CN107779494A - A kind of kit for detecting infective vaginitis and preparation method thereof - Google Patents

A kind of kit for detecting infective vaginitis and preparation method thereof Download PDF

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Publication number
CN107779494A
CN107779494A CN201711064335.XA CN201711064335A CN107779494A CN 107779494 A CN107779494 A CN 107779494A CN 201711064335 A CN201711064335 A CN 201711064335A CN 107779494 A CN107779494 A CN 107779494A
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pad
reacting
substrate solution
substrate
added dropwise
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CN107779494B (en
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凌燕
王珊珊
吴海燕
张丽梅
任美玲
李�瑞
桂春爽
李小双
刘希
刘金玲
董婷婷
王维
万松平
康培玉
王加义
焦国宾
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BEIJING ZHONGSHENG JINYU DIAGNOSIS TECHNOLOGY Co Ltd
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BEIJING ZHONGSHENG JINYU DIAGNOSIS TECHNOLOGY Co Ltd
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    • G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
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Abstract

The present invention provides a kind of kit for detecting infective vaginitis.The kit includes dry chemical reaction unit, sample diluting liquid and at least one developer：Wherein reaction unit has the reacting hole of at least one following index of detection：Hydrogen peroxide, neuraminidase, leukocyte esterase, β glycuronidases, coagulase, lactic acid, acetyl glucosaminidase, beta lactamase, proline aminopeptidase, alanine aminopeptidase；Contain TritonX 100 and natural gum in sample diluting liquid；Developer is made up of at least one of solid blue B, fast red B, solid blue BB, Fast Blue RR, solid purple B or fast red TR.Reagent kit product of the present invention can assess person under inspection's genital tract micro-ecological environment, cleannes and bacterial vaginosis BV pathogenic bacteria shape simultaneously, and its is easy to operate, quick, cost is cheap.

Description

A kind of kit for detecting infective vaginitis and preparation method thereof
Technical field
The invention belongs to medical test determination techniques field, specifically, is related to a kind of examination for detecting infective vaginitis Agent box and preparation method thereof.
Background technology
Infective vaginitis/disease is gynaecology's common disease, when pathogen intrusion vagina, sticks vagina under certain conditions Film produces inflammatory activity and forms vaginitis, and it is most common female sex organ inflammation, and each age level can suffer from Suffer from.The incidence of disease accounts for the 1/3 of outpatients of gynecology, and clinically about 50%-60% vaginitis is mixed infection.
Vagina mixed infection refers to the infection caused by 2 kinds or two or more pathogen, if relying on microscopy or reagent merely Box, detected just for certain colpitis or disease, clinically more than 50% vaginitis/disease, which will occur, fails to pinpoint a disease in diagnosis, by mistake Situation about examining.
Therefore, in order to reduce diagnosis during occur fail to pinpoint a disease in diagnosis, Misdiagnosis, being directed to the detection of mixed infection is Very necessary, this can not only improve the correct diagnosis and treatment rate of diagnosis and treatment of patient, while can help the administration suggestion of successive treatment, carry The rehabilitation rate of high patient, reduces ill recurrence rate.As can be seen here, tried for the joint-detection of infective vaginitis/sick multi objective Agent box is very necessary.
The content of the invention
It is an object of the invention to provide a kind of kit for detecting infective vaginitis and preparation method thereof.
In order to realize the object of the invention, the kit of detection infective vaginitis provided by the invention, it includes：
1) dry chemical reaction unit
The reaction unit has the reacting hole of at least one following index of detection：It is hydrogen peroxide, neuraminidase, white thin Born of the same parents' esterase, GRD beta-glucuronidase, coagulase, lactic acid, acetyl glucosaminidase, beta-lactamase, proline aminopeptidase, Alanine aminopeptidase；
2) sample diluting liquid；
3) at least one developer.
Foregoing kit, the reaction substrate of the reaction unit are made up of ABS engineering plastics, if wherein substrate is provided with Dry shape, hole of the same size, the reacting hole of respectively above-mentioned each index, equipped with corresponding reaction in each reacting hole Pad.
Preferably, have 10 a diameter of 0.4mm circular hole on the substrate of the reaction unit, respectively as hydrogen peroxide, Neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, lactic acid, acetyl glucosaminidase, beta-lactam Enzyme, proline aminopeptidase, the reacting hole of alanine aminopeptidase.
The hydroperoxidation hole of the reaction unit is coated with horseradish peroxidase, 3- methyl -2-[4-morpholinodithio quinoline Ketone hydrazone hydrochloride hydrate and 3- (N- ethyl -3- toluidines) -2- hydroxy-propanesulfonic acid sodium salts.
The neuraminidase reacting hole of the reaction unit is coated with the chloro- 3- indoles-N-acetyl-neuraminates of the bromo- 4- of 5- Salt, its corresponding developer A are solid purple B or fast red TR.
The leukocyte esterase reacting hole of the reaction unit is coated with the chloro- 3- indole-acetic acids salt of the bromo- 4- of 5-.
The GRD beta-glucuronidase reacting hole of the reaction unit is coated with the chloro- 3- indoles-beta-glucuronic acids of the bromo- 4- of 5- Salt.
The coagulase reacting hole of the reaction unit is coated with glycyl arginine-beta-naphthylamine, and its corresponding developer B is Gu blue B, fast red B, solid blue BB or Fast Blue RR, preferably solid blue B.
The lactic acid reacting hole of the reaction unit is coated with D-lactic acid dehydrogenase, diaphorase and glutamic-pyruvic transaminase.
The acetyl glucosaminidase reacting hole of the reaction unit is coated with the chloro- 3- indyls-N- acetyl of the bromo- 4- of 5- Base-β-D- glucosaminides, its corresponding developer A are solid purple B or fast red TR.
The beta-lactamase reacting hole of the reaction unit is coated with Nitrocefin.
The proline aminopeptidase reacting hole of the reaction unit is coated with L-PROLINE -4- methoxyl groups-beta-naphthylamine hydrochloride, Its corresponding developer B is solid blue B, fast red B, admittedly blue BB or Fast Blue RR, preferably solid blue B.
The alanine aminopeptidase reacting hole of the reaction unit is coated with glycyl arginine β naphthylamines, colour developing corresponding to it Agent B is solid blue B, fast red B, admittedly blue BB or Fast Blue RR, preferably solid blue B.
Foregoing kit, contain TritonX-100 and natural gum in the sample diluting liquid.
The kit of the detection infective vaginitis of the present invention, can be prepared as follows to obtain：
(1) preparation of substrate is reacted：
Reaction substrate be made up of ABS engineering plastics, wherein by have made of ABS engineering plastics on substrate 10 it is a diameter of 0.4mm circular hole, hydrogen peroxide, neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, breast are corresponded to respectively Acid, acetyl glucosaminidase, beta-lactamase, proline aminopeptidase, alanine aminopeptidase hole, are filled out in each reacting hole Enter corresponding reacting pad carrier, used carrier is filter paper, glass fibre or chromatographic paper；
(2) preparation of hydroperoxidation pad, enzyme pad and substrate pad are stacked with successively from hole is bottom up：
The enzyme pad is added dropwise by enzyme solutions and is prepared on carrier；With 0.001-1g 3- methyl -2-[4-morpholinodithio quinoline ketone Hydrazone hydrochloride hydrate, dissolved with 10mL water, the horseradish peroxidase for then adding 10000-10000U is made；
The substrate pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with 100-300mmol/L pH 7.0-8.0 BK- citrate buffer solutions are solvent, add 0.001-1g3- (N- ethyl -3- toluidines) third sulphur of -2- hydroxyls Acid sodium-salt is made；
(3) preparation of neuraminidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with NaClO containing 1-10mol/L 100-300mmol/L pH5.0-6.0 acetate buffer be solvent, add the chloro- 3- indoles of the bromo- 4- of 0.001-1g 5-- N-acetyl-neuraminate salt is made；
(4) preparation of leukocyte esterase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with the sodium sulphate containing 2wt% 100-1000mmol/L pH 6.0-7.0 phosphate buffer is solvent, the chloro- 3- indoles of the addition bromo- 4- of 0.001-1g 5-- Acetate is made；
(5) preparation of GRD beta-glucuronidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with second containing 10-100mmol/L two The 100-300mmol/L pH 6.0-7.0 of alcohol phosphate buffer is solvent, adds the chloro- 3- Yin of the bromo- 4- of 0.001-1g 5- Diindyl-beta-glucuronic acid salt is made；
(6) preparation of coagulase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with 100-300mmol/L pH 8.0-9.0 Tris-HCl buffer solutions are solvent, add 0.001-1g glycyl arginine-beta-naphthylamine and are made；
(7) preparation of lactic acid reacting pad, substrate pad and enzyme pad are stacked with successively from hole is bottom up：
The substrate pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is molten by 500 μ L dimethyl sulfoxide (DMSO)s 0.001-0.1g NBT (NBT) are solved, after adding 2mL saturation beta-schardinger dextrin solution, add 2.5mL lactic acid Developer protects liquid and 0.001-0.1g NAD (NADH) to be made；
The enzyme pad is added dropwise by enzyme solutions and is prepared on carrier, and enzyme solutions add using 5mL enzyme protections liquid as solvent 5KU D-lactic acid dehydrogenases, stir to being completely dissolved, add 10-100mg diaphorase and 10-100mg glutamic-pyruvic transaminase is made；
The lactic acid developer protection liquid of above-mentioned substrate solution is added by pH6-7 0.1-1mol/L Tris-HCl buffer solutions 10-100g Dextran T 70s and 10-100g inositols are made；
Above-mentioned enzyme protection liquid adds 1-10g sucrose, 1-10g D- by pH7-8 500-600mmol/L glycylglycine buffer solutions Trehalose, 1-10g bovine serum albumin(BSA)s, 1-10g inositols and 100-1000 μ L Tween-20s are made；
(8) preparation of acetyl glucosaminidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is with pH 5.0-6.0 500- 600mmol/L citric acid phosphoric acid disodium hydrogens cushioning liquid is solvent, adds the chloro- 3- indyls-N- second of the bromo- 4- of 0.001-1g 5- Acyl-beta-D- glucosaminides, while add Dextran T 70 and L-Histidine is made；Dextran T 70 and L- in substrate solution Histidine final concentration is 0.005%-50%, the final concentration of 0.005%-50% of L-Histidine；
(9) preparation of beta-lactamase reacting pad, substrate pad and cushion pad are stacked with successively from hole is bottom up：
The substrate pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is made using isopropanol as solvent 0.001-0.01g/mL Nitrocefins, 0.005%-50% methylcellulose, 10% polyvinylpyrrolidone, the Guangs of 1%L- half The mixed liquor of propylhomoserin and 50mmol/L citric acids is as substrate solution；
The cushion pad is added dropwise by cushioning liquid and is prepared on carrier, and cushioning liquid is 200-600mmol/L pH 7.0-8.0 phosphate buffer；
(10) preparation of proline aminopeptidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is with pH 8.0-9.0 400- 500mmol/L Tris-HCl is buffer solution, and 10-40mg/mL L-PROLINEs -4- methoxyl groups-beta-naphthylamine, 0.005%- is made 50%TritonX-100,1% Dextran T 70 and 1%D- trehaloses mixed liquor are as substrate solution；
(11) preparation of alanine aminopeptidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is with 500-1000mmol/L PH8.0-9.0 Tris-HCl buffer solutions are solvent, be made 0.001-0.01g/mL ALANINEs -4- methoxyl groups-beta-naphthylamine, The mixed liquor of 0.005%-50% Dextran T 70s, 1%D- trehaloses and 10%TritonX-100 is as substrate solution；
(12) reacting pad prepared by step (2)-(11) is inserted in corresponding reacting hole, be compacted with stamping machine, on hole Protection foil film is pasted, is fitted into aluminium foil bag, produces reaction unit, in 2-8 DEG C of preservation；
(13) preparation of sample diluting liquid：
The sample diluting liquid contains 0.9%NaCL, 10mmol/L magnesium chloride, 2mmol/L zinc chloride, 4mmol/L chlorinations Manganese and 10 ± 2mmol/L MES, pH6.5 ± 0.2；
(14) preparation of developer：
Using polyoxyethylene laurel ether as solvent, the solid purple B or fast red TR for preparing mass percentage concentration 0.05~0.3% are molten Liquid；
Using polyoxyethylene laurel ether as solvent, the solid blue B, fast red B, solid indigo plant of mass percentage concentration 0.05~0.3% are prepared BB or Fast Blue RR solution.
In the specific embodiment of the present invention, the preparation method of the detection kit is as follows：
(1) preparation of substrate is reacted：
Reaction substrate be made up of ABS engineering plastics, wherein by have made of ABS engineering plastics on substrate 10 it is a diameter of 0.4mm circular hole, hydrogen peroxide, neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, breast are corresponded to respectively Acid, acetyl glucosaminidase, beta-lactamase, proline aminopeptidase, alanine aminopeptidase hole, are filled out in each reacting hole Enter corresponding reacting pad carrier, used carrier is filter paper, glass fibre or chromatographic paper；
(2) preparation of hydroperoxidation pad, enzyme pad and substrate pad are stacked with successively from hole is bottom up：
Enzyme pad is added dropwise by the μ L of enzyme solutions 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Enzyme solutions 10mL water dissolves 0.011g 3- methyl-2-benzothiazolinone hydrazone hydrochloride hydrates, adds 15000UU horseradish peroxidase is made；
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution is solvent with the 200mmo/L of 10mL pH 6.8 BK- citrate buffer solutions, adds 0.140g 3- (N- ethyl -3- toluidines) -2- hydroxy-propanesulfonic acid sodium salts are made；
(3) preparation of neuraminidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution adds 0.05g with the 200mmol/L pH5.5 of the NaClO containing 2mol/L acetate buffer 10mL The chloro- 3- indoles-N-acetyl-neuraminate salt of the bromo- 4- of 5- is made；
(4) preparation of leukocyte esterase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution adds using the 300mmol/L pH8.3 of the sodium sulphate containing 2wt% phosphate buffer 1 0mL as solvent The chloro- 3- indole-acetic acids salt of the bromo- 4- of 0.05g 5- is made；
(5) preparation of GRD beta-glucuronidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution using the 200mmol/L pH7.0 of the ethylene glycol containing 50mmol/L phosphate buffer 1 0mL as solvent, The chloro- 3- indoles-beta-glucuronic acid salt of the bromo- 4- of 0.05g 5- is added to be made；
(6) preparation of coagulase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution with 200mmol/L pH 8.5 Tris-HCl buffer solution 10mL, add 0.05g glycyl arginine- Beta-naphthylamine is made；
(7) preparation of lactic acid reacting pad, substrate pad and enzyme pad are stacked with successively from hole is bottom up：
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution after adding 2mL saturation beta-schardinger dextrin solution, is added by 500 μ L dmso solution 0.01g NBT Enter 2.5mL lactic acid developer protection liquid and 0.065g NAD are made；
Lactic acid developer protection liquid is the 0.8mol/L Tris-HCl buffer solutions of pH 6.5 with 80mL, adds 16g Portugals and gathers Sugared T70 and 12g inositols are made；
Enzyme pad is added dropwise by the μ L of enzyme solutions 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Enzyme solutions add 5KU D-lactic acid dehydrogenases using 5mL enzyme protections liquid as solvent, stir to being completely dissolved, add 76.9mg diaphorase and 35mg glutamic-pyruvic transaminase are made；
Enzyme protection liquid adds 2g sucrose, 2g D- trehaloses, 1g by the 600mmol/L glycylglycine buffer solution 45mL of pH 7.5 Bovine serum albumin(BSA), 3g inositols and 250 μ L Tween-20s are made；
(8) preparation of acetyl glucosaminidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution adds 0.025g 5- with the 500mmol/L citric acid phosphoric acid disodium hydrogen cushioning liquid 5mL of pH 5.0 The chloro- 3- indyls-N- acetyl group-β-D- glucosaminides of bromo- 4-, 0.025g Dextran T 70s, 0.0582g L-Histidine systems Into；
(9) preparation of beta-lactamase reacting pad, substrate pad and cushion pad are stacked with successively from hole is bottom up：
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
10g/mL Nitrocefins, 1% methylcellulose, 10% polyethylene is made using isopropanol as solvent in substrate solution The mixed liquor of pyrrolidones, 1%L- cysteines and 50mmol/L citric acids is as substrate solution；
Cushion pad is added dropwise by the μ L of cushioning liquid 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
The cushion pad is added dropwise by cushioning liquid and is prepared on carrier, cushioning liquid pH7.0,200mmol/L phosphorus Phthalate buffer；
(10) preparation of proline aminopeptidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
20mg/mL L- dried meat ammonia is made using the 500mmol/L of pH 8.0 Tris-HCl 5mL as buffer solution in substrate solution Acid -4- methoxyl groups-beta-naphthylamine, 10%TritonX-100, the mixed liquor of 1% Dextran T 70 and 1%D- trehaloses are as substrate Solution；
(11) preparation of alanine aminopeptidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
3.5g/L ALANINEs -4- is made with the 500mmol/L of pH 8.5 Tris-HCl buffer solution 5mL in substrate solution Methoxyl group-beta-naphthylamine, 1% Dextran T 70,1%D- trehaloses and 10%TritonX-100 mixed liquor are as substrate solution.
(12) reacting pad prepared by step (2)-(11) is inserted in corresponding reacting hole, be compacted with stamping machine, on hole Protection foil film is pasted, is fitted into aluminium foil bag, produces reaction unit, in 2-8 DEG C of preservation.
(13) preparation of sample diluting liquid：
The sample diluting liquid contains 0.9%NaCL, 10mmol/L magnesium chloride, 2mmol/L zinc chloride, 4mmol/L chlorinations Manganese and 10 ± 2mmol/L MES, pH6.5 ± 0.2.
(14) preparation of developer：
Using polyoxyethylene laurel ether as solvent, the solid purple B or fast red TR for preparing mass percentage concentration 0.05~0.3% are molten Liquid；
Using polyoxyethylene laurel ether as solvent, the solid blue B, fast red B, solid indigo plant of mass percentage concentration 0.05~0.3% are prepared BB or Fast Blue RR solution.
The present invention further provides application of the mentioned reagent box in infective vaginitis are detected.Protected in view of kit The needs deposited, prepared reacting pad must possess at a relatively high stability, final to determine on the basis of lot of experiments The composition and its formula of above-mentioned enzyme liquid, nitrite ion and various substrate solutions so that kit has preferable stability, Neng Gou Promoted in practical application.
Compared with prior art, the present invention has advantages below：
(1) present invention solves the instability problem of liquid reagent, by adding the materials such as activator, enzyme stabilizers, On guarantee reagent pad on the premise of stable reagent, sensitivity and the accuracy of reaction can be rapidly improved；Kit operation letter Just quick, accuracy is high, and suitable for routine clinical detection, detection uses particularly by clinical patient bed, and then instructs sound clinical Medication.
(2) it is accurate to clinically infective vaginitis progress detection kit testing result using the kit of the present invention Rate is up to more than 95%.
(3) kit of the present invention, which avoids, individually observes one-sidedness and disturbing factor caused by single index, using joint Detection more accurate and effective of the detection of index for clinically vagina mixed infection.
(4) reagent kit product of the present invention can use in hospital emergency rooms, primary care and care facilities and patient family. Person under inspection's genital tract micro-ecological environment, cleannes and bacterial vaginosis BV pathogenic bacteria shape can be assessed simultaneously, and its is easy to operate, fast Speed, cost are cheap.
Embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.Unless otherwise specified, embodiment In the conventional meanses that are well known to those skilled in the art of used technological means, raw materials used is commercial goods.
The percentage sign " % " being related in the present invention, if not specified, refers to mass percent；But the percentage of solution Than unless otherwise specified, referring to the grams containing solute in 100mL solution.
A kind of preparation of the dry chemical detection kit of 1 infective vaginitis of embodiment
The infective vaginitis detection kit of the present embodiment includes：1. dry chemical reaction unit：With detection peroxidating Hydrogen, neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, lactic acid, acetyl glucosaminidase, β-interior acyl Amine enzyme, proline aminopeptidase, the reaction substrate of alanine aminopeptidase index reacting hole；2. vaginal fluid dilution；3. develop the color Agent A and developer B.
Reaction unit is prepared as follows obtaining：
(1) preparation of substrate is reacted：
Reaction substrate be made up of ABS engineering plastics, wherein by have made of ABS engineering plastics on substrate 10 it is a diameter of 0.4mm circular hole, hydrogen peroxide, neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, breast are corresponded to respectively Acid, acetyl glucosaminidase, beta-lactamase, proline aminopeptidase, alanine aminopeptidase hole, are filled out in each reacting hole Enter corresponding reacting pad carrier, used carrier is filter paper, glass fibre or chromatographic paper；
(2) preparation of hydroperoxidation pad, enzyme pad and substrate pad are stacked with successively from hole is bottom up：
Enzyme pad is added dropwise by the μ L of enzyme solutions 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Enzyme solutions 10mL water dissolves 0.011g 3- methyl-2-benzothiazolinone hydrazone hydrochloride hydrates, adds 15000UU horseradish peroxidase is made；
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution is solvent with the 200mmo/L of 10mL pH 6.8 BK- citrate buffer solutions, adds 0.140g 3- (N- ethyl -3- toluidines) -2- hydroxy-propanesulfonic acid sodium salts are made；
(3) preparation of neuraminidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution adds 0.05g with the 200mmol/L pH5.5 of the NaClO containing 2mol/L acetate buffer 10mL The chloro- 3- indoles-N-acetyl-neuraminate salt of the bromo- 4- of 5- is made；
(4) preparation of leukocyte esterase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution adds using the 300mmol/L pH8.3 of the sodium sulphate containing 2wt% phosphate buffer 1 0mL as solvent The chloro- 3- indole-acetic acids salt of the bromo- 4- of 0.05g 5- is made；
(5) preparation of GRD beta-glucuronidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution using the 200mmol/L pH7.0 of the ethylene glycol containing 50mmol/L phosphate buffer 1 0mL as solvent, The chloro- 3- indoles-beta-glucuronic acid salt of the bromo- 4- of 0.05g 5- is added to be made；
(6) preparation of coagulase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution with 200mmol/L pH 8.5 Tris-HCl buffer solution 10mL, add 0.05g glycyl arginine- Beta-naphthylamine is made；
(7) preparation of lactic acid reacting pad, substrate pad and enzyme pad are stacked with successively from hole is bottom up：
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution after adding 2mL saturation beta-schardinger dextrin solution, is added by 500 μ L dmso solution 0.01g NBT Enter 2.5mL lactic acid developer protection liquid and 0.065g NAD are made；
Lactic acid developer protection liquid is the 0.8mol/L Tris-HCl buffer solutions of pH 6.5 with 80mL, adds 16g Portugals and gathers Sugared T70 and 12g inositols are made；
Enzyme pad is added dropwise by the μ L of enzyme solutions 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Enzyme solutions add 5KU D-lactic acid dehydrogenases using 5mL enzyme protections liquid as solvent, stir to being completely dissolved, add 76.9mg diaphorase and 35mg glutamic-pyruvic transaminase are made；
Enzyme protection liquid adds 2g sucrose, 2g D- trehaloses, 1g by the 600mmol/L glycylglycine buffer solution 45mL of pH 7.5 Bovine serum albumin(BSA), 3g inositols and 250 μ L Tween-20s are made；
(8) preparation of acetyl glucosaminidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
Substrate solution adds 0.025g 5- with the 500mmol/L citric acid phosphoric acid disodium hydrogen cushioning liquid 5mL of pH 5.0 The chloro- 3- indyls-N- acetyl group-β-D- glucosaminides of bromo- 4-, 0.025g Dextran T 70s, 0.0582g L-Histidine systems Into；
(9) preparation of beta-lactamase reacting pad, substrate pad and cushion pad are stacked with successively from hole is bottom up：
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
10g/mL Nitrocefins, 1% methylcellulose, 10% polyethylene is made using isopropanol as solvent in substrate solution The mixed liquor of pyrrolidones, 1%L- cysteines and 50mmol/L citric acids is as substrate solution；
Cushion pad is added dropwise by the μ L of cushioning liquid 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
The cushion pad is added dropwise by cushioning liquid and is prepared on carrier, cushioning liquid pH7.0,200mmol/L phosphorus Phthalate buffer；
(10) preparation of proline aminopeptidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
20mg/mL L- dried meat ammonia is made using the 500mmol/L of pH 8.0 Tris-HCl 5mL as buffer solution in substrate solution Acid -4- methoxyl groups-beta-naphthylamine, 10%TritonX-100, the mixed liquor of 1% Dextran T 70 and 1%D- trehaloses are as substrate Solution；
(11) preparation of alanine aminopeptidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs dry 4 It is made, is sealed at 4 DEG C after hour；
3.5g/L ALANINEs -4- is made with the 500mmol/L of pH 8.5 Tris-HCl buffer solution 5mL in substrate solution Methoxyl group-beta-naphthylamine, 1% Dextran T 70,1%D- trehaloses and 10%TritonX-100 mixed liquor are as substrate solution.
Reacting pad prepared by above-mentioned steps (2)-(11) is inserted in corresponding reacting hole, is compacted with stamping machine, on hole Protection foil film is pasted, is fitted into aluminium foil bag, produces reaction unit, in 2-8 DEG C of preservation.
The preparation of vaginal fluid dilution (sample diluting liquid)：
The sample diluting liquid contains 0.9%NaCL, 10mmol/L magnesium chloride, 2mmol/L zinc chloride, 4mmol/L chlorinations Manganese and 10 ± 2mmol/L MES, pH6.5 ± 0.2.
The preparation of developer：
Using polyoxyethylene laurel ether as solvent, the solid purple B or fast red TR for preparing mass percentage concentration 0.05~0.3% are molten Liquid；
Using polyoxyethylene laurel ether as solvent, the solid blue B, fast red B, solid indigo plant of mass percentage concentration 0.05~0.3% are prepared BB or Fast Blue RR solution.
The application of the dry chemical detection kit of 2 vaginitis of the present invention of embodiment
The kit of the present invention can both detect to the vaginal fluid sample for clinically suffering from gynaecological imflammation crowd, Also the vaginal fluid sample of healthy population can be detected, can conduct to check whether crowd to be measured suffers from vaginal disease The auxiliary characteristics of diagnosis, antidiastole and the therapeutic evaluation of patient, the backman that can be also evaluated as Healthy People microecology in vaginas Tool.
The specifically used method of kit prepared by the embodiment of the present invention 1 is as follows：
1) when in use, test paper pad (reacting pad) is taken out under storage temperature first, room temperature is equilibrated to and begins to use；
2) dilution is handled：400 μ L diluted secretion are added after taking secretion from posterior fornix with cotton swab；
3) 25 μ L secretion dilutions are taken to be added separately on test paper pad；
4) nitrite ion A is added dropwise in neuraminidase and acetyl glucosaminidase hole；
5) after reacting 7min on dry bath device (it is 45 DEG C to set temperature), coagulase, proline aminopeptidase and phenylalanine ammonia Nitrite ion B is added dropwise in peptase hole, continues to react each hole colour developing result of 3min readings.It will be appreciated by those skilled in the art that according to this The prior art in field, it is 35-50 DEG C in the setting temperature using other cool-bags such as incubator, baking oven, water bath containers etc. Under conditions of react after, can also realize result contrast judge.
The kit can the disposable clinical common colpitis of quick diagnosis, it is micro- to be mainly used in auxiliary diagnosis vagina Environmental health situation, including bacterial vaginosis BV (BV), VVC (VVC), trichomonas vaginitis (TV), Aerobic bacteria property vaginitis (AV).Wherein Gram-negative bacteria, dried meat ammonia be present in the positive prompting vaginal fluid of alanine aminopeptidase The positive prompting BV or VVC of sour aminopeptidase is positive, and the prompting of beta-lactam enzyme positive has beta-lactam antibiotic drug-fast bacteria, acetyl The positive prompting VVC or TV of UNAG is positive, is carried inside the positive prompting vagina of lactic acid with the presence of lactic acid, coagulase positive Show the AV positives, beta-glucosidase enzyme positive prompting AV is positive, and the positive prompting inflammation of leukocyte esterase is present, and neuraminidase is positive Prompt BV positive, the positive prompting microecology in vaginas of hydrogen peroxide is abnormal.Result above is only for reference, need to combine clinical other pathology Indication carries out comprehensive descision.
Reaction result criterion：The aobvious blueness of peroxidating hydrogen holes, neuraminidase hole is not developed the color or yellow, leukocyte esterase Hole is not developed the color, and beta-glucuronidase enzyme hole is not developed the color, coagulase hole is not developed the color or yellow, the aobvious blueness in lactic acid hole, acetylamino Glucuroide does not develop the color or displaing yellow, beta-lactamase hole displaing yellow, Prolyl iminopeptidase does not develop the color or displaing yellow, the third ammonia Sour aminopeptidase does not develop the color or displaing yellow, is negative reaction result；Peroxidating hydrogen holes does not develop the color or aobvious green, neuraminidase hole Displaing amaranth or blueness, the aobvious blueness in leukocyte esterase hole or green, are positive reaction result；The aobvious indigo plant in beta-glucuronidase enzyme hole Color or green, coagulase hole show purple red, crocus, and lactic acid hole is not developed the color, acetyl glucosaminidase hole displaing amaranth, The aobvious red in beta-lactamase hole, aobvious red, the aobvious red of Alanine aminopeptidase of Prolyl iminopeptidase, is positive reaction result.
The kit stable performance of the present invention, shelf-life are 18 months.
Clinically infective vaginitis are detected using the kit of the present invention, kit testing result of the present invention is accurate True rate is up to more than 95%.
Although above the present invention is described in detail with a general description of the specific embodiments, On the basis of the present invention, it can be modified or improved, this will be apparent to those skilled in the art.Cause This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Bibliography
1.Sobel,J.D.,Vaginitis.New England Journal of Medicine,1997.337(26): p.1896.
2.Pheifer,T.A.,et al.,Nonspecific vaginitis:role of Haemophilus vaginalis and treatment with metronidazole.New England Journal of Medicine, 1978.298(26):p.1429.
3. a duckweed, the treatment China Non communicable disease of common vaginal inflammation, 2011.19 (2):p.218-219.
4. the yellow love of the just beautiful and of Gao is military, the antidiastole China microecology of vagina simple infection and multiple infection is miscellaneous Will, 2013.25 (2):p.206-207.
5. Xiao Bing ice, Liu Chaohui, and Liao Qin are put down, Out-patient Clinic of Department of Gynecology difference vaginal symptoms clients microecology in vaginas situation is adjusted Look into China journal of obstetrics and gynecology, 2009.44 (1):p.6-8.

Claims (7)

1. a kind of kit for detecting infective vaginitis, it is characterised in that it includes：
1) dry chemical reaction unit
The reaction unit has the reacting hole of at least one following index of detection：Hydrogen peroxide, neuraminidase, leucocyte ester Enzyme, GRD beta-glucuronidase, coagulase, lactic acid, acetyl glucosaminidase, beta-lactamase, proline aminopeptidase, the third ammonia Sour aminopeptidase；
2) sample diluting liquid；
3) at least one developer.
2. kit according to claim 1, it is characterised in that the reaction substrate of the reaction unit is by ABS engineering plastics Material is made, and wherein substrate is provided with several shapes, hole of the same size, the reacting hole of respectively above-mentioned each index, each Corresponding reacting pad is housed in reacting hole.
3. kit according to claim 2, it is characterised in that have on the substrate of the reaction unit 10 it is a diameter of 0.4mm circular hole.
4. kit according to claim 2, it is characterised in that the hydroperoxidation hole of the reaction unit is coated with Horseradish peroxidase, 3- methyl-2-benzothiazolinone hydrazones hydrochloride hydrate and 3- (N- ethyl -3- toluidines) - 2- hydroxy-propanesulfonic acid sodium salts；
The neuraminidase reacting hole of the reaction unit is coated with the chloro- 3- indoles-N-acetyl-neuraminate salt of the bromo- 4- of 5-, its Corresponding developer A is solid purple B or fast red TR；
The leukocyte esterase reacting hole of the reaction unit is coated with the chloro- 3- indole-acetic acids salt of the bromo- 4- of 5-；
The GRD beta-glucuronidase reacting hole of the reaction unit is coated with the chloro- 3- indoles-beta-glucuronic acid salt of the bromo- 4- of 5-；
The coagulase reacting hole of the reaction unit is coated with glycyl arginine-beta-naphthylamine, and its corresponding developer B is solid indigo plant B, fast red B, solid blue BB or Fast Blue RR, preferably solid blue B；
The lactic acid reacting hole of the reaction unit is coated with D-lactic acid dehydrogenase, diaphorase and glutamic-pyruvic transaminase；
The acetyl glucosaminidase reacting hole of the reaction unit be coated with the chloro- 3- indyls-N- acetyl group of the bromo- 4- of 5-- β-D- glucosaminides, its corresponding developer A are solid purple B or fast red TR；
The beta-lactamase reacting hole of the reaction unit is coated with Nitrocefin；
The proline aminopeptidase reacting hole of the reaction unit is coated with L-PROLINE -4- methoxyl groups-beta-naphthylamine hydrochloride, and its is right The developer B answered is solid blue B, fast red B, admittedly blue BB or Fast Blue RR, preferably solid blue B；
The alanine aminopeptidase reacting hole of the reaction unit is coated with glycyl arginine β naphthylamines, and its corresponding developer B is Gu blue B, fast red B, solid blue BB or Fast Blue RR, preferably solid blue B.
5. according to the kit described in claim any one of 1-4, it is characterised in that contain in the sample diluting liquid TritonX-100 and natural gum.
6. the preparation method of any one of the claim 1-5 kits, it is characterised in that comprise the following steps：
(1) preparation of substrate is reacted：
Reaction substrate is made up of ABS engineering plastics, wherein by there is 10 a diameter of 0.4mm made of ABS engineering plastics on substrate Circular hole, correspond to hydrogen peroxide, neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, lactic acid, acetyl respectively UNAG, beta-lactamase, proline aminopeptidase, alanine aminopeptidase hole, inserted in each reacting hole corresponding Reacting pad carrier, used carrier is filter paper, glass fibre or chromatographic paper；
(2) preparation of hydroperoxidation pad, enzyme pad and substrate pad are stacked with successively from hole is bottom up：
The enzyme pad is added dropwise by enzyme solutions and is prepared on carrier；With 0.001-1g 3- methyl-2-benzothiazolinone hydrazone salt Hydrochloride hydrates, dissolved with 10mL water, the horseradish peroxidase for then adding 10000-10000U is made；
The substrate pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with 100-300mmol/L pH 7.0- 8.0 BK- citrate buffer solutions are solvent, add 0.001-1g 3- (N- ethyl -3- toluidines) -2- hydroxy-propanesulfonic acids Sodium salt is made；
(3) preparation of neuraminidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with the NaClO's containing 1-10mol/L 100-300mmol/L pH5.0-6.0 acetate buffer is solvent, adds the chloro- 3- indoles-N- of the bromo- 4- of 0.001-1g 5- N acetylneuraminic acid n salt is made；
(4) preparation of leukocyte esterase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with the 100- of the sodium sulphate containing 2wt% 1000mmol/L pH 6.0-7.0 phosphate buffer is solvent, adds the chloro- 3- indole-acetic acids of the bromo- 4- of 0.001-1g 5- Salt is made；
(5) preparation of GRD beta-glucuronidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with the ethylene glycol containing 10-100mmol/L 100-300mmol/L pH 6.0-7.0 phosphate buffer is solvent, the chloro- 3- indoles-β of the addition bromo- 4- of 0.001-1g 5-- Glucuronate salt is made；
(6) preparation of coagulase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier；Substrate solution is with 100-300mmol/L pH 8.0- 9.0 Tris-HCl buffer solutions are solvent, add 0.001-1g glycyl arginine-beta-naphthylamine and are made；
(7) preparation of lactic acid reacting pad, substrate pad and enzyme pad are stacked with successively from hole is bottom up：
The substrate pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is by 500 μ L dmso solutions 0.001-0.1g NBT, after adding 2mL saturation beta-schardinger dextrin solution, add 2.5mL lactic acid developer protection liquid and 0.001- 0.1g NAD are made；
The enzyme pad is added dropwise by enzyme solutions and is prepared on carrier, and enzyme solutions add 5KU D- using 5mL enzyme protections liquid as solvent Lactic dehydrogenase, stir to being completely dissolved, add 10-100mg diaphorase and 10-100mg glutamic-pyruvic transaminase is made；
The lactic acid developer protection liquid of above-mentioned substrate solution adds 10- by pH6-7 0.1-1mol/L Tris-HCl buffer solutions 100g Dextran T 70s and 10-100g inositols are made；
Above-mentioned enzyme protection liquid adds 1-10g sucrose, 1-10g D- marine algas by pH7-8 500-600mmol/L glycylglycine buffer solutions Sugar, 1-10g bovine serum albumin(BSA)s, 1-10g inositols and 100-1000 μ L Tween-20s are made；
(8) preparation of acetyl glucosaminidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is with pH5.0-6.0 500-600mmol/ L citric acid phosphoric acid disodium hydrogens cushioning liquid is solvent, adds the chloro- 3- indyls-N- acetyl group-β-D- of the bromo- 4- of 0.001-1g 5- Glucosaminide, while add Dextran T 70 and L-Histidine is made；Dextran T 70 and L-Histidine are whole in substrate solution Concentration is 0.005%-50%, the final concentration of 0.005%-50% of L-Histidine；
(9) preparation of beta-lactamase reacting pad, substrate pad and cushion pad are stacked with successively from hole is bottom up：
The substrate pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is made using isopropanol as solvent 0.001-0.01g/mL Nitrocefins, 0.005%-50% methylcellulose, 10% polyvinylpyrrolidone, the Guangs of 1%L- half The mixed liquor of propylhomoserin and 50mmol/L citric acids is as substrate solution；
The cushion pad is added dropwise by cushioning liquid and is prepared on carrier, and cushioning liquid is 200-600mmol/L pH 7.0- 8.0 phosphate buffer；
(10) preparation of proline aminopeptidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is with pH8.0-9.0 400-500mmol/ L Tris-HCl is buffer solution, and 10-40mg/mL L-PROLINEs -4- methoxyl groups-beta-naphthylamine, 0.005%-50% is made TritonX-100,1% Dextran T 70 and 1%D- trehaloses mixed liquor are as substrate solution；
(11) preparation of alanine aminopeptidase reacting pad：
The reacting pad is added dropwise by substrate solution and is prepared on carrier, and substrate solution is with 500-1000mmol/L pH8.0- 9.0 Tris-HCl buffer solutions are solvent, be made 0.001-0.01g/mL ALANINEs -4- methoxyl groups-beta-naphthylamine, The mixed liquor of 0.005%-50% Dextran T 70s, 1%D- trehaloses and 10%TritonX-100 is as substrate solution；
(12) reacting pad prepared by step (2)-(11) is inserted in corresponding reacting hole, is compacted with stamping machine, is pasted on hole Protection foil film, is fitted into aluminium foil bag, produces reaction unit, in 2-8 DEG C of preservation；
(13) preparation of sample diluting liquid：
The sample diluting liquid contain 0.9%NaCL, 10mmol/L magnesium chloride, 2mmol/L zinc chloride, 4mmol/L manganese chlorides and 10 ± 2mmol/L MES, pH6.5 ± 0.2；
(14) preparation of developer：
Using polyoxyethylene laurel ether as solvent, solid purple B or fast red the TR solution of mass percentage concentration 0.05~0.3% is prepared；
Using polyoxyethylene laurel ether as solvent, prepare the solid blue B of mass percentage concentration 0.05~0.3%, fast red B, solid blue BB or Fast Blue RR solution.
7. the preparation method described in claim 6, it is characterised in that comprise the following steps：
(1) preparation of substrate is reacted：
Reaction substrate is made up of ABS engineering plastics, wherein by there is 10 a diameter of 0.4mm made of ABS engineering plastics on substrate Circular hole, correspond to hydrogen peroxide, neuraminidase, leukocyte esterase, GRD beta-glucuronidase, coagulase, lactic acid, acetyl respectively UNAG, beta-lactamase, proline aminopeptidase, alanine aminopeptidase hole, inserted in each reacting hole corresponding Reacting pad carrier, used carrier is filter paper, glass fibre or chromatographic paper；
(2) preparation of hydroperoxidation pad, enzyme pad and substrate pad are stacked with successively from hole is bottom up：
Enzyme pad is added dropwise by the μ L of enzyme solutions 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are made after drying 4 hours , it is sealed at 4 DEG C；
Enzyme solutions 10mL water dissolves 0.011g 3- methyl-2-benzothiazolinone hydrazone hydrochloride hydrates, adds 15000UU Horseradish peroxidase be made；
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution is solvent with the 200mmo/L of 10mL pH 6.8 BK- citrate buffer solutions, adds 0.140g 3- (N- second Base -3- toluidines) -2- hydroxy-propanesulfonic acid sodium salts are made；
(3) preparation of neuraminidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution adds 0.05g 5- with the 200mmol/L pH5.5 of the NaClO containing 2mol/L acetate buffer 10mL The bromo- chloro- 3- indoles-N-acetyl-neuraminate salt of 4- is made；
(4) preparation of leukocyte esterase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution adds using the 300mmol/L pH8.3 of the sodium sulphate containing 2wt% phosphate buffer 1 0mL as solvent The chloro- 3- indole-acetic acids salt of the bromo- 4- of 0.05g 5- is made；
(5) preparation of GRD beta-glucuronidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution adds using the 200mmol/L pH7.0 of the ethylene glycol containing 50mmol/L phosphate buffer 1 0mL as solvent The chloro- 3- indoles-beta-glucuronic acid salt of the bromo- 4- of 0.05g 5- is made；
(6) preparation of coagulase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution adds 0.05g glycyl arginine-β-naphthalene with 200mmol/L pH 8.5 Tris-HCl buffer solution 10mL Amine is made；
(7) preparation of lactic acid reacting pad, substrate pad and enzyme pad are stacked with successively from hole is bottom up：
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution after adding 2mL saturation beta-schardinger dextrin solution, is added by 500 μ L dmso solution 0.01g NBT 2.5mL lactic acid developer protects liquid and 0.065g NAD to be made；
Lactic acid developer protection liquid is the 0.8mol/L Tris-HCl buffer solutions of pH 6.5 with 80mL, adds 16g Dextran T 70s And 12g inositols are made；
Enzyme pad is added dropwise by the μ L of enzyme solutions 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are made after drying 4 hours , it is sealed at 4 DEG C；
Enzyme solutions add 5KU D-lactic acid dehydrogenases using 5mL enzyme protections liquid as solvent, stir to being completely dissolved, add 76.9mg diaphorase and 35mg glutamic-pyruvic transaminase are made；
Enzyme protection liquid adds 2g sucrose, 2g D- trehaloses, 1g ox bloods by the 600mmol/L glycylglycine buffer solution 45mL of pH 7.5 Pure albumen, 3g inositols and 250 μ L Tween-20s are made；
(8) preparation of acetyl glucosaminidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
Substrate solution adds the bromo- 4- of 0.025g 5- with the 500mmol/L citric acid phosphoric acid disodium hydrogen cushioning liquid 5mL of pH 5.0 Chloro- 3- indyls-N- acetyl group-β-D- glucosaminides, 0.025g Dextran T 70s, 0.0582g L-Histidines are made；
(9) preparation of beta-lactamase reacting pad, substrate pad and cushion pad are stacked with successively from hole is bottom up：
Substrate pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
10g/mL Nitrocefins, 1% methylcellulose, 10% polyvinyl pyrrole is made using isopropanol as solvent in substrate solution The mixed liquor of alkanone, 1%L- cysteines and 50mmol/L citric acids is as substrate solution；
Cushion pad is added dropwise by the μ L of cushioning liquid 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
The cushion pad is added dropwise by cushioning liquid and is prepared on carrier, cushioning liquid pH7.0,200mmol/L phosphate Buffer solution；
(10) preparation of proline aminopeptidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
20mg/mL L-PROLINEs -4- is made using the 500mmol/L of pH 8.0 Tris-HCl 5mL as buffer solution in substrate solution Methoxyl group-beta-naphthylamine, 10%TritonX-100, the mixed liquor of 1% Dextran T 70 and 1%D- trehaloses are as substrate solution；
(11) preparation of alanine aminopeptidase reacting pad：
Reacting pad is added dropwise by the μ L of substrate solution 1On chromatographic paper, under the conditions of lucifuge, 25 DEG C of moving airs are dried 4 hours After be made, be sealed at 4 DEG C；
3.5g/LL- alanine -4- methoxies are made with the 500mmol/L of pH 8.5 Tris-HCl buffer solution 5mL in substrate solution Base-beta-naphthylamine, 1% Dextran T 70,1%D- trehaloses and 10%TritonX-100 mixed liquor are as substrate solution.
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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108642131A (en) * 2018-04-17 2018-10-12 山东仕达思生物产业有限公司 A kind of coagulase detection reagent, reacting pad and preparation method thereof and kit
CN108982493A (en) * 2018-09-17 2018-12-11 迪瑞医疗科技股份有限公司 A kind of neuraminidase drying chemical reagent paper and preparation method thereof
CN109265373A (en) * 2018-11-03 2019-01-25 郑州安图生物工程股份有限公司 A kind of amino acids substrate and its preparation method and application
CN109991221A (en) * 2019-03-29 2019-07-09 迪瑞医疗科技股份有限公司 A kind of preparation method of Prolyl iminopeptidase Test paper
CN109991218A (en) * 2019-03-29 2019-07-09 迪瑞医疗科技股份有限公司 A kind of preparation method of coagulase Test paper
CN111690713A (en) * 2020-06-01 2020-09-22 青岛华晶生物技术有限公司 Eight joint inspection kit of vaginitis
CN113030077A (en) * 2021-03-17 2021-06-25 桂林优利特医疗电子有限公司 N-acetylglucosaminidase detection test paper and preparation method thereof
CN113109333A (en) * 2021-04-30 2021-07-13 桂林优利特医疗电子有限公司 Proline aminopeptidase rapid detection test paper and preparation method thereof
CN114317678A (en) * 2021-12-31 2022-04-12 港龙生物技术（深圳）有限公司 Biological paper chip, high-flux multi-connection detection microporous plate device, preparation method and kit for multi-connection detection of vaginitis

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1729299A (en) * 2002-10-29 2006-02-01 昭和药品化工株式会社 Reagent composition for detecting beta-lactamase, detection kit and detection method
CN101320040A (en) * 2008-07-18 2008-12-10 杭州健宝医疗器械有限公司 Gynaecologic multi-item dry chemical united detection test paper strip and its measuring method
CN101792791A (en) * 2009-12-30 2010-08-04 北京中生金域诊断技术有限公司 Kit for detecting bacteria flora in vaginal discharge and preparation method thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1729299A (en) * 2002-10-29 2006-02-01 昭和药品化工株式会社 Reagent composition for detecting beta-lactamase, detection kit and detection method
CN101320040A (en) * 2008-07-18 2008-12-10 杭州健宝医疗器械有限公司 Gynaecologic multi-item dry chemical united detection test paper strip and its measuring method
CN101792791A (en) * 2009-12-30 2010-08-04 北京中生金域诊断技术有限公司 Kit for detecting bacteria flora in vaginal discharge and preparation method thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
G. CERNY: "Studies on the aminopeptidase test for the distinction of gram-negative from gram-positive bacteria", 《EUROPEAN JOURNAL OF AOOLIED MICROBIOLOGY AND BIOTECHNOLOGY》 *

Cited By (10)

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Publication number Priority date Publication date Assignee Title
CN108642131A (en) * 2018-04-17 2018-10-12 山东仕达思生物产业有限公司 A kind of coagulase detection reagent, reacting pad and preparation method thereof and kit
CN108982493A (en) * 2018-09-17 2018-12-11 迪瑞医疗科技股份有限公司 A kind of neuraminidase drying chemical reagent paper and preparation method thereof
CN109265373A (en) * 2018-11-03 2019-01-25 郑州安图生物工程股份有限公司 A kind of amino acids substrate and its preparation method and application
CN109991221A (en) * 2019-03-29 2019-07-09 迪瑞医疗科技股份有限公司 A kind of preparation method of Prolyl iminopeptidase Test paper
CN109991218A (en) * 2019-03-29 2019-07-09 迪瑞医疗科技股份有限公司 A kind of preparation method of coagulase Test paper
CN111690713A (en) * 2020-06-01 2020-09-22 青岛华晶生物技术有限公司 Eight joint inspection kit of vaginitis
CN113030077A (en) * 2021-03-17 2021-06-25 桂林优利特医疗电子有限公司 N-acetylglucosaminidase detection test paper and preparation method thereof
CN113109333A (en) * 2021-04-30 2021-07-13 桂林优利特医疗电子有限公司 Proline aminopeptidase rapid detection test paper and preparation method thereof
CN114317678A (en) * 2021-12-31 2022-04-12 港龙生物技术（深圳）有限公司 Biological paper chip, high-flux multi-connection detection microporous plate device, preparation method and kit for multi-connection detection of vaginitis
CN114317678B (en) * 2021-12-31 2023-11-10 港龙生物技术（深圳）有限公司 Biological paper chip, high-throughput multi-detection microplate device, preparation method and kit for vaginal inflammation multi-detection

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