower than Method 1 and Method 2. In addition, in the conventional method in which the LH inspection does not have quantification, it is difficult to apply this method because the threshold value varies.

In contrast, from Table 7 and the lower part of Fig. 17, Method 1 and Method 2 have almost the same performance, and the deviation and its variation are greatly reduced compared to the conventional method (threshold value 60). . In Method 1, the LH surge peak time can be estimated with an accuracy of −1 ± 7 hours before an average of 14 hours. Further, in this case as well, the average of the deviations can be made zero by setting the estimated LH surge peak time to tep ′ = tep + 1.

Next, Example 2 will be described. In Example 1, the approximate LH concentration time variation profile AP (T) was fixed and the performance evaluation of ovulation time estimation was performed.In this example, AP (based on the LH test results of individual physiological cycles of individual subjects The performance of ovulation time estimation is improved by renewing or improving T).

Non-Patent Document 4 attempts to estimate the date of ovulation by LH test by conducting urinary LH test and follicular ultrasonography on a large number of female subjects. Non-patent document 4 Table 7 shows the urinary LH concentration in 24-hour intervals from the 10th day to the 20th day, with the first day of the menstrual cycle as the first day in the case of one menstrual cycle of 7 subjects. The quantitative results are shown. For the seven cases, the day when the LH concentration is maximum is set to the standard time T = 0, and ± 1 day, ± 2 days,... Based on this day, T = ± 24, ± 48,. Set to. The average and standard deviation of the LH concentration quantitative values in seven cases at each standard time are shown by a black circle plot and an error bar in FIG. 18, respectively. However, there is no error bar for T = -144, -120 because there is only one target case. In addition, when a Gaussian curve is fitted to all the black circle plots in FIG. 18 under the condition of taking a peak value at T = 0, the approximate LH concentration time change profile AP (T) = 60 * exp (T / 17) +14 is derived.

Compared to AP (T) = 177 * exp (T / 19) +23 derived for Non-Patent Document 3 in Example 1, the peak width of the Gaussian curve does not change much, that is, the profile has the same shape, The peak height is about 1/3 and the baseline is about 2/3. The main cause is considered to be that the sample has changed from serum to urine, but there are other differences in conditions such as the sample pretreatment method, the LH concentration quantitative measurement method, and the sample collection target population. Example 1 is a comparison between the conventional method and the present method with the conditions fixed without considering these differences, but the comparison result is considered to be effective under other conditions. Therefore, when estimating the ovulation time by this method 1 for LH concentration quantitative value using urine as a specimen, AP (T) = 60 * exp (T / 17) +14 is at least AP (T) = Higher estimation accuracy can be expected than using 177 * exp (T / 19) +23. Conversely, when AP (T) = 60 * exp (T / 17) +14 was used to estimate the ovulation time according to this method 1 for the LH concentration quantitative value using serum as a sample, it was compared with Example 1. Then, how much the estimation accuracy decreases will be evaluated next.

Estimate ovulation time using this method 1 for 5 cases # 6 to # 10 shown in Table 2 using AP (T) = 60 * exp (T / 17) + 14 for the LH concentration quantitative value . Here, the density threshold value is set by (peak height) * 20% + (baseline) (Cs = 177 * 20% + 23≈60) following the case of the first embodiment. In other words, Cs = 60 * 20% + 14 = 26 is set here. The average ± standard deviation of the deviations in each case was calculated to be −29 ± 9. As shown in Non-Patent Document 4 on the leftmost side of FIG. 19, the average is plotted on the top and the standard deviation is plotted on the bottom. In addition, for the above five cases, when ovulation time estimation was performed according to Method 1 using AP (T) = 177 * exp (T / 19) +23 and Cs = 60 in Example 1, −2 ± Similarly, a black circle is plotted as Non-Patent Document 3 on the rightmost side of FIG. This is a result equivalent to −1 ± 7 shown in Table 7. That is, it can be seen that the ovulation time estimation accuracy is significantly lowered by improperly setting the approximate LH concentration time change profile. On the other hand, it is next evaluated how much the ovulation time estimation accuracy can be improved by renewing or improving AP (T) based on the history of individual LH test results.

In Table 2, # 1, # 2, ..., # 5 are the LH concentration quantitative values for the first, 2, ..., 5 physiological cycles of the same individual, and # 6 to # 10 are the LH concentrations of the same individual. Assuming a quantitative value. First, it is assumed that the first physiological cycle is over and only the LH concentration quantitative value of # 1 is obtained. When a Gaussian curve is fitted to all the rhombus plots of FIG. 3 which are quantitative LH concentration values of # 1, AP (T) = 80 * exp (T / 39) +22 is derived. At this time, the approximate LH concentration time change profile is updated from AP (T) = 60 * exp (T / 17) +14 to AP (T) = 80 * exp (T / 39) +22. The density threshold is Cs = 80 * 20% + 22 = 38 according to the same rule as before. In order to perform an accurate comparative evaluation, when the ovulation time was estimated by this method 1 for 5 cases # 6 to # 10 using this AP (T) and Cs, the deviation was 5 ± 8. Plot black circles as # 1 data second from the left. Compared to Non-Patent Document 4, there is a significant improvement in ovulation time estimation. Next, it is assumed that the second physiological cycle is over and only the LH concentration quantitative values of # 1 and # 2 are obtained. When the average value of the LH concentration quantitative values of # 1 and # 2 is obtained at each standard time, and a Gaussian curve is fitted to them by the least square method, AP (T) = 124 * exp (T / 24) +25 Is derived. At this point, the approximate LH concentration time change profile is improved from AP (T) = 80 * exp (T / 39) +22 to AP (T) = 124 * exp (T / 24) +25. The density threshold is Cs = 124 * 20% + 25 = 50. When these are used to estimate the ovulation time in the same manner as described above, the deviation is −2 ± 8, and the black circle is plotted as # 1- # 2 data third from the left in FIG. Similarly, the result of estimating the ovulation time of # 6 to # 10 by obtaining the approximate LH concentration time change profile of the Gaussian curve from the average value of # 1 to # 3, # 1 to # 4, and # 1 to # 5 These are shown in FIG. Table 8 summarizes AP (T) and Cs used in each ovulation time estimation of FIG.

From the above results, it can be seen that it is very effective to upgrade or improve AP (T) based on the history of individual LH test results. AP (T) created with just one LH concentration quantitative value for only one physiological cycle is also effective, and by using LH concentration quantitative values for two or more physiological cycles, it is almost ideal approximate LH concentration time The change profile AP (T) can be derived. However, it is assumed that the variation between physiological cycles of the LH concentration time change profile TP (T) of the same individual is relatively small. On the contrary, if this condition is satisfied, the decrease in ovulation time estimation accuracy due to differences in specimen type, specimen pretreatment method, LH concentration quantitative measurement method, specimen collection target population, etc. should be solved by the above means. Can do.

The method of renewing and improving AP (T) shown in this embodiment is an example, and the same effect can be obtained by other methods.

In order to perform ovulation time estimation according to the present invention, LH concentration quantification means including time measurement means is required. Although an immunoassay apparatus for clinical tests may be used as a high-precision quantitative measurement method, it is not realistic in terms of apparatus cost and installation space to perform daily tests at home. In recent years, instead of visually reading the results of various test kits using immunochromatography, devices that automatically read optically and quantify the substances to be tested have been developed by Roche Diagnostics Co., Ltd., Otsuka Electronics ( Etc.) are sold for in-situ inspection (POCT). Light intensity of the color reaction by irradiating the color reaction part on the nitrocellulose film from the outside through the window of the container of the immunochromatography test kit with a light emitting diode or laser diode, and measuring the reflected or transmitted light. Is measured. Although it is possible to implement the present invention using an equivalent device, although it is a desktop device, it is still a problem that it is large and expensive for use in a general household. On the other hand, handy and low-cost devices that optically read the color reaction of immunochromatography are sold by Inverness Medical Japan Co., Ltd. for LH testing and from Arax Co., Ltd. for hCG testing (pregnancy testing). However, these are qualitative measurements that digitally determine positive or negative and cannot be quantitatively measured. On the other hand, the simple quantitative inspection kit proposed in Patent Document 3 solves the above-described problems of quantitative accuracy, device size, and device cost, and is suitable for implementation of the present invention. In the following, the effect of the present invention is further enhanced by improving the kit of Patent Document 3.

FIG. 20 shows a schematic diagram of the simple quantitative inspection kit and system of the present invention. An external measurement device such as the reader 202 or the personal computer 203 includes at

least memories

205 and 208 and

calculation units

206 and 209, and the results calculated by the method of the present invention using the inspection device 201 are displayed on the

display units

204 and 207. FIG. 21 is a schematic diagram of the case where an optical sensor chip is used, in which the color reaction of the immunochromatography method is switched to a luminescent reaction, and an RFID equipped with a photosensor in the luminescent reaction part on the permeable membrane inside the container of the inspection device 201 A chip (light sensor chip) is installed and the emission intensity is measured. FIG. 22 schematically shows the flow of the substance to be inspected, the flow and localization of various reagents, and the luminescence reaction on the permeable membrane in the inspection device 201. In this embodiment, nitrocellulose is used for the osmotic membrane, but other materials may be used. On the right end of the osmotic membrane in the dry state, the immobilized antibody is immobilized in the test part that becomes the luminescence reaction part, the photosensor chip is arranged at this position, and the photosensor chip is also arranged in the blank part as a reference in the vicinity thereof. . In addition, the enzyme-labeled antibody is held in a dry state in the specimen absorption part (urine absorption part) on the left end side of the osmosis membrane. Here, when the sample solution (urine) is immersed in the sample absorption part, the antigen (LH), which is the test substance in the sample, binds to the enzyme-labeled antibody, and at the same time, through the osmotic membrane toward the right end by capillary action. To penetrate. Since the antigen also binds to the immobilized antibody, a sandwich complex of the immobilized antibody-antigen-enzyme labeled antibody is formed in the test part. The enzyme-labeled antibody that does not form a sandwich complex passes through the test part and is absorbed by the solution absorption part. Next, when the enzyme substrate solution is immersed in the specimen absorption part, it penetrates through the osmotic membrane toward the right end and reaches the test part, where a luminescence reaction occurs with the enzyme in the sandwich complex. Luminescence intensity measurement has a wider linear response range with respect to the concentration of the test target substance than that of color intensity measurement, and therefore has excellent sensitivity and dynamic range for the determination of the test target substance.

FIG. 23 shows the results of a calibration curve obtained by quantitative measurement of hCG, which is a pregnancy test marker, instead of LH as the test object by this method. The horizontal axis represents the hCG concentration, and the vertical axis represents the emission intensity. The light emission intensity is obtained by integrating the output difference between the test part and the blank part for a certain period of time. Plots and error bars (displayed only on the upper side) show the mean and standard deviation of n = 5, respectively. In the conventional immunochromatography method based on visual judgment, the hCG concentration of 10 ⁰ to 10 ¹ ng / ml is a transition region from negative to positive, and the detection lower limit concentration exists in this concentration range. In contrast, in this method, a lower detection limit concentration of hCG concentration of 10 ^-1 ng / ml or less was obtained, and a calibration curve was obtained in a three-digit concentration range of hCG concentrations of 10 ^-1 to 10 ² ng / ml. It has been shown that quantification with high detection sensitivity and a wide dynamic range is possible. Another advantage of using the luminescent reaction is that the inspection result does not remain in the inspection device, so that even if the inspection device is discarded, the risk that the inspection result is known to others is low, and privacy protection is excellent. The inspection device cannot visually confirm luminescence from the outside, and the luminescence reaction attenuates in a short time as the luminescent substrate is depleted. Further, since the emission intensity data is not stored in the optical sensor chip, there is no fear of being read by others.

As shown in FIG. 21 (A), when the cap is connected after immersing the user's urine in the urine absorbing part of the handy size testing device 211, the enzyme-labeled antibody and the luminescent substrate previously held in the urine absorbing part and the cap are removed. It penetrates the osmotic membrane. At this time, the inspection device has a light shielding structure, and adverse effects due to ambient light can be avoided. There is no window for viewing from the outside of the container of the inspection device. In addition, since the inspection device is in a sealed state, it is possible to prevent the attached urine from being contaminated to the outside. Since the RFID chip (photosensor chip) is supplied with driving power wirelessly by a reader 212 outside the inspection device as shown in FIG. 21B, the inspection device and the optical sensor chip do not have a power source such as a battery. Therefore, the inspection device can be made disposable by suppressing the unit price by mass production of the optical sensor chip and reducing the manufacturing cost of the inspection device.

Next, the inspection device 211 is connected to the reader 212 as shown in FIG. The reader 212 is equipped with a battery as a power source. The reader is not disposable, but is used for multiple inspections. The reader 212 wirelessly feeds and controls the optical sensor chip 211 and reads the optical sensor chip ID and emission intensity data. That is, the inspection device and the reader are not connected by wire. This also contributes to a reduction in the manufacturing cost of the inspection device and at the same time improves the connection reliability. In order to easily determine the relative position between the inspection device and the reader in a predetermined relationship to ensure wireless connection, it is desirable that the reader has a socket structure of the inspection device as shown in FIG. Conversely, there is no worry that the data transfer here will be read by others from a distance. By setting the reader to a handy size, the connected inspection device and the reader can be stored in a small bag such as a cosmetic pouch. This is effective for conducting inspections at places other than home, such as toilets at work. The emission intensity data is stored in a memory mounted on the reader.

Next, as shown in FIG. 21 (C), when the emission intensity measurement is completed and a predetermined lamp on the reader informs that, the predetermined switch on the reader is pressed to transmit the emission intensity data to the external measuring device, here the user. Wirelessly transferred to the mobile phone 213 at once. About time information, it is good to memorize | store as time detected by the reader | leader side, and to make it transmit information. For this wireless transfer, any means such as infrared communication, ZigBee, etc. may be used, but safety is ensured so that it cannot be read by others. The mobile phone is preinstalled with LH concentration analysis software, ovulation time estimation software, and display software, and data necessary for analysis by the software is stored in the memory. The LH concentration analysis software quantifies the LH concentration by analyzing the luminescence intensity data, and the ovulation time estimation software estimates the LH surge start time, LH surge peak time, and ovulation time from the LH concentration quantitative value using a unique algorithm, and the display software Displays the LH concentration quantitative value, estimated ovulation time, etc. on the display as numerical values, tables, graphs, etc. In addition, the mobile phone has a function of storing emission intensity data and subsequent processing data. In recent years, most people already own and carry their own mobile phones, so by using the mobile phone as an information processing device for computing and displaying results, the simple quantitative test kit and the entire system can be substantially In addition, the cost can be reduced and the size can be reduced.

In addition, when the inspection device and reader in the connected state in FIG. 21 (B) are left in the toilet at home or stored in a cosmetic pouch, the emission intensity measurement is performed, and the user shifts to other work. It becomes possible to return to FIG. 21 (C) later when convenient. The information processing apparatus may be a personal computer or a portable information terminal as shown in FIG. 20, or a dedicated information processing apparatus. Further, the connection between the reader and the information processing apparatus may be wired, and the reader and the information processing apparatus may be separate devices as shown in FIG. 20, or may be an integrated structure device. In either case, only the inspection device is disposable for each inspection, and it is desirable that the reader and the information processing apparatus are repeatedly used for a plurality of inspections.

On the other hand, instead of incorporating the optical sensor in the inspection device as in the example of the simple quantitative inspection kit shown in FIG. 20, the external measuring device (202, 203) may incorporate the optical sensor as shown in FIG. good. In this case, a window is provided in the container of the inspection device so that the luminescence reaction can be measured from the outside of the container. If the window is closed with a transparent material such as glass, the sealed state of the container can be maintained.

Following the connection between the inspection device and reader in FIG. 21 (B), connecting the inspection device and the external measurement device to determine the relative position in a predetermined relationship, the optical sensor of the external measurement device is connected to the container as shown in FIG. It becomes possible to measure the luminescence reaction in the test part inside the container close to the window. However, in FIG. 24, the external measuring device shows only a part near the optical sensor. In addition to the optical sensor, the external measuring device includes at least a circuit for measuring emission intensity, a storage element for storing emission intensity data and time, and a battery for a power source for driving these. Therefore, the element of the optical sensor does not need to have a wireless communication function. Further, the optical sensor can be used repeatedly without being disposable. On the other hand, since the inspection device does not need to incorporate an optical sensor chip, it can be manufactured at a lower cost and is more suitable for disposable use. The information processing apparatus that performs calculation and result display may be built in the external measurement device, or may be connected to the external storage device wirelessly or by wire.

As a fourth embodiment of the present invention, means for displaying the estimation result will be described with reference to FIGS. In Example 4, on the display of an information processing device such as a mobile phone, personal computer, or portable information terminal, the LH concentration quantification result, LH surge start time, LH surge peak time, ovulation time, sex timing suitable for pregnancy ( Estimated results such as the period of pregnancy) and the timing of sex suitable for contraception (contraception period) are displayed.

Here, the case of # 7 in Example 1 is taken as an example. Repeat the LH test every 24 hours at 7am after waking up. We assume 7:00 am on Wednesday, October 15, 2008 as t1 = 0 in Table 2, that is, the date and time of the LH test start. FIG. 25 (A) shows an example of the result display of the LH concentration quantitative value on the third day from the start of the LH test. Show the bar graph with the quantitative values of LH concentration on the 1st and 2nd days so that the transition of the LH concentration can be seen at a glance. By changing the color of the LH concentration quantitative value on the third day, it is easy to understand that it is the latest data at the present time. The horizontal axis indicates the number of days counted from the start of the LH test, but may indicate the number of days counted from the start of menstruation or a date such as “10/15”. On the display, “October 17th (Friday) at 7:00 am, sample collection day: 3rd day, LH concentration: 72 (mIU / ml)” will be shown together so that the current quantitative LH concentration can be determined. To do. The results may be displayed in a graph or table other than the bar graph. Also, since the LH concentration quantitative value on the third day exceeds the concentration threshold value 60, the message “LH concentration exceeds the threshold value. Do you want to estimate the ovulation time?” Is displayed and the user is asked whether execution is possible.

When the user selects the estimation execution of FIG. 25 (A), the LH surge peak time is estimated according to Method 1 of Example 1 and the result is shown as FIG. 25 (B). The estimation may be performed according to the method 2 of the first embodiment and other algorithms. The LH concentration quantitative value is shown as a rhombus plot, and for the third day, the color of the plot is changed to make it easy to understand that it is the latest data at the present time. The horizontal axis shows the number of days from the start of the LH test, and each day starts at 7:00 am. The result of fitting a Gaussian distribution, which is an approximate LH concentration time change profile, is displayed, and the peak time is displayed as “estimated LH surge peak time: October 18 (Sat) 4 am”. This time is 69 hours after 7 am (Wednesday), October 15, 2008 (Wednesday), when the LH test starts, and is consistent with the results in Table 5. Here, it is expressed as “estimate LH surge peak 21 hours from now”, “estimate LH surge peak within 24 hours from now”, “estimate LH surge peak 12 to 24 hours from now”. It is also effective to clarify the comparison with the current time of Friday, October 17, 2008 at 7:00 am. When the error of the estimated LH surge peak time can be expected, for example, if the error is ± 7 hours, “estimated LH surge peak time: Saturday, October 18 4 ± 7 am” or “estimated LH surge peak time: 10 May 17 (Friday) from 9:00 pm to October 18 (Saturday) at 11:00 am ”. Further, as shown in the first embodiment, the LH surge start time may be estimated and the result may be displayed.

Next, 10 hours after the estimated LH surge peak time is estimated as the ovulation time according to Example 1, and “estimated ovulation time: October 18 (Saturday) 2 pm” is displayed as shown in FIG. To do. Here, it is expressed as “estimate ovulation 31 hours from now”, “estimate ovulation within 36 hours from now”, “estimate ovulation 24 to 36 hours from now”, and the current time of 2008 It is also effective to clarify the comparison with 7:00 am on Friday, October 17th. If an error in the estimated ovulation time based on the estimated LH surge peak time can be expected, for example, if the error is ± 5 hours as shown in Non-Patent Document 3, the probability distribution of the ovulation time is a Gaussian with a standard deviation of 5 hours. Since the distribution can be assumed, the probability distribution is displayed so as to overlap with the fitting result of the approximate LH concentration time change profile as shown in FIG. The two Gaussian distributions are color-coded, or one Gaussian distribution region is colored so that they can be easily distinguished from each other. Also, as shown in Fig. 25 (D), based on an error of ± 5 hours, "estimated ovulation time: October 18 (Saturday) 9 am-7 pm" is displayed, and this time region is approximated by LH You may display it superimposed on the fitting result of the density time change profile. Estimated ovulation time: Only the time of 2:00 pm on Saturday, October 18 may be displayed with a fitting result of the approximate LH concentration time change profile by a straight line or an arrow.

Subsequently, based on the estimated ovulation time, the pregnancy possible period and the contraceptive possible period are displayed as shown in FIG. For example, assuming that the sperm survival time is 48 hours and the egg survival time is 12 hours, "Estimated ovulation time: Saturday, October 18 from 9:00 am to 7:00 pm" ) “Possible pregnancy period: October 16 (Thursday) from 9:00 am to October 19 (Sunday) at 7:00 am”, and this time domain is the fitting result of the approximate LH concentration time change profile and It is displayed overlapping the time region of the estimated ovulation time. By displaying the time range of the pregnancy possible period and the estimated ovulation time in different colors, the two can be easily distinguished. On the other hand, it is assumed that the period other than the pregnancy possible period is a contraceptive period, and as shown in Fig. 26 (B), "Contraception possible period:-October 16 (Thursday) 9:00 am, October 19 (Sunday) am 7 o'clock ”is displayed, and this time region is displayed so as to overlap with the fitting result of the approximate LH concentration time change profile and the time region of the estimated ovulation time. The contraceptive period and the estimated ovulation time are displayed in different colors so that they can be easily distinguished from each other. Moreover, timing therapy can be effectively performed by displaying advice more specifically according to the user's needs.

By displaying as described above, not only early and highly accurate ovulation time estimation can be obtained, but also a sense of security and satisfaction can be obtained when performing timing therapy, and there is less chance of getting lost in timing. In addition, the display is visual and easy to understand, and you can perform timing therapy while having fun.

In the above, an example was given assuming that the LH inspection is repeated every 24 hours, but the time interval need not be limited to this, and the time interval need not be constant in a series of LH inspections. The method of the present invention can be applied to either case.

In the above embodiment, the ovulation time is estimated before ovulation for the purpose of estimating the ovulation time early and with high accuracy, but also using the results of the LH test after ovulation, Therefore, the ovulation time may be estimated by the method of the present invention. Since the latter case uses a larger amount of information for estimation, the accuracy of ovulation time estimation is further improved. This allows you to look back on the past after ovulation, check the transition, evaluate your own judgment and behavior, and improve the probability of pregnancy after the next time by reflecting it in the subsequent judgment and behavior. Become. In addition, displaying the history of the user's LH test results for each menstrual cycle or displaying the history of a plurality of organizing cycles in an overlapping manner can be useful for timing determinations from the next time onward.

In the above examples, the LH surge peak time or LH surge start time is estimated from the LH concentration quantitative value, and the ovulation time is estimated based on these times, but the method of the present invention is used for hormones other than LH. May be. For example, estradiol (estrogen) is a hormone that is secreted by many women just before ovulation, just like LH. According to Non-Patent Document 7, it is known that the peak of estradiol concentration change profile exists about 24 hours before the peak of LH surge, and ovulation occurs about 34 hours after the peak time of estradiol concentration change profile. ing. That is, if the peak time of the concentration change profile is estimated from the estradiol concentration quantitative value by this method and the ovulation time is estimated based on this time, the ovulation time can be estimated earlier than in the case of LH. . Further, by applying the present invention to a plurality of hormones, such as LH and estradiol, and integrating the estimation of ovulation time performed for each of them, a more accurate estimated ovulation time can be derived. It is easy for the user to display the concentration change profiles and estimated ovulation times of a plurality of hormones or to display the integrated estimated ovulation times in an overlapping manner. As a means for integration, the sum of probability distributions of ovulation times obtained for each may be used, or a product may be used. On the other hand, an estimation method of ovulation time using basal body temperature may be displayed together with this method. For example, if the changes in basal body temperature and changes in the LH concentration quantitative value for each day are overlaid, and the ovulation time estimated from each or the integrated estimated ovulation time is overlaid, more accurate ovulation time estimation is possible. Become.

Furthermore, other effects can be obtained by sharing information with the specific person through the network safely through the estimation result obtained above. For example, by sharing information with a partner, it becomes easier to obtain cooperation from the partner, and the success probability of timing therapy can be improved.

The present invention provides a test kit and system for hormones such as LH that realizes early and highly accurate ovulation time estimation.

201, 211: Inspection device, 202, 212: Reader, 203, 213: External measuring device, 204, 207: Display, 205, 208: Memory, 206, 209: Calculation unit.

Claims

A method for monitoring a fertility state,
Contacting a biological sample with a detection device, detecting a hormone concentration in the biological sample and storing it in a memory;
Storing the collection time of the biological sample in the memory;
Detects that the concentration of hormone to be measured in the collected biological sample exceeds a threshold value of hormone concentration stored in advance in the memory, and extracts a concentration time change profile function of the hormone concentration stored and set in the memory And fitting the hormone concentration and the collection time stored in the memory;
From the fitting function obtained by fitting, a step of estimating the time when the hormone concentration takes a maximum value or the time when the hormone concentration exceeds a threshold indicating the start of surge;
And a step of calculating and outputting the ovulation time from the estimated time and a time interval until the ovulation time stored in the memory in advance.
2. The monitoring method according to claim 1, wherein the fitting is an application of a least square method to the hormone concentration and collection date / time data stored in the memory of the concentration time change profile function. How to monitor.
3. The monitoring method according to claim 1, wherein the fitting is parallel movement of the concentration time change profile function in a time axis direction.
2. The monitoring method according to claim 1, wherein the concentration time change profile function is a Gaussian curve.
2. The monitoring method according to claim 1, wherein the concentration profile function is a straight line.
2. The monitoring method according to claim 1, wherein the concentration time change profile function is created by periodic data stored in the memory.
The monitoring method according to claim 6, wherein the concentration time change profile function is derived from data obtained by integrating data of a subject from whom the biological sample is collected and data not derived from the subject. A monitoring method characterized by comprising:
2. The monitoring method according to claim 1, wherein the concentration of the hormone to be measured is one or more of an LH concentration, an estradiol concentration, and an hCG concentration.
2. The monitoring method according to claim 1, wherein the biological sample is urine or blood.
2. The monitoring method according to claim 1, wherein the time at which the hormone concentration takes a maximum value or the time at which the hormone concentration exceeds a threshold value indicating the start of surge is estimated together with the deviation time stored in the memory. A monitoring method characterized by the above.
A system for monitoring a fertility status,
A detection device for contacting a biological sample to detect a hormone concentration in the biological sample and a collection time of the biological sample;
Memory,
A calculation unit;
An output unit that outputs a result calculated by the calculation unit;
The calculation unit detects that the measurement target hormone concentration of the biological sample exceeds a hormone concentration threshold value stored in advance in the memory, and the concentration time change profile function of the hormone concentration stored and set in the memory From the fitting function obtained by fitting the hormone concentration stored in the memory and the collection time, and the fitting function obtained by fitting, a threshold value at which the hormone concentration takes a maximum value or a threshold at which the hormone concentration indicates the start of surge is obtained. An ovulation time is estimated by estimating an exceeding time, and the calculated ovulation time is output to the output unit.
12. The monitor system according to claim 11, wherein the output unit outputs that the hormone concentration has exceeded a threshold value, and receives an input as to whether or not the calculation unit estimates the ovulation time. And monitor system.
12. The monitor system according to claim 11, wherein the output unit displays the estimated ovulation time together with the concentration time change profile function.
A test kit for testing a fertility state,
A reaction device for causing a reaction for measuring a measurement target substance in the biological sample by contacting the biological sample;
A detector for detecting a reaction amount of the reaction device;
A memory for storing the reaction amount and time detected by the detector, and an external calculation means including a calculation device,
The calculation device uses the reaction amount detected by the detector as a hormone concentration as the measurement target substance, detects that the measurement target hormone concentration exceeds a hormone concentration threshold value stored in advance in the memory, Extracting the concentration time change profile function of the hormone concentration stored and set in the memory, fitting the hormone concentration and the collection time stored in the memory, and fitting the hormone concentration from the fitting function obtained by fitting A test kit characterized in that the time at which the maximum value is taken or the time when the hormone concentration exceeds a threshold value indicating the start of surge is estimated to calculate the ovulation time.
15. The inspection kit according to claim 14, wherein the reaction is a luminescence reaction, the detector is an optical sensor chip, and the optical sensor chip is mounted on the reaction device. kit.
15. The inspection kit according to claim 14, wherein the optical sensor chip and the external calculation means are wirelessly communicated.
15. The inspection kit according to claim 14, wherein the external calculation means is a mobile phone.