JP2011501810A - Diagnostic device for identifying membrane rupture during pregnancy - Google Patents

Abstract

A diagnostic device is provided for distinguishing amniotic fluid from urine in a woman's secretions. The device can be used as a panty shield or attached to a panty shield or used on a single pad that is pressed against a substrate with female secretions.
[Selection] Figure 1

Description

The present invention relates to a diagnostic method and a diagnostic apparatus. More particularly, the invention relates to diagnostic tests and devices for identifying membrane rupture during pregnancy.

Delivery varies from woman to woman, and it is difficult to specify when to begin delivery. The whole process of delivery is a combination of several physiological changes and events that eventually occur in the body to deliver the child, rather than a single phenomenon. One of those changes or events is a rupture of the membrane surrounding the child in the uterus, usually an obvious sign that labor is imminent. The rupture of the membrane suddenly occurs, and obviously a large amount of amniotic fluid flows. Often, however, a pregnant woman may have a ruptured membrane and may or may not be able to determine if the membrane has truly ruptured.

There are two main reasons for this. First, because the fetal head can act like a cork at the opening of the uterus, the amniotic fluid is released slowly without spewing out. In addition, amniotic fluid leaks from a small opening in the uterus, and the pregnant woman does not feel the initial contraction until several hours later. Second, it is often difficult for women to control urination in late pregnancy. For this reason, if women are accustomed to urinating small amounts of urine, amniotic fluid from the vagina flows slowly and often does not notice if it leaks unintentionally.

Misdiagnosis and missed diagnosis of leakage of amniotic fluid, especially in high-risk pregnancies, increases the risk to pregnant women and the fetus without proper treatment. The risk to newborns of unknowingly leaking amniotic fluid includes fetal asphyxia, infection and childbirth before the scheduled date, which can lead to very dangerous consequences for the mother and the fetus.

Currently, the only available method for pregnant women to detect amniotic fluid at home is accomplished using an expensive panty liner self-test diagnostic kit. Thus, almost all pregnant women choose to see a doctor in a hospital or emergency room. In hospitals, the tests that are currently available to identify amniotic fluid are inconvenient for patients because they are invasive, obscure or expensive. By using an accurate, fast and inexpensive device that also functions as a tool used by a physician to perform a clinic / hospital examination, the possibility of rupturing the membrane without being noticed at home needs to be greatly reduced.

Among several conventional methods currently available to physicians, a few examples are described herein. The most extensive method for analyzing urine composition is to use a technique known as “immersion and reading assay strip”, “urine test strip”. A urine test strip is an assay strip consisting of reagents bound to a reagent carrier matrix (pad) on a strip. Usually, the piece itself is formed from a polymeric material (eg, polyethylene, polycarbonate or polystyrene). Each carrier matrix has a different reagent therein. The urine test paper is taken out by being immersed in a urine sample. When the reagent embedded in the matrix comes into contact with the urine sample, a color reaction occurs. Urine strips can be designed as a single pad test strip (for assaying one analyte) or as a composite pad test strip (for assaying several different samples simultaneously). Urine strips can be used manually or with a suitable chemical analyzer. Various shapes of reagent pieces for performing many analyzes of specimens simultaneously or sequentially are disclosed in US Pat. No. 4,595,439, US Pat. No. 4,526,753, US Pat. No. 4,160. U.S. Pat. No. 3,123,443, U.S. Pat. No. 3,212,855, U.S. Pat. No. 3,814,668, U.S. Pat. No. 4,038,485, U.S. Pat. This is disclosed in Japanese Patent No. 3,531,254.

Some of such test devices are commercially available. Trademarks, CLINISTIX, MULTISTIX, KEOSTIX, N-MULTISTIX, DISTIX, DEXTROSTIX, AUTION STICKS, and CHEMSTRIP are part of the urine test strip.

The reagent carrier matrix is typically an absorbent material in which a liquid sample can move through the matrix. This movement of the liquid sample is dependent on the capillary force that is formed in the matrix. As the liquid moves through the matrix, the liquid contacts the chemical reagent composition impregnated within the matrix. Thereafter, a detectable and measurable color transition occurs. If the urine paper can measure several analytes simultaneously, the color change within each reagent carrier matrix can be correlated with the amount of various analytes in the liquid sample. Manual analysis of the results requires comparing the color transition of the examination on the urine paper with a color chart.

The reagent carrier matrix material can be any material that incorporates the chemical reagents necessary to perform the assay of interest. Preferred matrices must be inert to the reagents and must not alter the sample or test results. The reagent carrier matrix can consist of many materials, some of which include fiber-containing papers such as filter paper, woven and non-woven fabrics, synthetic or modified natural polymers, sponge materials, cellulose, glass fibers, Perforated membrane, wood. Reagent matrices can differ in terms of roughness and smoothness as well as flexibility and strength. The use of various matrices is described in US Pat. No. 3,846,247, US Pat. No. 3,552,928, US Pat. No. 3,802,842, US Pat. No. 3,418,083. Are listed.

Sewell DL et al., In particular, describes the cost of using a urine test strip as a screening method for urinalysis in a scientific paper published in the American Journal of Clinical Pathology 83 (6) 740-743, 1985. . The author states that the method using urine test strips "costs about $ 0.76 for the reagent". Various devices are described in the literature regarding the measurement of specific urine specimens using a reagent carrier matrix (filter paper, microcapsules, urine test paper, etc.). Prior art assay devices include dry tablets, urine test strips or other techniques for analytical urine components. U.S. Pat. No. 4,147,514 describes the detection of ketone bodies and U.S. Pat. No. 3,146,070 describes a carrier (urine test strip) impregnated with a pH indicator for pH measurement. A dry form chemical composition is disclosed. Methods, compositions, and test devices for measuring the ionic strength or specific gravity of a sample for measurement (eg, urine) are disclosed in US Pat. Nos. 4,318,709 and 5,403,744.

The Jaffe method is a well-known method for measuring creatinine. The method includes an aqueous alkaline picrate solution in the orange-red form. US Pat. No. 6,001,656 discloses a device for assaying creatinine in a fluid test sample. Improvements to this patent include the inclusion of one or more selected reagent dosage forms of quinoline. Another method for the determination of creatinine is described by Benedict and Behre in Journal of Biological Chemistry (1936) and involves the reaction of creatinine with 3,5-dinitrobenzoic acid in an alkaline medium. Other methods, compositions and test apparatus for determining creatinine in a liquid sample such as urine are disclosed in US Pat. Nos. 4,215,197 (using enzyme composition) and 5,662,867. No. 5,733,787. Suitable materials for the detection of creatinine include picric acid, 3,5-dinitrobenzoic acid, 3,4-dinitrobenzoic acid, 2,4-dinitrobenzene sulfonic acid, (3,5-dinitrobenzyl) alcohol, (3,5-dinitrobenzo) -nitrile, (3,5-dinitrobenz) amide and N, N-diethyl- (3,5-dinitrobenz) amide.

Various methods for measuring fluid proteins have been reported. These methods include the Burette method, the Raleigh method, the Kjeldahl method, the dye combination method, the fluorometric method and the UV method. Of these methods, J. Org. Lab. Clin. Med. , 11, 981 (1926), the Kingbury-Clark method, and Clin. Chim. Actu, 5, 757 (1960), the Meulemans method and Anal. Biochem. 72, 248 (1967), the Coomassie Brilliant Blue method is widely used. The Bradford dye assay for protein quantification (US Pat. No. 4,023,933) is commonly used in almost all biochemical laboratories. Usually proteins are substances, mainly pigments such as Coomassie brilliant blue, bromophenol blue (tetrabromophenol blue), and eosin, and metal ions such as copper (II), lead (II), zinc (II) and Interacts with silver (I). Addition of a protein-containing aqueous solution for the reaction between the dye and the metal ion gives a spectral change to the aqueous dye-metal ion solution. More protein indicators include the merocyanine and nitro or nitroso substituted polyhalogenated phenol sulfonephthaleins disclosed in US Pat. No. 5,279,790 and those described above. Other protein indicators include Fast Green FCF, Light Green SF, pyrogallol red and pyrocatechol violet, bromochlorophenol blue (3 ', 3 "-dibromo-5', 5" -dichlorophenolsulfonphthalein), basic fuchsin , Basic violet, maltose yellow, Phloxine B, methyl yellow, Congo red, methyl orange and ethyl orange (4- (4-dimethylaminophenylazo) benzenesulfonic acid). US Pat. No. 5,424,215, US Pat. No. 5,593,895, US Pat. No. 6,815,210, US Pat. No. 4,960,710, US Pat. No. 3,485 No. 5,877, US Pat. No. 5,087,575, and US Pat. No. 4,023,933 relate to protein measurement of aqueous solutions such as urine using a reagent system that can be used in the urine test paper method.

  Various urine test strips used for urinalysis include tests for urobilinogen. ROCH diagnostic CHEMSTRIP and Bayer diagnostic MULTISTIX are typical examples of such products that include testing for urobilinogen. The classic urobilinogen test developed by Paul Ehrlich in 1901 uses paradimethylaminobenzaldehyde which produces a brown-orange-red color with Ehrlich reagent in a strongly acidic medium. As further background for urobilinogen, the Ehrlich reaction and urobilinogen assay are described in Tietz W.H. B. It is described in the text of clinical chemistry of Saunders Company. Examples of urine test papers for the measurement of urobilinogen based on Ehrlich reaction and diazonium conjugation reaction include US Pat. No. 3,853,466, US Pat. No. 3,630,680, US Pat. No. 4,665. No. 038, U.S. Pat. No. 4,290,771, U.S. Pat. No. 3,989,462, U.S. Pat. No. 3,814,586. Further, US Pat. Nos. 4,158,546 and 3,447,905 relate to the measurement of urobilinogen in aqueous solutions, such as urine, using a reagent system useful in the urine test paper method.

The earliest method for testing alkaline phosphatase was introduced by Kay in 1930. Later, a popular assay method for the measurement of alkaline phosphatase using p-nitrophenyl phosphate was introduced in 1946 by Bessey, Lowry and Block. The fact is that after exposure to a fluid containing alkaline phosphatase, the colorless p-nitrophenyl phosphate is catalytically hydrolyzed to the yellow colored product p-nitrophenol (and phosphate). Depends on. Thus, the enzyme concentration is measured by increasing the yellow intensity of the reaction product. Alkaline phosphatase activity is naturally present in unpasteurized milk and the enzyme is denatured after sterilization. As such, alkaline phosphatase activity is used as an indicator for proper sterilization of milk. Such a dry test of alkaline phosphatase activity in milk is available from MACHEREY-NAGEL GMBH & CO. It is made by PHOSPHATESMO ML.

The presence of hemoglobin in the urine is called hemoglobinuria, and such a condition can occur as a result of lysis of red blood cells (RBS) in the urinary tract. The term hematuria is used when complete RBS is present in the urine. This condition can cause kidney or urogenital bleeding. The most widely used test for the detection of blood in urine or stool relies on the fact that heme protein can act as a peroxidase. This reaction requires a hydrogen donor molecule. Typical examples of products that include tests for blood detection are MULTISTIX 10SG, HEMOCULT II, and AUTION STICKS.

Recently, US Pat. No. 4,357,945 issued on Nov. 9, 1982, “A device for examining and rupturing amniotic membrane” (Janco) describes a finger-engaging device with a pH indicator. To do. When exposed to fluid, the indicator changes color when the amniotic membrane ruptures. US Pat. No. 5,425,377, issued June 20, 1995, “Measurement of body fluid pH” (Caillouette) describes a swab structure on a stick with a pH indicator for the measurement of vaginal fluid pH. .

Several other methods are provided as follows: US Pat. No. 5,554,504, issued September 10, 1996, “Diagnostic Methods for Detecting Female Membrane Rupture” (Rutanen) ) Describes the detection of insulin-like growth factor binding protein 1 in vaginal secretion samples. US Pat. No. 5,281,522, issued Jan. 25, 1994, “Reagents and kits for determination of fetal fibronectin in vaginal samples” (Seniei et al.) A kit is described for detection of membrane rupture by exposing it to an antibody such as a fibronectin antibody and an anti-fibronectin antibody. U.S. Pat. No. 5,096,830 issued March 17, 1992, “Birth and membrane rupture test before scheduled date” (Seniei et al.) Removes the sample from the vaginal cavity and insoluble anti-fetal antigen antibody is attached. A method is described in which fetal membrane rupture is measured by contacting it with a support and fetal antibodies bound to the support are measured.

  Several devices are known, including panty shields or tampons with pH indicators. Some examples of patents are listed below: US Pat. No. 6,149,590, issued Nov. 21, 2000, “System for identifying premature rupture of membranes during pregnancy” (Smith et al.) , A pad having an upper outer layer, a lower outer layer and an intermediate pH reaction component is described. US Pat. No. 6,921,647 and US Pat. No. 6,627,394 “Secret Monitoring Products and Diagnostic Pads” (Kritzman et al.) Describe a pad with a pH sensitive indicator for detecting leakage of amniotic fluid. Describe. US Pat. No. 5,217,444 “Absorptive Tampon” (Schoenfeld) discloses an absorbent material comprising a pH indicator material that indicates the acidity or alkalinity of the liquid in contact by a change in color.

  It should be noted that a test that measures only the pH of the vaginal secretion to distinguish urine from amniotic fluid is inaccurate for the following reasons. 1) pH of urine can vary from 4.5 to 8 depending on renal homeostasis activity and water intake, 2) pH of amniotic fluid varies from 6.9 to 7.15 in late pregnancy To do. This overlapping range of pH can lead to false diagnoses that can cause medical complications.

Immunochromatographic tests based on antibodies that detect specific proteins in amniotic fluid or urine are not readily available to the general public because they are expensive due to the monoclonal / polyclonal antibodies they contain. Usually, enzymatic methods are also expensive because of the high production costs of the enzymes.

  The object of the present invention is a unique apparatus for distinguishing amniotic fluid from urine based on the analysis of one or more specimens using dry chemical reactions without the need for enzymatic or immunological methods. And providing a method.

Another object of the present invention is to provide a unique device and method for distinguishing between urine and amniotic fluid in a simple, inexpensive and short time, which ultimately helps to resolve the dilemma of whether delivery begins immediately. There is to do. A woman or medical caregiver can observe whether the first drop of leaked liquid contains amniotic fluid or urine immediately after contact with the device.

  Yet another object of the present invention is to provide a unique device and method for distinguishing between urine and amniotic fluid that can be used on human or non-human pregnant female subjects. Since animals cannot account for the situation, it can be unpredictable to measure the onset of labor in non-human women.

  Yet another object of the invention is the implementation of care for rapid chemical analysis of body fluids such as urine, saliva, sweat, cerebrospinal fluid (CSF), milk or fluids from other sources.

Thus, according to a preferred embodiment of the present invention,
A dry diagnostic device for distinguishing amniotic fluid from urine in a woman's secretions,
The base layer,
At least two inert support matrices provided in the base layer;
A dry reagent provided on the at least two inert support matrices,
The dry reagent visually distinguishes the amniotic fluid from the urine by causing a chemical reaction with a plurality of substances in the woman's secretions,
A dry diagnostic apparatus is provided in which the dry reagent in each of the at least two support matrices can react with different substances in the plurality of substances.

In yet another preferred embodiment of the invention, the at least two support matrices are: fiber-containing paper, woven and non-woven, synthetic or modified natural polymer, sponge material, cellulose, glass fiber, microporous membrane, wood , Made from an absorbent material selected from the group of materials such as microporous polymer materials (such as styrene-based copolymers, latex-based, cellulose-based or cotton-based matrices).

In still another preferred embodiment of the present invention, the dry reagent is capable of reacting with a substance present in the amniotic fluid or the urine, and the substance is more than the original concentration in the amniotic fluid or the urine. Has a significantly higher concentration.

In yet another preferred embodiment of the invention, said substance comprises substances such as creatinine, alkaline phosphatase, total protein, urea, urobilinogen and blood.

In yet another preferred embodiment of the invention, one of the dry reagents in one of the at least two support matrices is capable of reacting with creatinine.

In still another preferred embodiment of the present invention, one of the support matrices includes two reagent layers, one of the reagent layers includes a creatinine sensitive dye fixed to a dye fixative, The reagent layer includes a buffer that allows one of the test areas to be kept at a relatively high pH value.

In yet another preferred embodiment of the present invention, one of the support matrices comprises a single layer comprising a creatinine sensitive dye, a buffer and a dye fixative.

In yet another preferred embodiment of the invention, the creatinine sensitive dye is 3′5′-dinitrobenzoic acid, 2′4′-dinitrobenzoic acid, 3′5′-dinitrobenzotrifluoride, 3′5. Selected from the group of dinitro derivatives such as' -dinitrobenzamide, 3'5'-dinitrobenzoylphenylglycine, 3,5-dinitrohydroxyphenylpropionic acid.

In still another preferred embodiment of the present invention, the dye fixing agent is polydiallyldimethylammonium chloride, polymonoallyltrimethylammonium chloride, polytrimethylaminoethyl methacrylate chloride, polyvinylbenzyltrimethylammonium chloride, polyvinylmethylpyridine chloride. Selected from polymerized quaternary ammonium cations (quats).

In yet another preferred embodiment of the invention, the creatinine sensitive dye and the dye fixative are buffered to maintain a stable pH within the range of about 9 to 13.5.

In still another preferred embodiment of the present invention, the buffering agent is sodium metasilicate, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydroxide-potassium chloride, potassium carbonate, glycine-sodium hydroxide, And selected from the group of sodium borate.

In yet another preferred embodiment of the invention, one of the dry reagents in one of the support matrices is capable of reacting with total protein.

In yet another preferred embodiment of the present invention, one of the dry reagents is 3 ′, 3 ″, 5 ′, 5 ″ -tetrabromophenolsulfonephthalein, Coomassie Brilliant Blue, Fast Green, Light Green Pyrogallol sulfonephthalein (pyrogallol red), pyrocatechol sulfophthalein (pyrocatechol violet), 3 ', 3 "-dibromo-5', 5" -dichlorophenol sulfonephthalein, fucic acid, 2,4-dinitro- 1-Naphthol (Martinus Yellow) Phloxine B, Congo Red, ethyl orange and methyl orange are selected from the group.

Yet another preferred embodiment of the present invention further comprises a buffering agent such as potassium citrate, potassium chloride, potassium sulfate, potassium iodate or potassium phosphate.

Yet another preferred embodiment of the present invention further comprises a metal ion selected from copper, lead, zinc, silver.

In yet another preferred embodiment of the present invention, one of the dry reagents in one of the support matrices is capable of reacting with alkaline phosphatase.

In yet another preferred embodiment of the invention, one of the dry reagents is p-nitrophenyl phosphate, indoxyl phosphate, 4-methylumbelliferyl phosphate and alpha-naphthyl-phosphate. A substrate for an alkali phosphate selected from salts.

In yet another preferred embodiment of the invention, one of the dry reagents is a pH sensitive reagent.

In yet another preferred embodiment of the invention, the dry reagent does not exit the support matrix.

In yet another preferred embodiment of the invention, the at least two support matrices are covered with a protective layer.

In yet another preferred embodiment of the invention, the protective layer is transparent.

In yet another preferred embodiment of the invention, the protective layer is made of a thin “one-way structure” membrane that is permeable to liquid flowing into the at least two support matrices, and the reagent is externally removed from the device. Prevent it from flowing.

In still another preferred embodiment of the present invention, an adhesive backing layer is provided below the base layer.

In yet another preferred embodiment of the invention, the adhesive backing layer is provided proximate to the outer protective layer.

In yet another preferred embodiment of the invention, the at least two supported reagent matrices are arranged in substantially parallel rows.

In still another preferred embodiment of the present invention, the at least two reagent-carrying matrices are arranged concentrically.

In still another preferred embodiment of the present invention, the diagnosis is performed when at least two of the examination regions are opposed to the vaginal canal of a female animal by surrounding the at least two reagent-carrying matrices with an adhesive substance. Allows the device to adhere.

Several embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings. In making detailed reference to these drawings, it should be emphasized that they exist to provide the most useful and understandable description of the principles and conceptual aspects of the present invention. In so doing, there is no aim to show structural details of the invention beyond what is necessary for a basic understanding of the invention, and the description and drawings actually illustrate how some embodiments of the invention may be practiced. It is for clarifying what to do.

1 shows a cross-sectional view of a diagnostic device according to a preferred embodiment of the present invention. 1 shows a diagnostic device for attaching to female panties according to a preferred embodiment of the present invention. 1 shows a diagnostic device for attaching to female panties according to a preferred embodiment of the present invention. 1 shows a diagnostic device for attaching to female panties according to a preferred embodiment of the present invention. 1 shows a diagnostic device for attaching to female panties according to a preferred embodiment of the present invention. (A)-(D) show the diagnostic pad by suitable embodiment of this invention. Figure 5 shows a veterinary diagnostic pad according to another preferred embodiment of the present invention.

  Before describing at least one embodiment of the present invention in detail, it is to be understood that the present invention is not necessarily limited to the application described in the following description or illustrated examples. The invention may be practiced or carried out in other embodiments or in various ways.

  The terms “including” (comprises, comprising, includes, including), and “having” and their contiguous terms mean “including but not limited to”.

  The term “consisting of” has the same meaning as “including and limited to”.

  The term “consisting essentially of” means that a composition, method, or method, unless further materials, steps, and / or parts significantly change the fundamental and novel characteristics of the claimed composition, method, or structure. Or it means that the structure may contain further materials, steps and / or parts.

  As used herein, the singular forms “a” (a, an) and “the” include the plural unless the context clearly dictates otherwise. For example, the term “compound” or “at least one compound” can also include a plurality of compounds and mixtures thereof.

Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be taken into account in particular as disclosing all possible sub-ranges as well as individual numerical values within that range.

It should be understood that certain features of the invention described in the text of another embodiment may be provided in combination in a single embodiment for increased clarity. On the contrary, for the sake of brevity, the various features of the invention described in the context of a single embodiment are preferred separately or in any suitable sub-combination or other described embodiment. Can be provided. Certain features that are described in the context of various embodiments are not considered basic features of the embodiments unless the embodiments operate without these elements.

The present invention provides a unique and novel device for easily and quickly distinguishing between urine and amniotic fluid so that pregnant women can know the status of pregnancy. In accordance with one aspect of the present invention, a device similar to a “stick bandage”, which is a treatable strip containing a sensitive chemical indicator, is used in the last few months of pregnancy. The strip is of relatively small size and consists of at least two layers (absorbent material including an adhesive backing and an indicator). A woman using the device simply attaches the device to a panty that has an examination area facing the body. When a woman uses a panty liner, a strip can be placed thereon. If the amniotic fluid leaks due to rapid chemical reaction in the examination area of the device, a distinct color change will occur, and a distinctly different color change will occur when urine contacts the examination area. The device of the present invention acts as a diagnostic tool that allows the customer to detect whether the secretions contain only amniotic fluid or urine, by means of the well-known color index supplied to the device.

According to a second aspect of the invention, a pad-like device containing a sensitive indicator is used independently of contact with the female body. The device can be used by pressing against wet underwear or a wet panty liner after noting fluid leakage. This causes a chemical reaction in the reagent-carrying matrix and a color change almost immediately when the reagent in the reagent-carrying matrix contacts the body fluid and the leaked liquid contacts the diagnostic device. Also, a woman (or caregiver) using this device can see if the liquid contains only amniotic fluid or urine by a well-known color index supplied to the device.

According to a third aspect of the present invention, a device similar to a panty liner includes a sensitivity indicator that allows discrimination between amniotic fluid and urine. Pregnant women spend their daily life wearing this product on their underwear. The indicator reagent area must be placed directly opposite the vagina. Thus, the sensitive panty liner is in intimate contact with female body fluids.

In all aspects of the invention, the sample can be transferred to a suitable container for storage when storage is required for the next collection. Alternatively, immediate processing of the sample can be performed. When used, the sample is placed directly on the reagent-bearing matrix of the device and the test is performed in a small collection sample.

  In another aspect of the invention, a veterinary device is provided. Devices containing sensitive reagents are attached directly to the opening of the vagina of non-human women. This can be easily achieved by a bandage design where the sticky area is around or on the side of the reagent-bearing matrix test area. In this way, intimate contact with animal body fluids is achieved. Using a device that is not in direct contact with the animal's body can be accomplished by obtaining a fluid sample or a suction or cleaning device attached to a swab having a fibrous tip and applying it to an indicator area on the device.

According to a preferred embodiment of the present invention, the determination of whether amniotic fluid or urine is present in a woman's secretion is a number of non-enzymatic and non-immunological measures that measure either urine or amniotic fluid by a prominent color. Depending on the reaction isolated. Currently, in most of the available tests for confirmation of the cause of wetness during pregnancy, the concentration difference of only one specimen of urine or amniotic fluid is measured. In the present invention, the presence of at least two of the substances protein, creatinine, urea, urobilinogen, blood and alkaline phosphatase is identified as well as pH. Basically, while creatinine, urea and urobilinogen have high urine concentrations in amniotic fluid, it has been identified that proteins and alkaline phosphatase have high amniotic fluid concentrations associated with urine. Blood can be present in amniotic fluid at a higher concentration than normal urine. A chemical that can be used to distinguish fluids according to a preferred embodiment of the present invention binds or reacts with one of the materials shown here, or a particular environment acts through a change in spectroscopic properties. To do.

Therefore, the diagnostic apparatus including the reagent-carrying matrix provided with the chemical indicator according to the present invention can detect at least two substances among proteins, creatinine, urobilinogen, urea, blood, and alkaline phosphatase as well as the pH value. The method of the present invention is based on the following facts. 1) The total protein concentration of amniotic fluid is usually about 15 times higher than that of urine; 2) The concentrations of creatinine and urobilinogen in urine are usually about 10 times higher than those in amniotic fluid; 3) urine The concentration of urea is usually more than twice as high in amniotic fluid; 4) The concentration of alkaline phosphatase in amniotic fluid is usually more than about seven times higher than the concentration of urine; 5) emerges during fetal membrane rupture Amniotic fluid can contain blood, as opposed to urine in healthy women. Thus, even if a small amount of amniotic fluid is present in the vaginal secretions and contacts the pad of the present invention, the pad allows diagnosis of fetal membrane rupture with very high accuracy. Other substances with similar shading can also be detected.

The invention is further illustrated by the following example of a device with a sensitive indicator for detection and fractionation of urine or amniotic fluid leakage.

Reference is made to FIG. 1, which shows a diagnostic device according to a preferred embodiment of the present invention in a top view and a cross-sectional view, respectively. The device 10 has at least two and preferably several reagent carrying matrices 12. FIG. 1 shows a device covered with a first layer 14 of a soft and comfortable material that is suitable for being attached to a woman's body and can therefore be worn on a woman's panties and does not irritate the skin when contacted. Indicates. The first layer 14 prevents direct contact between the reagent 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 differentiation of the colors formed in the layers below it.

The device 10 further includes a support substrate 16 that can support a reagent-carrying matrix 12 capable of performing the desired assay. The reagent carrying matrix 12 is preferably an absorbent material that allows the liquid sample to move through the matrix. The reagent carrying matrix 12 must be inert with respect to the reagents and must not change the sample or test results.

Optionally, the reagent-carrying matrix 12 can be composed of many materials such as fiber-containing papers such as filter paper, woven and non-woven fabrics, synthetic or modified natural polymers, sponge materials, cellulose, glass fibers, microporous membranes, wood and the like. The further material can be a microporous polymer material such as a styrene-based copolymer, latex-based, cellulose-based or cotton-based matrix.

The reagent-bearing matrix can also vary in properties such as roughness, smoothness, flexibility and hardness.

It should be noted that in the manufacturing process, the reagent-carrying matrix can consist of multiple layers, some of which carry different reagents in different areas of the examination area. Any combination of a supporting base layer and a further matrix carrying an indicator is included within the scope of the present invention, and the scope of the present invention is in no way limited.

Below the support base layer 16, an adhesive backing layer 18 is provided and an adjacent outer protective layer 20 is also provided. These two layers are essentially similar to the layers provided on the protective panty shield.

The reagent carrying matrix 12 is preferably composed of groups, each group comprising an indicator capable of indicating one of the aforementioned substances. The following are methods for preparing chemical reagents and test areas for each.

Preparation of the creatinine test area The creatinine concentration in urine is usually about 10 times higher than the amniotic fluid concentration. The creatinine dryness test can be made in two arbitrary ways:
-A two reagent layer system comprising a fixed creatinine sensitive dye placed on one matrix and a buffer that keeps the system at a relatively high pH value is placed on the second matrix.
-One reagent layer system containing creatinine sensitive dye, buffer, stabilizer all placed in one matrix.

Creatinine sensitive dyes are 3'5'-dinitrobenzoic acid, 2'4'-dinitrobenzoic acid, 3'5'-dinitrobenzotrifluoride, 3'5'-dinitrobenzamide, 3'5'-dinitrobenzoyl-phenyl. Can be one of dinitro derivatives such as glycine, 3,5-dinitrohydroxyphenylpropionic acid.

Sensitive dye fixing agent is preferably polymerized quaternary ammonium such as polydiallyldimethylammonium chloride (polyDADMAC), polymonoallyltrimethylammonium chloride, polytrimethylaminoethyl methacrylate chloride, polyvinylbenzyltrimethylammonium chloride, polyvinylmethylpyridine chloride Contains cations (quat).

The buffer should be a strong base capable of maintaining a stable pH in the range of about 9-13.5, for example, the buffer can be sodium metasilicate, sodium hydroxide, potassium hydroxide, hydroxide Calcium, sodium hydroxide-potassium chloride, potassium carbonate, glycine-sodium hydroxide, and sodium borate.

Optionally, a non-volatile solid reagent is added to the sensitive dye to enhance the reaction.

Two reagent layer systems; Example 1
A creatinine test indicator consisting of two reagent-bearing matrices was placed on the diagnostic device substrate. As a stabilizer dissolved in water, a first matrix (which can be Whatman filter paper or napkin paper) was impregnated with a creatinine sensitive dye, 3′5′-dinitrobenzoic acid (dBA) and polyDADMAC (optional). . The dBA stock solution was mixed with sodium carbonate buffer. The second reagent matrix was impregnated with sodium metasilicate buffer. After both reagent matrices were dried, the first reagent matrix was placed on a second reagent matrix that was impregnated with sodium metasilicate buffer. Both the first and second dry reagent matrices are in close contact with the base layer of the device.

After a drop of vaginal discharge has contacted the dry reagent-carrying matrix, a different color change identifies the fluid and indicates whether it contains amniotic fluid or urine.

Two reagent layer systems; Example 2
The two reagent-absorbing matrices are each impregnated in one solution, dried, and adhered to the substrate of the device. The first reagent matrix consists of a solution 1 consisting of dBA as a creatinine sensitive dye (stock solution dissolved in sodium hydroxide), a non-volatile solid reagent and (optionally) polyDADMAC as a stabilizer. Impregnated. The second reagent matrix was impregnated with solution 2 containing sodium hydroxide. Both reagent matrices were dried. The dry reagent matrix with solution 1 is placed on the dry reagent matrix with solution 2 and both are in close contact with the substrate of the device.

One reagent layer system; Example 1
As described above, a “one reagent layer system” has the same rational as a “two reagent layer system”, but includes all chemical products on one reagent carrying matrix. In either case, the goal is to distinguish urine and amniotic fluid using creatinine concentration.

The creatinine testing device is prepared from the same absorbent material and support carrier in the two reagent layer systems. The reagent-carrying matrix consists of a creatinine sensitive dye, a non-volatile solid reagent, a stabilizer (optional), and a buffer that is dried and mounted on the carrier matrix. Use 3'5'-dinitrobenzoic acid as a sensitive dye (stock solution made in acetonitrile), polyDADMAC as stabilizer (optional), and potassium hydroxide as buffer (in ethyl alcohol solution). Is preferred. The reagents were dried all at once and were ready for urine or amniotic fluid sample testing.

One reagent layer system; Example 2
An emulsion of 3'5'-dinitrobenzoic acid, polyDADMAC (optional), styrene acrylic acid, sodium metasilicate and a non-volatile solid reagent was prepared. A thin layer of the emulsion was spread on a latex-based matrix and dried. The dry matrix was intimately adhered onto the support substrate polymer of the device.

Preparation of total protein test area The concentration of total protein in the amniotic fluid is usually about 15 times or more the concentration in urine. The interaction of proteins with substances, mainly dyes and metal ions, causes spectral changes in dye-metal ion solutions. In the implementation of the apparatus of the present invention, 3 ′, 3 ″, 5 ′, 5 ″ -tetrabromophenol sulfonephthalein, Coomassie brilliant blue, first green, light green, pyrogallol sulfonephthalein (pyrogallol red), pyrocatechol sulfone Phthalein (pyrocatechol violet), 3 ′, 3 ″ -dibromo-5 ′, 5 ″ -dichlorophenolsulfonephthalein, fuchsic acid, 2,4-dinitro-1-naphthol (Malto yellow), copper, lead, Zinc, silver, Phloxine B, Congo red, ethyl orange and methyl orange can be used.

The reagent-bearing matrix of the total protein testing device may be an absorbent carrier, such as a microporous polymeric material (eg, a styrene-based copolymer), latex-based, cellulose-based or cotton-based, which can be one of the matrices described above. Matrix. The reagent-bearing matrix is a polymerized urethane-based compound (described in US Pat. No. 5,124,266) that incorporates an indicator reagent compound that can interact with a protein to produce a visually detectable reaction. ).

To reduce variability due to differences in urine density, the test solution may contain a low pH potassium salt-based buffer such as potassium citrate, potassium chloride, potassium sulfate, potassium iodate or potassium phosphate.

Total protein test; Examples The total protein solution test consists of a combination of two solutions (dye reagent solution and buffer). As the dye solution, 3 ′, 3 ″, 5 ′, 5 ″ tetrabromophenolsulfonephthalein was used and was dissolved in a weak organic acid such as citric acid. The buffer, potassium citrate, is titrated with the same weak organic acid to a low pH value of about 3.5.

The dye solution was then diluted with buffer on a wide scale. After dilution, a very thin layer of the resulting solution was spread on the reagent-carrying matrix and dried. A separate color reaction can distinguish between urine and amniotic fluid while urine and amniotic fluid come into contact with the reagents embedded in the device.

Preparation of pH test region As mentioned above, this preparation is placed on any type of absorbent reagent-bearing matrix. The reagent matrix is impregnated with a pH indicator for measuring fluid pH. While amniotic fluid pH varies from 6.9 to 7.15 in late pregnancy, the pH of urine can vary from 4.5 to 8. This pH range is checked by one indicator or two different pH indicators (low pH indicator and intermediate pH indicator).

As their useful pH range, methyl yellow, methyl orange, methyl red, bromophenol blue, tetrabromophenol blue or bromcresol green are used as low pH. As the intermediate pH range, cresol red, nitridine, bromothymol blue, neutral red, rosoleic acid, α-naphtholphthalein or phenol red can be used.

pH test; Example Cresol Red was dissolved in water and impregnated in 3M paper (absorbent paper ra-reeve angel®). After the material is completely impregnated, the matrix is dried.

Preparation of alkaline phosphatase test area Alkaline phosphatase activity in amniotic fluid is much higher than normal urine. Therefore, this activity distinguishes and distinguishes amniotic fluid from urine in vaginal secretions by using a dry test where the distinct and different color forms for secretions cause direct contact with such secretions. Can be used to When a drop of vaginal secretion touches the dry matrix reaction area, a distinct color change identifies the secretion content and indicates whether it contains amniotic fluid or urine.

Alkaline phosphatase test; Examples Reaction reagent matrix composition for detection of alkaline phosphatase includes p-nitrophenyl phosphate, indoxyl phosphate, 4-methylumbelliferyl phosphate and alpha-naphthyl-phosphorus Contains a dry buffer solution of an alkaline phosphatase substrate such as an acid salt and may contain a sensitive indicator such as bromocresol green.

Reference is made to FIGS. 2-5 illustrating a diagnostic apparatus for depositing on female panties in accordance with a preferred embodiment of the present invention. All diagnostic devices shown in FIGS. 2-5 include a base layer 22, which includes a layer similar to the layer shown in FIG. 1, and is used for female panties by the layer shown in FIG. A device is used in women's panties that is directly attached to the women's panties, or that is attached on the panty shield and in which a layer having the examination area described below is placed directly under the woman's vaginal canal Make it possible. In order to view the device, the protective layer is omitted from the drawing. FIG. 2 shows a diagnostic device 20 having an examination region that is divided into five reagent-bearing matrices 24-32, each reagent-bearing matrix being able to identify a specific substance as described in detail above. Indicators such as creatinine test, protein test, alkaline phosphatase test, etc. are provided.

FIG. 3 shows a diagnostic device 40 having three different examination areas 12. Each region is divided into five different reagent-bearing matrices, each with a different indicator that can distinguish between urine and amniotic fluid.

FIG. 4 shows a diagnostic device 50 that is similar to device 20, with different indicators in four reagent carrying matrices 54-58.

FIG. 5 shows a diagnostic device 60 having an examination region 12 having reagent-bearing matrices 62-68, each with a different indicator, to distinguish urine and amniotic fluid.

As described herein, according to the present invention, the device can include two or more reagent-carrying matrices in a linear, circular or any other form, clearly distinguishing between urine and amniotic fluid. It is possible that at least two indicators change color.

Reference is now made to FIGS. 6a-6d showing a diagnostic pad according to a preferred embodiment of the present invention. As described above, the pad can be used in a manner that is pressed against a normal wet panty shield or wet panties, for example, to prevent contact between the skin and the pad. Basically, the pad is constructed in the same way as an attached one, but this pad has no attached layer.

FIG. 6a shows a pad 70 having a base layer 72 on different reagent carrying matrices 74-79 with different indicators. As described above, each reagent-bearing matrix can identify a specific substance.

FIG. 6b shows a pad having a base layer 82 in which reagent carrying matrices 94-98 or 104-106 are arranged in different arrangements. Figures 6c and 6d show a further embodiment of pads 90, 100, each with fewer reagent carrying matrix regions.

Reference is now made to FIG. 7 showing a veterinary diagnostic pad according to another preferred embodiment of the present invention. The veterinary device 120 is basically similar to the diagnostic device used for humans, but this figure shows a top view of the pad 120 with the adhesive area 122 around the examination area 124.

Claims (27)

A dry diagnostic device for distinguishing amniotic fluid from urine in a woman's secretions,
The base layer,
At least two inert support matrices provided in the base layer;
A dry reagent provided on the at least two inert support matrices,
The dry reagent visually distinguishes the amniotic fluid from the urine by causing a chemical reaction with a plurality of substances in the woman's secretions,
The dry diagnostic apparatus in which the dry reagent in each of the at least two support matrices can react with different substances in the plurality of substances. The at least two support matrices are: fiber-containing paper, woven and non-woven fabric, synthetic or modified natural polymer, sponge material, cellulose, glass fiber, microporous membrane, wood, (styrene-based copolymer, latex-based, cellulose The dry diagnostic device of claim 1 made from an absorbent material selected from a group of materials such as microporous polymeric materials (such as base or cotton-based matrices). 2. The dry diagnostic apparatus according to claim 1, wherein the dry reagent is capable of reacting with a substance present in the amniotic fluid or the urine, and the substance has a concentration that is significantly higher than the original concentration in the amniotic fluid or the urine. . The dry diagnostic apparatus according to claim 3, wherein the substance includes substances such as creatinine, alkaline phosphatase, total protein, urea, urobilinogen, and blood. The dry diagnostic apparatus according to claim 1, wherein one of the dry reagents in one of the at least two support matrices is capable of reacting with creatinine. One of the support matrices includes two reagent layers, one of the reagent layers includes a creatinine sensitive dye fixed to a dye fixative, and the other reagent layer is relatively high in one of the test areas The dry diagnostic apparatus according to claim 5, comprising a buffering agent capable of maintaining a pH value. 4. The dry diagnostic device of claim 3, wherein one of the carrier matrices includes a single layer comprising a creatinine sensitive dye, a buffer and a dye fixative. The creatinine sensitive dye is 3′5′-dinitrobenzoic acid, 2′4′-dinitrobenzoic acid, 3′5′-dinitrobenzotrifluoride, 3′5′-dinitrobenzamide, 3′5′-dinitrobenzoylphenyl. The dry diagnostic apparatus according to claim 6 or 7, wherein the dry diagnostic apparatus is selected from the group of dinitro derivatives such as glycine and 3,5-dinitrohydroxyphenylpropionic acid. The dye fixing agent is selected from polymerized quaternary ammonium cations (quats) such as polydiallyldimethylammonium chloride, polymonoallyltrimethylammonium chloride, polytrimethylaminoethyl methacrylate chloride, polyvinylbenzyltrimethylammonium chloride, and polyvinylmethylpyridine chloride. The dry diagnostic apparatus according to claim 6 or 7. The dry diagnostic apparatus according to claim 6 or 7, wherein the creatinine sensitive dye and the dye fixing agent are buffered so as to maintain a stable pH within a range of about 9 to 13.5. 7. The buffering agent is selected from the group of sodium metasilicate, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydroxide-potassium chloride, potassium carbonate, glycine-sodium hydroxide, and sodium borate. Or the dry-type diagnostic apparatus of 7. The dry diagnostic apparatus according to claim 1, wherein one of the dry reagents in one of the support matrices is capable of reacting with total protein. One of the dry reagents is 3 ′, 3 ″, 5 ′, 5 ″ -tetrabromophenolsulfonephthalein, Coomassie brilliant blue, first green, light green, pyrogallol sulfonephthalein (pyrogallol red), pyrocatechol sulfone. Phthalein (pyrocatechol violet), 3 ', 3 "-dibromo-5', 5" -dichlorophenolsulfonephthalein, fuchsic acid, 2,4-dinitro-1-naphthol (Martus yellow) Phloxine B, Congo red The dry diagnostic apparatus according to claim 12, wherein the dye is selected from the group consisting of ethyl orange and methyl orange. The dry diagnostic apparatus according to claim 13, further comprising a buffer such as potassium citrate, potassium chloride, potassium sulfate, potassium iodate, or potassium phosphate. The dry diagnostic apparatus according to claim 13, further comprising a metal ion selected from copper, lead, zinc, and silver. The dry diagnostic apparatus of claim 1, wherein one of the dry reagents in one of the support matrices is capable of reacting with alkaline phosphatase. One of the dry reagents is an alkaline phosphate substrate selected from p-nitrophenyl phosphate, indoxyl phosphate, 4-methylumbelliferyl phosphate and alpha-naphthyl phosphate The dry diagnostic apparatus according to claim 16. The dry diagnostic apparatus according to claim 1, wherein one of the dry reagents is a pH sensitive reagent. The dry diagnostic apparatus according to claim 1, wherein the dry reagent does not exit the support matrix. The dry diagnostic apparatus according to claim 1, wherein the at least two carrier matrices are covered with a protective layer. The dry diagnostic apparatus according to claim 20, wherein the protective layer is transparent. 21. The dry diagnostic apparatus of claim 20, wherein the protective layer is made of a thin "one-way structure" membrane that is permeable to liquid flowing through the at least two support matrices and prevents the reagent from flowing out of the apparatus. . The dry diagnostic apparatus according to claim 1, wherein an adhesive backing layer is provided below the base layer. 24. The dry diagnostic apparatus according to claim 23, wherein the adhesive backing layer is provided in proximity to an external protective layer. The dry diagnostic apparatus according to claim 1, wherein the at least two supported reagent matrices are arranged in substantially parallel rows. The dry diagnostic apparatus according to claim 1, wherein the at least two reagent carrying matrices are arranged concentrically. 2. The diagnostic device can be attached when at least two of the test areas face a vaginal canal of a female animal by surrounding the at least two reagent-bearing matrices with an adhesive material. Dry diagnostic equipment.

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