MX2010004156A

MX2010004156A - A diagnostic device for identifying rupture of membrane during pregnancy.

Info

Publication number: MX2010004156A
Application number: MX2010004156A
Authority: MX
Inventors: Jacob Mullerad; Hagit Frenkel-Mullerad; Ronen Nahary
Original assignee: Momtec Life Ltd
Priority date: 2007-10-18
Filing date: 2008-10-22
Publication date: 2011-04-13
Also published as: AU2008313294A1; EP2198299A4; EP2198299A2; WO2009050711A2; BRPI0818779A2; WO2009050711A3; CA2703254A1; KR20100098606A; JP2011501810A

Abstract

A diagnostic device is provided for distinguishing between amniotic fluid and urine in female secretion. The device can be employed as a panty shield or can be adhered onto a panty shield or can be employed in a simple pad that is pressed against a substrate provided with female secretion.

Description

A DIAGNOSTIC DEVICE TO IDENTIFY IDE RUPTURE THE MEMBRANE DURING PREGNANCY j The present invention deals with methods, and diagnostic devices. More particularly, | the present invention deals with tests and diagnostic devices I to identify membrane rupture during pregnancy.

I BACKGROUND OF THE INVENTION Labor is different for each woman, and it can be difficult to pinpoint when it starts. In jvez to be a unique event, involves a whole process, with several changes and physiological events that occur in the body and combine to then give birth to a baby. One of those changes or events is the rupture of the membrane that surrounds the baby in the uterus, which is a clear I indication that labor is imminent. Sometimes, rupture of the membrane occurs abruptly and obviously, with an abundant flow of amniotic fluid. However, on other occasions, a pregnant woman may have a broken membrane and be insecure or unconscious of whether the membrane is broken or not. i There are two main reasons for this. First, I The baby's head can act as a plug in the entrance I of the uterus and in that way, instead of the amniotic fluid spurting out, it comes out very slowly.

? In addition, the amniotic fluid may leak out through a small opening in the uterus, and the pregnant woman may not feel the first contractions until a few hours later. Secondly, during late pregnancy, women tend to have difficulty controlling themselves when urinating. Because of this, the continuous flow of amniotic fluid from the vagina will not be noticed as the woman is accustomed to small amounts of urine escaping from time to time.

The wrong and untimely diagnosis of amniotic fluid spill, especially during pregnancies of high risk, may result in a failure to implement > an appropriate treatment, which increases the risk in pregnant women and their fetus. The risks | of neonatal complications due to spillage of inadvertent amniotic fluid include acute fetal distress, infections and premature birth and can cause very dangerous consequences for both the mother and the fetus.

Currently, the only available way for a pregnant woman to detect amniotic fluid output at home is through a self-diagnostic diagnostic pads kit, which is very expensive. Therefore, almost all pregnant women choose to go to their doctors or to the emergency room of a hospital. In the hospital, the tests currently available for the i Amniotic fluid identification are invasive, inaccurate or expensive and in all cases can cause discomfort for the patient. There is a great need to reduce the possibility of the membrane breaking without complaining at home by using a precise device, Quick and cheap that can also act as a tool for the doctor to perform a test in the clinic, or hospital.

Previous Technique j Some examples are described below! of several conventional methods currently available to physicians. The most used method for the analysis) of urinary compound is by means of the well-known technique chorno "strips of test of reaction and reading" or "reactive strips". The test strips are test strips made of reactive ageJtes attached to a reagent-bearing matrix (cushion) on a strip. In general, the strip is made of polymeric materials such as polyethylene, polycarbonate or polystyrene. Each carrier matrix contains a different reactive agent. The test strip should be impregnated with a urine sample and removed. After the reagents in the carrier matrix and urine come into contact, a color change reaction occurs. A test strip can be designed either as a test strip with a single cushion (for the test of a single analyte) or as a test strip with multiple cushions (to test several different analytes at the same time). The test strips can be used manually or with a suitable chemical analyzer. A test strip with multiple profiles is described for performing multiple simultaneous or sequential analyzes of the analytes in the U.S. Patent numbers. 4,595,439, 4,526,753, 4,160,008, 3,123,443, 3,212,855, 3,814,668, 4,038,485, 3,531,254.

Several of said test devices 1 are available in the market. The following is a partial list of brands of test strips CLINISTIX, ULTISTIX, KETOSTIX, N-MULTISTIX, DIASTIXK, DEXTROSTIX, AUTION STICKS, CHEMSTRIP.

The reactive carrier matrix is usually an absorbent material that allows the liquid sample; move through the matrix. This movement of the liquid sample is in response to the capillary forces that; they are formed in the matrix. During the movement of the liquid through the matrix, it makes contact with the chemical reactive compound impregnated in the matrix. Therefore, the color transition that can be detected and measured occurs. If the test strip can measure several analytes simultaneously, the color change in each reagent carrier matrix can be related to the different amount or analyte in each liquid sample. The manual analysis of the results requires the comparison of the developed color of the Test on the test strip with a table of colors. , The material of the reagent carrier matrix can be any substance that can incorporate the chemical reagents necessary to carry out the test (of interest.) The preferred matrix must be inert with respect to the chemical reagents and must not alter the sample of the results. The reagent carrier matrices can be made of several materials, some of these materials are: papers containing fibers such as filter papers, woven and non-woven fabrics, modified synthetic and natural polymers, spongy materials, cellulose, fiber glass, micropore membranes, and wood The reactive mat may also be different with respect to roughness and smoothness along with softness and hardness.

The following list of patents describes the use of different matrices: US. Pat. Nos. 3,846,247, 3,552,928, 3,902,842, 3,418,083.

Sewell DL, and associates discloses, among other things, the cost of using the test strip as a screening method for urinalysis in a scientific paper published in the American Journal of Clinical Pathology, Vol. 83; (6) pages. 740-743, 1985. The authors stipulate that the use of the dipstick procedure "costs approximately $ 0.76 per reagent". Several devices are described in the text to determine particular urinary analytes with the use of reactive carrier matrices (filter paper, micro capsules, test strips, etc.). The following list of test devices used by prior art techniques includes dry tablets, test strips, and other techniques for the analysis of urinary constituents. The U.S. patent do not. 4, 147, 514 describes the detection of ketone bodies; Pat. U.S. do not. 3, 146, 070 discloses dry chemical compounds in a carrier (dipstick) impregnated with a pH indicator for pH determination. Method, compound and test devices are disclosed to determine the ionic strength or specific gravity of a sample; of test like urine in the following U.S. patents 4,318,709, 5,403,744.

The Jaffe method is a well-known method for the determination of creatinine. This method involves the formation of an orange-red color with a solution of alkaline picrate. Pat. U.S. do not. 6,001, 656 discloses a device for a creatinine test in liquid test samples 1. The improvement in this patent involves the inclusion of one or more selected quinolines in the reactive formula. Another method for the determination of creatinine; is the one described by Benedict and Behre in the Journal of Biological Chemistry (1936), which involves the reaction of 3, 5-dinitrobenzoic acid with creatinine in an alkaline medium. HE disclose other methods, compounds, and proving devices for determining creatinine in a liquid sample such as urine in the following U.S. Patents. 4,215,197 (using; an enzymatic compound), 5,662,867, 5,733,787. Appropriate materials for the detection of creatinine include picric acid, 3, 5-dinitrobenzoic acid, 3,4-dinitrobenzoic acid, 2,4-dinitrobenzene sulphonic acid, t (3, 5-dinitrobenzo) yl alcohol, (3,5-dinitrobenzo) -nitrile, (3, 5-dinitrobenz) amide and N, N-diethyl- (3, js-dinitrobenz) amide. j There have been reports of other methods for the determination of proteins in fluids. These methods include the Biuret method, the Lowry method, the method í Kjeldahl, the method of dye combination, fluorometric method and UV method. Of these methods, the Kingbury-Clark method is widely used; presented in J. Lab. Clin. Med., 11, 981 (1926) and the Meulemans method; presented in Clin. Chem. Acta, 5, 757 (1960) and the brilliant blue method of Coomassie; informed in An l. Biochem. 72, 248 (1967). The Bradford dye test for protein determination, Pat. U.S. do not. 4,023,933 is also routinely used in almost every biochemical laboratory. In general, the protein interacts with substances, mainly with dyes such as Coomassie bright zul, bromophenol blue (blue tetrabromophenol), and eosin as well as metal iohes such as copper (II), lead (II) zinc (II) and silver (I). The addition of protein-containing solution to the reaction between the dye and a metal ion provided a I Spectral change to a metal ion-dye solution. j Other protein indicators include those described as well as merocyanine and substituted polyhalogenated substituted nitro or nitroso phenolsulfonphthalein disclosed in U.S. Pat. No. 5,279,790. Other protein indicators are Fast Green FCF, Light Green SF, Red Pyrogallol and I pyrocatechol violet, bromochlorophenol blue (3 ', 3": dibromo-5', 5" - dichlorophenolsulfonphthalein), basic fuchsin, basic violet, martius yellow, phloxin B, yellow ! methyl, Congo red, orange methyl and ethyl orange (4- (4-diethylaminophenylazo) benzenesulfonic acid). The following list of U.S. patents it has to do with the measure j of proteins in solution, as is the urine, using reactive systems usable in a dipstick method: 5,424,215, 5,593,895, 6,815,210, 4,960,710, 3,485,5j87, 5,087,575, 4,023,933.

Several test strips used for urine test j contain tests for urobilinogen. The classic urobilinogen test, developed by Paul Ehrilich in 1901, uses paradimethylaminobenzaldehyde as a test for which, in a strong acid medium produces a coffee-orange color - i i I red with Ehrlich reagent. For more information on the urobilinogen, the Ehrlich reaction and the urobilinogen j test are described in Tietz, Textbook, de i Clinical Chemistry, W. B. Saunders Company. Some examples of U.S. patents and test strips for determination! of urobilinogen based on the Ehrlich reaction and j the diazonium coupling reaction are: Pat. U.S. us. i 3,853,466, 3,630,680, 4,665,038, 4,290,771, 3,989,462, 3,814,586. More U.S. patents containing information on the measurement of urobilinogen in solution, such as urine, using reactive systems usable in the dipstick method are: 4,158,546 and 3,447,905. ! The oldest method of the alkaline phosphatrasease test was introduced in 1930 by Kay. Then, a popular test method for the determination of alkaline phosphatase using p-nitrophenyl phosphate was introduced in 1946 by Bessey, Lowry and Brock. This method depends on the fact that after being exposed to the fluids containing alkaline phosphatase, the colorless p-nitrophenyl phosphatase is catalytically hydrolyzed in a p-nitrophenol product of yellow chlorine (and phosphate). Therefore, the concentration of! the enzyme is determined by following the increased intensity | of the yellow color of the reaction product. The activity: alkaline phosphatase occurs naturally in natural milk, where after pasteurization, j the I enzyme is denatured. In this way, the activity of alkaline phosphatase is used in an indicator for the proper pasteurization of milk. One such test: dry alkaline phosphatase activity in milk is the PHOSPAHTESMO MI test, manufactured by MACHEREY - NAGEL GmbH & Co.

In the presence of hemoglobin in the urine is called hemoglobinuria, this condition can occur as a result of a red cell lysis (RBS) in the urinary tract. The term hematuria is used when intact RBS are present in the urine. This condition can occur in bleeding from the renal or genitourinary systems. The most used tests for, the detection of blood in the urine or in the feces depends on the fact that the hemoproteins act as peroxidases. This reaction requires a hydrogen donor molecule. Typical examples of products that include tests for blood detection are ULTISTIX 10SG, HEMOCCULT II, and AUTION STICKS.

More recently, the United States Patent 4,357,945, published on November 9, 1982 to Janeo for the DEVICE FOR TESTING AND BREAKING AMNIOTIC MEMBRANE, describes a manual coupling device with a pH indicator. When exposed to fluids, the indicator changes color if the amniotic membrane broke. The U.S. Patent 5,425,377, issued June 20, 1995 to Caillouette by pH MEASUREMENT OF BODY FLUID, discloses a wet structure on a stick, provided with a pH indicator for measuring the pH of the vaginal fluid.

Various other methods are provided in the following: U.S. Patent 5,554,504 published September 10, 1996 to Rutanen by DIAGNOSTIC METHOD FOR DETECTING FETAL MEMBRANE BREAKDOWN, which describes the detection of factor binding protein, of growth of insulin type 1 in a vaginal discharge sample. U.S. Patent 5,281,522, published January 25, 1994 to Senyei and associates as REAGENTS AND KITS FOR THE DETERMINATION OF FETAL FIBRONECTIN IN A VAGINAL SAMPLE, describes kits for the detection of rupture of membranes by the sample of a cavity. vaginal and by exposing it to antibodies such as the anti-fetal fibronectin antibody and an anti-fibronectin antibody. U.S. Patent 5,096,830, published on March 17, 1992, to Senyei and associates as PREMATURE BIRTH AND TEST OF MEMBRANE BREAKDOWN describes a method to determine: rupture of the fetal membrane by removing a sample from the vaginal cavity and placing it in contact with an insoluble support to which the ani-fetal antigen antibody adhered, and the binding of the fetal antigen to the support is determined.

Several devices are known that have to do i with pantyhose or tampons with indicators of ??. ' Below are examples of some of these patents: U.S. Patent 6,149,590 published Nov. 21.

From 2000 to Smith and company as SYSTEM TO IDENTIFY PREMATURE BREAKING OF THE MEMBRANE DURING PREGNANCY, describe a cushion that has an upper external layer, a lower external layer, and an intermediate compound that reacts to the pH. U.S. Patents 6,921,647 and 6,627,394 published to Kritzman and associates as SECRETION MONITORING ARTICLE and DIAGNOSTIC CUSHION describe cushions | with pH-sensitive indicators for spill detection.

I amniotic fluid. U.S. Patent 5,217,444 published to Schoenfeld as ABSORBENT TAMPON demonstrates an absorbent material containing a pH indicator material which indicates by a color change the acidity! or alkalinity of a liquid that comes into contact with it.

It should be mentioned that the tests that show only the pH of the vaginal discharge for the differentiation of the ! Amniotic fluid in the urine is not accurate because of the following: 1) The pH of the urine can vary from 4.5 to 8 depending on the homeostatic activity of the kidneys and the intake of water, and 2) the pH of the amniotic fluid varies, from 6. 9 to 7.15 in late pregnancy. This superimposed range of i pH can lead to a false diagnosis that can cause medical complications. j i ? Immunochromatographic tests that are based on the detection of antibodies with specific proteins in; Amniotic fluid or urine, are not commonly available to the public especially because it has a price ajlto because they contain monoclonal / police antibodies.

In general, enzymatic methods are also expensive for i the high cost of enzymatic production.

Summary of the present invention It is an object of the present invention to provide unique devices and methods to differentiate amniotic fluid from urine based on the analysis of more than one analyte using dry chemical reactions without the need I to use enzymatic or immunological methods.

It is another object of the present invention to provide unique devices and methods for differentiating amniotic fluid from urine that are simple and that save time and money, which in the end helps resolve the dilemma of whether labor will begin soon or not. Once the i first drop of spilled liquid has contact with; the device, the woman or health care personnel will be able to observe immediately if said fluid contains amniotic fluid or urine. I Still another object of the present invention! is to provide unique devices and methods to differentiate amniotic fluid from urine that can be used with strands ? pregnant women either human or not. The determination of the onset of labor in non-human females may be even more unpredictable due to the inability of the animal to communicate its condition.

In addition, another object of the present invention is the point of care practice of the rapid chemical analysis of biological fluids such as urine, saliva, sweat, cerebrospinal fluid (CSF), milk or liquids from source otaras. I Therefore, it is provided according to a preferred embodiment of the present invention i dry diagnosis device to distinguish between amniotic fluid i and urine in female secretion, where the dry diagnostic device consists of: A base layer; At least two inert carrier matrices í provided in said base layer; | Dry reagents provided in said inert carrier matrices wherein said dry reagents are capable of forming a chemical reaction with substances in the female secretion so as to distinguish visually the difference between amniotic fluid and urine where said I dry reagents in each of at least two carrier matrices is capable of reacting with substances other than said substances. i In addition, and in accordance with another preferred embodiment of the present invention, said dry reagents are capable of reacting with substances present in | the amniotic fluid or in the urine, where the substances have a much higher concentration in one of the two liquids than in the other. j In addition, and in accordance with yet another preferred embodiment form j of the present invention, said substances include substances such as creatinine, alkaline phosphatase, total protein, urea, urobilinogen and blood.

In addition and according to yet another preferred embodiment of the present invention, one of the dry reagents in one of at least two carrier matrices is capable of reacting with the creatinine.

In addition and in accordance with still another preferred embodiment of the present invention, one of said carrier matrices consists of two reactive layers, one of which contains creatinine-sensitive dye fixed with a dye-fixing agent and a second one. layer that i contains a shock absorber capable of maintaining said zone! test at a relatively high pH value. j In addition, according to yet another form; of preferred embodiment of the present invention, one of the aforementioned carrier matrices consists of a single layer which contains a creatinine sensitive dye, a buffer, and a dye fixative agent. ! In addition, and according to yet another preferred embodiment of the present invention, said creatinine-sensitive dye is selected from a group of dinitro derivatives such as 3 '5'-dinitrobenzamide acid, 2'4'-dinitrobenzoic acid, '5' - dinitrobenzotrifluoride, 3 '5'-dinitrobenzamide, 3' 5'-dinitrobenzoyl phenyl glycine, 3,5-dinitrohydroxyphenylpropionic acid.

In addition and according to yet another form of i preferred embodiment of the present invention, said dye fixing agent selected from polymerized quaternary ammonium cations (quats) such as polydiallyldimethylammonium chloride, polyimonoalyltrimethylammonium chloride, polytrimethylaminoethyl methacrylate chloride, polyvinylbenzyltrimethylammonium chloride, polyvinyl methylpyridine chloride i.

In addition and according to still another preferred embodiment 1 of the present invention, said creatinine sensitive dye and said dye fixing agent are buffered so as to maintain a stable pH in the range of about 9 to 13.5.

In addition and in accordance with yet another preferred embodiment 1 of the present invention, it is selected a buffer of a group of sodium metasilicate, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydroxide - potassium chloride, potassium carbonate, glycine - sodium hydroxide, and sodium borate.

In addition and according to yet another preferred embodiment of the present invention, one of the dry reagents in one of the carrier matrices is capable of reacting with a total protein.

In addition and according to yet another preferred embodiment of the present invention, said dry reagent is a dye selected from a group of 3 ', 3", 5', 5" - tetrabromophenolsulfonphthalein, Coomassie brilliant blue, Fast Green, Light Green, Pirogaliolsulfonphthalein (red pyrogallol), pyrocatecholsulfonphthalein (pyrocatechol violet), 3 ', 3"dibromo-5', 5" - dichlorophenolsulfonphthalein, fuchsin acid, 2,4-dinitro-1-naphthol (yellow martius) floxin B, Congo red, ethyl orange and methyl orange.

In addition and according to yet another form! of preferred embodiment of the present invention, the device further comprises a buffer such as potassium citrate, potassium chloride, potassium sulfate, iodate | of potassium or potassium phosphate.

In addition and according to yet another form! of preferred embodiment of the present invention, the I The device also consists of a metal ion selected from copper, lead, zinc, silver. i In addition and according to yet another form | from In the preferred embodiment of the present invention, one of the dry reactants in one of the carrier matrices is capable of reacting with the alkaline phosphatase. i In addition and according to still another preferred embodiment of the present invention, said dry reagent is an alkaline phosphatase substrate selected from p-nitrophenyl phosphate, indoxyl phosphate, 4-methylumbelliferyl phosphate and alpha-naphthyl phosphate. ! In addition and in accordance with still another preferred embodiment of the present invention, one of said dry reagents is a pH sensitive reagent.

In addition and in accordance with yet another preferred embodiment of the present invention, said reagent can not be removed from the carrier matrices. 1 Furthermore, and in accordance with still another form of the preferred embodiment of the present invention, at least two of said carrier matrices are covered by a protective layer.

In addition and according to yet another form | of preferred embodiment of the present invention, said protective layer is transparent. I In addition and according to yet another form, of Preferred embodiment of the present invention, said protective layer may be made of a thin "single path structure" membrane permeable to liquids flowing to said carrier matrices and prevents the flow of reagents out of the device.

In addition and in accordance with still another preferred embodiment of the present invention, a sticky reinforcing layer is provided below the aforesaid base layer.

In addition and in accordance with still another preferred embodiment of the present invention, said sticky reinforcing layer is provided adjacently with an outer protective layer.

In addition and in accordance with yet another preferred embodiment of the present invention, said reagent carrying matrices are arranged in substantially parallel lines.

In addition and according to yet another preferred embodiment of the present invention, said reagent carrying matrices are arranged in concentric lines.

In addition and in accordance with yet another form; of preferred embodiment of the present invention, said reactive carrier matrices are surrounded by adhesive material which allows adhesion of the device of diagnosis when said at least two test zones oppose the vaginal canal of a female animal.

Brief description of the illustrations Some embodiments of the invention are described herein, only as an example, with reference to the accompanying drawings. With specific reference to the detailed illustrations, it is emphasized that the features shown are examples only and are used only as illustrative discussion method of the preferred embodiments of the present invention, and are presented with the objective of providing what the most understandable and useful description of the principles is believed | and conceptual aspects of the invention. With respect to this, it is not intended to show structural details of the invention in more detail than is necessary for the fundamental understanding of the invention. The description obtained from the drawings will allow those skilled in the art to see how to use the invention in practice in various ways.

In the drawings:! Figure 1 illustrates a cross-sectional view of the device: diagnostic according to a preferred embodiment of the present invention.

Figures 2 to 5 illustrate diagnostic devices attached to women's panties according to the I preferred embodiment of the present i I invention. ! i Figures 6 a-d Illustrate diagnostic cushions according to the preferred embodiment of the present invention.

Figure 7 illustrates a veterinary diagnostic cushion! according to the preferred embodiment of the present invention. ! Detailed description of the invention ' Before explaining in detail at least one embodiment of the invention, it should be understood that the invention is not necessarily limited in this respect to the I details set forth in the following description 1 or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

The terms "consist of", "compounds", "includes", "including", and "having" together with their conjugates mean "including but not limited to" | The term "consisting of" is equivalent | in meaning to "including and limited to". 1 The term "consisting essentially of" means that the compound, method or structure may include ingredients, steps and / or additional parts, but only if the ingredients, steps and / or additional parts do not alter! in a material way the basic and novel characteristics of the compound, method or structure in question.

As used herein, the singular form "a," "an," and "the" includes plural references unless the context clearly dictates otherwise. For example, the terms "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

Through this application, various embodiments of this invention can be presented. It should be understood that the description in the range format is for convenience and brevity only and should not be considered as an inflexible limitation in the field of the invention. According to the above, the description of a range should be considered as a specific disclosure of all possible sub - ranges as well as the individual numerical values within the range.

It is appreciated that certain features of the invention, which are described, as a clarification, in the context of separate embodiments, may also be provided in combination in a single embodiment. Likewise, various features of: the invention, which are, for brevity, described in the context of a single embodiment, may be provided separately or in any possible sub combination or as deemed appropriate in any other described embodiment of the invention. Some characteristics described in the context of various embodiments are not considered as essential characteristics of those embodiments, unless the embodiment is useless without those elements.

The present invention provides unique and novel devices for distinguishing amniotic fluid from urine in a simple and rapid way to allow a pregnant woman to know her pregnancy condition. In accordance with one aspect of the present invention, a device that resembles an 'adhesive tape', which is a disposable strip containing sensitive chemical indicators, is used during the last months of pregnancy. The strip is relatively small in dimensions and consists of at least two layers, an adhesive reinforcement and an absorbent material containing the indicators. The woman who uses the device only has to adhere to her panties with the test area facing the body. In case the woman wore a pantiprotector, you can place the strip on top of it. Rapid chemical reactions in the test areas of the device can cause a distinctive color change in the event of amniotic fluid leakage and there is a distinct change in color when the urine comes in contact with the test zones. The device of the present invention acts as a diagnostic tool for allow the user to detect if the secretion contains amniotic fluid or only urine, according to a known color index which is provided in the device. j I In accordance with the second aspect of the present invention, a device which resembles a cushion and which contains sensitive indicators independently and in contact with the female body is used. This device can be used when pressing it against wet underwear or! a wet pantiprotector after noticing the fluid spill. This will cause the reagents in the reagent-carrying matrices to come into contact with body fluids and almost immediately after the spilled liquid comes in contact with the diagnostic device, chemical reactions occur in the reagent-carrying matrices and cause a change of color. Again, the woman (or caregiver), using this device, will be able to see whether the fluid contains amniotic fluid or just urine, according to the known color index that is provided in I the device.

According to the third aspect of the present invention, a device similar to a pantiprotector contains sensitive indicators that allow the distinction between amniotic fluid and urine. The pregnant woman will stick this product to her underwear and continue with her daily life. The indicator reactive areas should Position themselves opposite the vagina. In this way, 1 the sensitive pantiprotector will be in close contact with the woman's bodily fluids.

In all aspects of the invention, if: post-collection storage is required, the sample can be transferred to a suitable container for storage. Alternatively, the immediate process of the sample can be performed. If used, the sample should be placed directly on the device's reactive carrier matrices and the test performed within a few minutes after sample collection.

According to another aspect of the present invention, a veterinary device is provided. The device consists of sensitive reagents that will be directly attached to the vaginal opening of the animal female. This can be achieved in a simple manner by means of a design of the vda where the adhesive zone is around or on the sides of the test zone of the reactive carrier matrices. Therefore, close contact with the body fluids of the animal will be achieved. You can also achieve the use of! a device without direct contact with the animal body by obtaining the liquid sample with a swab having a fibrous tip or by means of a suction or washing device, and by applying it to the indicator areas of the devices. I I According to a preferred embodiment of the present invention, the determination of whether there exists the presence of amniotic fluid or urine in the secretion of the woman relapses into several separate non-enzymatic and non-immunological reactions that can be determined as associated either with the urine or the amniotic fluid through its distinctive color. In most of the tests currently available for the identification of the cause of humidity during pregnancy, only the difference in the concentration of an analyte in the body is measured.

I or in the amniotic fluid. In the present invention,! HE I identifies the existence of at least two of the following substances: protein, creatinine, urea, urobilinogen, blood, and alkaline phosphatase, as well as pH. Basically, it was identified that protein and alkaline phosphatase j are present in high concentrations in the liquid ! amniotic compared to urine while creatinine, urea and urobilinogen occur in higher concentrations in urine compared to amniotic fluid. Blood may exist in the amniotic fluid in greater concentration than in normal urine. The chemicals that can be used to distinguish between liquids according to a preferred embodiment of the present invention are those which when bound or reacted with one of the substances indicated herein or actuate due to certain medium they change in spectroscopic properties.

Therefore, the diagnostic device consisting of reagent carrying matrices having chemical indicators according to the present invention is capable of detecting at least two of the following substances: protein, creatinine, urea, urobilinogen, blood, and alkaline phosphatase , as well as the pH value. The method of the present invention is based on the following factors: 1) The concentration of total protein in the amniotic fluid is normally 15 times higher than its concentration in the urine, 2) The concentration of creatinine and urobilinogen in the urine it is normally about 10 times more than its concentration in the amniotic fluid, and 3) The concentration of urea in the urine is usually more than 2 times its concentration in the amniotic fluid, 4) The concentration of alkaline phosphatase in the amniotic fluid is usually more than 7 times greater than the concentration in the urine, 5) during the rupture of the fetal membranes! the amniotic fluid that comes out may contain blood, which is contrary to the urine of a healthy woman. Therefore, even if a small amount of amniotic fluid is present in the vaginal secretion and comes into contact with a cushion of the present invention, the cushion allows diagnosis of fetal membrane rupture with extremely high precision. You can also detect oras i substances with similar concentration differences.

The present invention is also explained by the following examples of devices provided with sensitive indicators for detecting and distinguishing the spill of urine or amniotic fluid.

Reference is made to Figure 1 showing a diagnostic device according to a shape; of preferred embodiment of the present invention, in a top view and transverse view, respectively. 1 A device 10 has at least two, and preferably several reagent carrying matrices 12. Figure 1 shows a device that adapts to stick to the woman's body and is therefore covered with a first layer 14 of soft material and comfortable that does not irritate the skin on contact so that it can be used on women's panties. The first layer 14 avoids direct contact between the reagents that are provided in the device 10 and the adjacent skin.

It should be noted that the first layer 14 can be transparent to allow easy visual distinction of the colors formed in the layers below it.

The device 10 further comprises a support base layer 16 which may contain the carrier matrices; of reagent 12 capable of carrying out the tests of interest. The reagent carrier matrices 12 are preferably! an absorbent material that allows the liquid sample to be move through the matrix. The reagent carrier matrices 12 must be inert with respect to the chemical reagents and must not alter the sample or the results of the test.

Optionally, the carrier matrices of reagents 12 can be made of materials, for example: papers containing fibers such as filter papers, woven and non-woven fabrics, modified synthetic or natural polymers, sponge materials, cellulose, glass fiber, micropore membranes, and wood. Some additional materials may be microporous polymeric materials such as styrene-based polymer, latex-based, cellulose-based, or cotton-based matrices.

Reagent carrier matrices can also have different characteristics such as roughness, smoothness, softness and hardness.

It should be noted that in the manufacturing process, the reagent carrier matrices can be made to have several layers, some of which can carry different reagents in different areas of the test zone. Any combination of the support base layer and the additional matrices that carry the indicators are found I covered by the field of the present invention and for no reason is intended to limit the scope of the present invention.1 Under the support base layer 16, i it provides a sticky reinforcing layer 18 and also an outer adjacent protective layer 20. Those two layers are similar in nature with the layers that are provided in the protection of the panty-protectors. j The reagent carrier matrices 12 are They are preferably arranged in groups where each group has indicators capable of indicating one of the substances that were listed herein. Below are the examples of chemical reagents and methods and preparation of each one's test zone: Examples ^ I Preparation of the creatinine test zone I The concentration of creatinine in the urine is usually 10 times greater than the concentration of the amniotic fluid. The dry test of creatinine is done in two optional ways: ' • A system of two reactive layers that contain! a fixed dye sensitive to creatinine that is placed in a matrix and placed in the second a buffer that causes the system to maintain a value of; Relatively high pH j • A reactive layer system containing | a creatinine-sensitive dye, a buffer !, and fixative agents, all placed in a matrix. The creatinine sensitive dye can be one i i I of the dinitro derivatives like:! 3, 5-dinitrobenzoic acid, 2,4-dinitrobenzoic acid, 3,5-dinitrobenzotrifluoride acid, 3,5-dinitrobenzamide, 3,5-dinitrobenzoyl-phenyl glycine, 3,5-j-dinitrohydroxyphenyl propionic acid.

The sensitive dye fixative agent includes | from 1 preference polymerized quaternary ammonium cations i (quarts) such as: polydiallyldimethylammonium chloride (Poly j DADMAC), polymonoalyltrimethylammoniumchloride polytrimethylaminoethyl methacrylate chloride, and polyvinylbenzyltrimethylammonium chloride, chloride; of polyvinyl tilpyridine. ^ The shock absorber must be a strong base to keep the pH stable in a range of between 9 and 13. 5 . Examples of such buffers are sodium metasilicate, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydroxide-potassium chloride, potassium carbonate, glycine-sodium hydroxide, and sodium borate. . I Optionally, the non-volatile solid reagents 1 are added to the sensitive dye in order to improve I the reaction. j Two-layer reactive system: Example 1 The creatinine test indicator consisting of two reagent carrier matrices was installed in the qapa i ! base of the diagnostic device. The first matrix (can be a Whatman filter paper or napkin paper) | it was impregnated with creatinine sensitive dye, 3 'j 5' - dinitrobenzoic acid (dBA), and poly DADMAC (optional) as a fixing agent which dissolved in water. The concentrated solution of dBA was mixed with the sodium carbonate buffer. A second reagent matrix was impregnated with sodium metasilicate buffer. After both reagent matrices were dried, the first reagent matrix j was placed on the second reagent matrix, which was impregnated with the sodium metasilicate buffer.

Both dry reagent matrices were placed in layer b se of the device. I i After a drop of vaginal secretions comes into contact with the reagent carrier matrix, a distinctive color change will identify the fluid and indicate whether it contains amniotic fluid or urine. j Two-layer reagent system: Example 2 j Two absorbent reaction matrices were each impregnated with a solution, dried and introduced into! the base layer of the device. The first reaction matrix) J impregnated with a solution 1 consisting of dBA (concentrated solution that was dissolved in Sodium Hydroxide) as a creatinine-sensitive dye, of solid reagent, not I volatile and poly DADMAC (optional) as a fixing agent.! The The second reagent matrix was impregnated with a 2 g solution including sodium hydroxide. Both reaction matrices j were dried. The dry reagent matrix with solution 1 was placed on the dry reagent matrix with solution 2 and both were introduced into the base layer of the device.

Reagent layer system: Example 1 j As mentioned before, "one layer: reagent system" has the same rational as the "two reagent layer" system but contains all the chemicals in a reagent carrier matrix. In both cases, the objective) is the differentiation between urine and amniotic fluid using the concentration of creatinine. j The creatinine test device was prepared from the same absorbent and support carriers as in the two-layer reagent system. The reagent carrier matrix consists of a creatinine sensitive dye, a non-volatile solid reagent, a fixative agent (optional) and! a damper that was dried and installed in the carrier matrix. There was a preferable use of 3 '5'-dinitrobenzoic acid (the concentrated solution was made in acetonitrile) as a sensitive dye, the poly DADMAC (optional) as a fixing agent (optional) and Potassium Hydroxide (in alcohol solution). ethyl) as a buffer. The reagents were dried together and readied for the test of a urine sample or amniotic fluid. i System of a reagent layer; Example 2 An emulsifed solution of 3 '5'-dinitrobenzoic acid, poly DADMAC (optional), acrylic styrene acid, sodium metasilicate and a solid, non-volatile reagent was prepared. A thin layer of the emulsion was spread on a latex-based matrix, and dried. The dried matrices were introduced into the polymer of the support base layer of the device. ! Preparation of the Total Protein test zone The concentration of the total protein in the amniotic fluid is usually 15 times greater than its concentration in the urine. The interaction of proteins and substances, mainly dyes and metal ions, causes a spectral change in the metal ionic solution -color. When implementing the device of the present invention, a group of dyes and metal ions such as: 3 ', 3", 5', 5" tetrabromophenolsulfonphthalein, Coomassie bright blue, Fast Green, Light Green, Pirogaliolsulfonphthalein (red pyrogallol) can be used. ), pyrocatecholsulfonphthalein (pyrocatechol violet), 3 ', 3"- dibromo - 5', 5" - dichlorophenolsulfonphthalein, fuchsin acid, 2,4 - dinitro - 1 - naphthol (yellow martius) Copper, lead, zinc, silver, phloxine B, Congo red, ethyl orange and methyl orange.

The reagent carrier matrix of the device Total protein test is an absorbent carrier which can be one of the aforementioned matrices, for example the microporous polymer material such as styrene-based copolymer, latex-based, cellulose-based or cotton-based matrices. The reagent carrier matrix can be a polymerized urethane-based compounds (as described in U.S. Patent No. 5, 124, 266) incorporating an indicator reagent compound capable of interacting with proteins to produce a visually detectable response.

In order to reduce the variability due to differences in the density of the urine, the test solution may include a buffer with a potassium salt base with low pH such as potassium citrate, potassium chloride, potassium sulfate. , potassium iodate or potassium phosphate.

Total protein test: Example The total protein solution test consists of a combination of two solutions: the dye reagent solution and the buffer. For the dye solution, 3 ', 3", 5', 5" - tetrabromophenolsulfonphthalein was used and dissolved in a weak organic acid such as citric acid.i The buffer, potassium citrate, was impregnated with the same weak organic acid. at a low pH value of 3.5.

The coloring solution was then diluted with. the buffer solution on a scale of large proportions. After its dissolution, a very thin layer of The resulting solution was scattered in a carrier matrix! of reagent and dried. Once the urine and amniotic fluid comes in contact with the reagents impregnated in the device, a distinctive color reaction can make the difference between these liquids. i Preparation of the pH test zone As mentioned before, the preparation is placed in a carrier matrix of absorbent reagent of any type. The reagent matrix is impregnated with a pH indicator for measuring the pH of the fluid. The pH of the urine can vary from 4.5 to 8 while the pH of the vatic amniotic fluid between 6.9 and 7.15 in late pregnancy. This range of! pH can be verified with an indicator or two different pH indicators; a low pH indicator and a medium pH indicator.

According to its useful pH range, for the pH b! A or methyl yellow, methyl orange, methyl red, bromophenol blue, tetrabromophenol blue or bromocresol green can be used. For the medium pH range, Cresol red, Nitricin, bromothymol blue, neutral red, rosolic acid, red OI - naphtholphthalein or phenol red can be used.

PH test; Example Red Cresol dissolved in water and impregnated in 3M paper (angel blotter). After the material was completely impregnated, the matrix was dried.

Preparation of the alkaline phosphatase test zone The activity of alkaline phosphatase in the liquid ! Amniotic is much higher than in normal urine. Therefore, this activity can be used to differentiate and identify the amniotic fluid of the urine in the vaginal secretion using a dry test where direct contact with said secretions results in the formation of different and distinctive colors for the secretions. When the drop! from I the vaginal secretion comes in contact with the reactive area of the matrix, a distinctive color change will identify the content of the secretion and indicate whether it contains amniotic fluid or urine. j Alkaline phosphatase test: Example! The reaction compound of the reagent matrix j for the detection of alkaline phosphatase contains a dry buffered solution of alkaline phosphathase substrate such as p-nitrophenyl phosphatase, indoxyl phosphatase, 4-methylumbelliferyl phosphate and alpha-naphthyl phosphatase and may also contain sensitive indicators such as bromocresol vejrde.

Reference is made to Figures 2-5 illustrating? a diagnostic device to attach it to the pantale'tas of a woman according to the preferred embodiments of the present invention. All of the diagnostic devices shown in Figures 2 to 5 consist of a base layer 22 that includes layers similar to the layers shown in Figure 1 that allow the device to be worn on women's panties, adhering directly to the panties of woman or adhering it to the pantiprotector while the layer with the test area, which will be explained below, is placed directly below the vaginal canal of the woman. In order to see the device, the protective layer is removed from the drawings. ' Figure 2 illustrates a diagnostic device 20 having test zones that are divided into five reagent carrier matrices 24-32 where each reagent carrier matrix has a diagnostic indicator that is capable of identifying a specific substance as it was prepared. below - eg, the creatinine test, protein test, alkaline phosphatase test, etc. ! Figure 3 illustrates a diagnostic device í 40 that has three different 12 test zones. Each zone | It is divided into five different reactive carrier matrices! where each one has a different indicator capable of distinguishing between urine and amniotic fluid. ! Figure 4 illustrates a diagnostic device 50 similar to device 20 where the indicators i 39 different are in 4 reagent carrier matrices 54 -j 58.

Figure 5 illustrates a diagnostic device 60 having test areas 12, each with reagent carrying matrices 62-68 having different indicators to distinguish between urine and amniotic fluid. j As mentioned in the present before, I according to the invention, the device can conterier two or more reactive carrier matrices in lines, circles or any other formation so that at least two indicators change color to be able to distinguish clearly between urine and amniotic fluid. j Reference is now made to Figures 6 a-d illustrating diagnostic pads according to preferred embodiments of the present invention. As mentioned earlier herein, a cushion can be used j so that it is pressed against a moist regular pantyhose or against wet panties, for example, from I way that contact between the skin and the cushion is avoided. Basically, the cushion is made similar to the adhesive cdj, however, there is no adhesive layer on the cushion.

Figure 6a illustrates a cushion 70 having a base layer 72 where different reagent carrier matrices 74-79 are provided with different indicators. í Each reactive carrier matrix is able to identify I certain substance as explained before in the present.

Figure 6b illustrates a cojon with a base layer j 82 in which the reagent carrier dies 94-98 or 104-106 are arranged in different ways. Figures 6c and 6d illustrate further embodiments of cushions 90 and 100 that have fewer areas of reagent carrying matrices, respectively. j Reference is now made to Figure 7 which illustrates a veterinary diagnostic cushion according to another preferred embodiment of the present invention. The veterinary device 120 is very similar to the diagnostic device used in humans, however, the figure shows a top view of the cushion 120 where the adhesive zone ! 122 is located around test areas 124. |

Claims

i CLAIMS!

1. A dry diagnostic device to distinguish between amniotic fluid and urine in 1 female discharge, where the diagnostic device consists of: a base layer; two inert carrier matrices placed in said base layer; dry reagents in said inert carrier matrices wherein said dry reagents are capable of having a chemical reaction with substances in female secretion selected from two or more of a group consisting of creatinine, alkaline phosphatase and total protein, so as to be visually distinguished between amniotic fluid and urine, with the proviso that said reagents are not enzymes or antibodies.

2. The dry diagnostic device like it? disclosed in Claim 1, wherein said carrier matrices consist independently of absorbent material selected from a group of materials consisting of papers containing fibers, woven and non-woven fabrics, modified synthetic and natural polymers, sponge materials, cellulose, fiber glass, micropore membranes, wood, microporous polymer materials consisting of styrene-based copolymers, latex-based, cellulose-based or cotton-based matrices.

3. The dry diagnostic device like! was disclosed in Claim 1, wherein the dry reagents they are able to react with substances present in the amniotic fluid or in the urine, where the substances independently have a greater concentration in the amniotic fluid than in the urine or have a greater concentration in the urine than in the amniotic fluid.

4. The dry diagnostic device as disclosed in Claim 1, wherein the carrier matrix where the reagent is capable of reacting with creatinine is provided consists of two reactive layers; one of which contains fixed creatinine-sensitive dye, with a dye fixing agent and a second layer containing a buffer capable of maintaining one of the mentioned test zones at a relatively high pH value.

5. The dry diagnostic device as disclosed in Claim 3, wherein one of the mentioned carrier matrices consists of a layer containing a creatinine-sensitive dye, a buffer and a dye-binding agent.

6. The dry diagnostic device as disclosed in Claim 4, wherein said creatinine-sensitive dye is selected from a group of dinitro derivatives consisting of 3 '5'-dinitrobenzamide acid, 2'4'-dinitrobenzoic acid, '5' - dinitrobenzotrifluoride, 3 '5'-dinitrobenzamide, 3' 5'-dinitrobenzoyl phenyl glycine, 3,5-dinitrohydroxyphenylpropionic acid.

7. The dry diagnostic device such as; was disclosed in Claim 5, wherein said creatinine-sensitive dye is selected from a group of dinitro derivatives consisting of 3 '5'-dinitrobenzamide acid, 2,4'-dinitrobenzoic acid, 3 '5' - dinitrobenzamide, 3 '5' 3, 5-dinitrohydroxyphenylpropionic acid. |

8. The dry diagnostic device as j was disclosed in Claim 4, wherein said dye fixing agent is a polymerized quaternary ammonium cation (quarts) selected from a group consisting of: polydiallyldimethylammonium chloride, I polymonoalyltrimethylammonium chloride polytrimethylaminoethyl methacrylate chloride, polyvinylbenzyltrimethylammonium chloride, polyvinylmylpyridine chloride.

9. The dry diagnostic device as disclosed in Claim 5, wherein said fixing agent The dye is a polymerized quaternary ammonium cation (quarts) selected from a group consisting of: polydiallyldimethylammonium chloride,! polyimonoalyltrimethylammonium chloride, polytrimethylaminoethyl methacrylate chloride, polyvinylbenzyltrimethylammonium chloride, chloride | of polyvinyl tilpyridine. i

10. The dry diagnostic device as j was disclosed in Claim 4, wherein said creatinine sensitive dye and said dye binding agent were buffered so as to maintain a stable pH in a range of between 9 and 13.5.

11. The dry diagnostic device as disclosed in Claim 5, wherein said creatinine sensitive dye and said dye binding agent are buffered so as to maintain a stable pH in a range of between 9 and 13.5.

12. The dry diagnostic device like! it was disclosed in claim 4, wherein the buffer is selected from a group consisting of: sodium metasilicate, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydroxide - potassium chloride, potassium carbonate, glycine - sodium hydroxide, and sodium borate. j

13. The dry diagnostic device such as; it was disclosed in Claim 5, wherein the shock absorber! it is selected from a group consisting of: sodium metasilicate, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydroxide - potassium chloride, potassium carbonate, glycine - sodium hydroxide, and sodium borate.

14. The dry diagnostic device as disclosed in Claim 1, wherein one of the dry reagents mentioned in one of the carrier matrices is capable j to react with the total protein.

15. The dry diagnostic device as j was disclosed in Claim 14, wherein one of the dry reagents mentioned is a dye selected from a group of 3 ', 3", 5', 5" - tetrabromophenolsulfonphthalein, and bright blue Coomassie, Fast Green, Light Green, j Pyrogallolsulfonphthalein (red pyrogallol), pyrocatecholsulfonphthalein (violet pyrocatechol), 3 ', 3"j dibromo-5', 5" - dichlorophenolsulfonphthalein, fuchsin acid 1, 2,4-dinitro-1-naphthol (yellow martijus) phloxine B, red congo, ethyl orange and methyl orange.

16. The dry diagnostic device as disclosed in Claim 15, which further comprises | a i buffer - selected from a group consisting of potassium citrate, potassium chloride, potassium sulfate, potassium iodine or potassium phosphate.

17. The dry diagnostic device as disclosed in Claim 1, wherein one of the dry reacts in one of the carrier matrices is capable of reacting with the alkaline phosphatase. ^ i

18. The dry diagnostic device as | HE I disclosed in Claim 17, wherein said dry reagent is selected from p-nitrophenyl phosphate, indoxyl phosphate, 4-methylumbelliferyl phosphate and alpha-naphthyl phosphate. !

19. The dry diagnostic device as disclosed in Claim 1, wherein at least two of said carrier matrices are covered by a transparent protective layer.

20. The dry diagnostic device as disclosed in claim 1, wherein a sticky reinforcing layer is provided below said base layer.

21. The dry diagnostic device as disclosed in Claim 20, wherein the sticky: reinforcing layer is adjacent to the outer protective layer.

22. The dry diagnostic device as disclosed in Claim 15, further comprising a metal ion selected from a group consisting of: Copper, Lead, Zinc, Silver.

23. A kit consisting of: the device of claim 1 and a color index configured to allow the distinction between amniotic fluid and urine by observing the colors of the device after coming in contact with the liquid amniotic or urine with the colors of the index.