CN105039497A - Bacterial vaginosis combined detection reagent and method for using bacterial vaginosis combined detection reagent - Google Patents
Bacterial vaginosis combined detection reagent and method for using bacterial vaginosis combined detection reagent Download PDFInfo
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Abstract
The present invention relates to the technical field of vaginosis detection, and particularly discloses a bacterial vaginosis combined detection reagent and a method for using the bacterial vaginosis combined detection reagent, wherein the bacterial vaginosis combined detection reagent comprises a hydrogen peroxide concentration determination reaction solution, a leukocyte esterase activity determination reaction solution, and a neuraminidase activity determination reaction solution, the reaction solutions are prepared into dried chemical test paper, and whether the bacterial vaginosis exists is determined through the coloration of the dried chemical test paper. According to the present invention, according to the significant change of the activity of the biochemical enzymes in the vaginitis patient vaginal secretions, when the enzyme contacts the color generating substrate, the specific reaction can be generated while the color is changed, the hydrogen peroxide change is combined to prepare the dried chemical test paper, the reaction time only requires 10 min, any equipment is not required, the result can be judged visually and clearly, the immediate inspection requirement can be met, the multi-item combined detection can be simultaneously performed, the accuracy of the diagnosis result is substantially improved, and the influence of cumbersome steps, equipment and subjective factors on the diagnostic result are avoided.
Description
Technical field
The present invention relates to vaginosis detection technique field, the concrete a kind of method disclosing bacterial vaginopathy combined detection reagent and use this reagent.
Background technology
Vaginitis is the inflammation of reticular tissue under vaginal mucosa and mucous membrane, change with the proterties of leukorrhea clinically and pruritus vulvue cusalgia for principal feature, infect when involving urethra, the symptom such as odynuria, urgent urination can be had.Wherein bacterial vaginosis (BV) is modal gynecological inflammation.
At present, diagnose the standard of BV to be clinically, meeting in following 4 indexs 3 diagnosable is BV:1, vagina PH > 4.5; 2, vaginal secretions increases, thinning as milk shape, has peculiar smell; 3, amine test is positive, in vaginal secretions, namely adds 10%KOH produce fishy smell; 4, object line funicular cell in vaginal smear.This is traditional Amsel method.
There is a lot of weak point in the method for conventional diagnostic BV.Major embodiment is, some and infect irrelevant factor as recent sexual intercourse, Vaginal lavages, to pass through or there is the same vagina PH values that can cause such as a large amount of cervical mucuss raises, or make secretion generation peculiar smell; In amine test, the discriminating of smell is varied with each individual; Identification for the clues cell being used as the best mark of BV diagnosis is then subject to the joint effect of the factors such as experience of the quality of microscopy apparatus, the collection of sample and operator, and subjectivity is strong, is thus difficult to add up result of determination and compare.
Summary of the invention
The object of the invention is to provide a kind of bacterial vaginopathy combined detection reagent, and this reagent adopts dry chemistry zymotechnic, by measuring the change of vaginal secretions mesophytization composition, and can effective discriminating bacteria vaginosis.
To achieve these goals, invention have employed following technical scheme: a kind of bacterial vaginopathy combined detection reagent, comprises the sweet enzyme assay reaction solution of concentration of hydrogen peroxide assaying reaction liquid, leukocyte esterase determination of activity reaction solution and sialic acid;
Described concentration of hydrogen peroxide assaying reaction liquid comprises following component and proportioning: 90mlPH value be 6.6 0.1M citrate buffer solution, the sucrose of 1.0g-3.0g, the polyvinylpyrrolidone of 0.5-1.5g, 40mMVC, 0.2g-1.0g of 0.05ml-0.2ml 2, the amino antipyrine of the 750U/ml Peroxidase Solution of the bromo-3-hydroxy-benzoic acid of 4,6-tri-, 5ml-10ml, the 80mM4-of 0.2ml-2ml;
Described leukocyte esterase determination of activity reaction solution is made up of A liquid and B liquid, and component and the proportioning of described A liquid are: the polyvinylpyrrolidone of NaCl, 0.1g-1g of the purified water of 100ml, the sodium tetraborate of 1g-5g, 0.5g-2g, the Sodium cholic acid of 0.1g-0.5g, the magnesium sulfate heptahydrate of 0.5g-2g; Component and the proportioning of described B liquid are: the bromo-4-of the 5-chloro-3-indoles ethyl ester of the white corpuscle stablizer of the anhydrous methanol of 100.0ml, the L-TARTARIC ACID of 0.2g-1.5g, 0.2g-2g, 1,10 decanediols of 0.02g-0.1g, 0.2g-2g;
Component and the proportioning of described sialic acid sweet enzyme assay reaction solution are: the chloro-3-indoles of the bromo-4-of the 5--α-D-N-n acetylneuraminic acid n sodium salt of the polyvinylpyrrolidone of the purified water of 100.0ml, the anhydrous sodium acetate of 0.5g-3g, 0.2g-2g, the sucrose of 0.5g-5g, 0.05g-3g, the 0.02M citric acid of 0.02ml-0.08ml.
Use a method for mentioned reagent,
By concentration of hydrogen peroxide assaying reaction immersion bubble filter paper, dry, make the dry chemistry scraps of paper, namely can be used for detecting; Some chemical film-makings take on a red color or red-purple is feminine gender, and a large amount of lactobacillus that expressed possibility exists, and vaginal microbial flora is normal; Not develop the color or faint yellow for positive, instruction vaginal dysbacteriosis, vaginal environment morbid state or be in sub-health state;
By leukocyte esterase determination of activity reaction solution first with A immersion bubble filter paper, with B immersion bubble after drying, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Blue-greenish colour is positive, shows that patient's intravaginal has inflammatory reaction;
Soak filter paper with the sweet enzyme assay reaction solution of sialic acid, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Aobvious redness or red-purple are positive, and express possibility bacterial infection vaginosis.
The present invention comprises three independent detection projects, is respectively:
1. concentration of hydrogen peroxide measures: in secretory product, hydrogen peroxide is through peroxidase effect, discharges nascent oxygen, and the latter is under the amino antipyrine of 4-exists, make 2, the bromo-3-hydroxy-benzoic acid oxidation of 4,6-tri-takes on a red color or red-purple, is directly proportional to concentration of hydrogen peroxide in color depth.Take on a red color or red-purple for negative, a large amount of lactobacillus that expressed possibility exists, and vaginal microbial flora is normal; Not develop the color or faint yellow for positive, instruction vaginal dysbacteriosis, vaginal environment morbid state or be in sub-health state.
2. leukocyte esterase determination of activity: the leukocyte esterase hydrolysis bromo-4-of 5-chloro-3-indoles ethyl ester, discharge bromo indole base, the latter in blue-greenish colour, is directly proportional to leukocyte esterase activity in color depth under the condition of aerobic.Do not develop the color for feminine gender; Blue-greenish colour is positive, shows that patient's intravaginal has inflammatory reaction.
3. neuraminidase activity measures: the chloro-3-indoles of the neuraminidase hydrolysis bromo-4-of 5--α-D-N-n acetylneuraminic acid n sodium salt, discharge bromo indole base, the latter is aobvious red or red-purple under the condition of diazonium salt.Do not develop the color for feminine gender; Aobvious redness or red-purple are positive, and express possibility bacterial infection vaginosis.
Beneficial effect of the present invention is: after adopting mentioned reagent and method thereof, possess following advantage:
Lactobacillus can produce a large amount of hydrogen peroxide; Clues cell has leukocyte esterase activity; Neuraminidase activity in BV woman vagina secretory product is apparently higher than normal women.
Namely, according to the feature of above-mentioned main biochemical enzymic activity noticeable change in vaginitis patient vaginal secretions, when enzyme contacts with its chromogenic substrate, specific reaction can be there is, and with color change.In conjunction with the change of hydrogen peroxide, it is made drying chemical reagent paper respectively, reaction only needs 10 minutes, and without the need to any plant and instrument, result range estimation judges clear, is very suitable for real-time test (POCT) testing requirement.Entry joint-detection, greatly improves the accuracy of diagnostic result, it also avoid the impact on diagnostic result of traditional diagnosis method complex steps, plant and instrument and artificial subjective factor simultaneously.
Embodiment
Embodiment 1
A kind of bacterial vaginopathy combined detection reagent, concentration of hydrogen peroxide assaying reaction liquid get 90mlPH value be 6.6 0.1M citrate buffer solution, 1.0g sucrose, the polyvinylpyrrolidone of 1.5g, 40mMVC, 1.0g of 0.2ml 2, the amino antipyrine of the 750U/ml Peroxidase Solution of the bromo-3-hydroxy-benzoic acid of 4,6-tri-, 5ml, the 80mM4-of 2ml is made; By concentration of hydrogen peroxide assaying reaction immersion bubble filter paper, dry, make the dry chemistry scraps of paper, namely can be used for detecting; Some chemical film-makings take on a red color or red-purple is feminine gender, and a large amount of lactobacillus that expressed possibility exists, and vaginal microbial flora is normal; Not develop the color or faint yellow for positive, instruction vaginal dysbacteriosis, vaginal environment morbid state or be in sub-health state.
In described leukocyte esterase determination of activity reaction solution, A liquid gets the purified water of 100ml, the sodium tetraborate of 1g, the polyvinylpyrrolidone of NaCl, 0.1g of 0.5g, the Sodium cholic acid of 0.1g, the magnesium sulfate heptahydrate of 2g make; B liquid gets the anhydrous methanol of 100.0ml, the L-TARTARIC ACID of 0.2g, the white corpuscle stablizer of 0.2g, 1,10 decanediols of 0.1g, the bromo-4-of the 5-chloro-3-indoles ethyl ester of 2g are made; By leukocyte esterase determination of activity reaction solution first with A immersion bubble filter paper, with B immersion bubble after drying, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Blue-greenish colour is positive, shows that patient's intravaginal has inflammatory reaction.
Described sialic acid sweet enzyme assay reaction solution gets the purified water of 100.0ml, the anhydrous sodium acetate of 3g, the polyvinylpyrrolidone of 2g, the sucrose of 0.5g, the chloro-3-indoles of the bromo-4-of the 5--α-D-N-n acetylneuraminic acid n sodium salt of 0.05g, the 0.02M citric acid of 0.02ml are made.Soak filter paper with the sweet enzyme assay reaction solution of sialic acid, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Aobvious redness or red-purple are positive, and express possibility bacterial infection vaginosis.
Embodiment 2
A kind of bacterial vaginopathy combined detection reagent, described concentration of hydrogen peroxide assaying reaction liquid get 90mlPH value be 6.6 0.1M citrate buffer solution, the sucrose of 3.0g, the polyvinylpyrrolidone of 0.5,40mMVC, 0.2g of 0.05ml 2, the amino antipyrine of the 750U/ml Peroxidase Solution of the bromo-3-hydroxy-benzoic acid of 4,6-tri-, 10ml, the 80mM4-of 0.2ml is made; By concentration of hydrogen peroxide assaying reaction immersion bubble filter paper, dry, make the dry chemistry scraps of paper, namely can be used for detecting; Some chemical film-makings take on a red color or red-purple is feminine gender, and a large amount of lactobacillus that expressed possibility exists, and vaginal microbial flora is normal; Not develop the color or faint yellow for positive, instruction vaginal dysbacteriosis, vaginal environment morbid state or be in sub-health state.
In described leukocyte esterase determination of activity reaction solution, A liquid gets the purified water of 100ml, the sodium tetraborate of 5g, the polyvinylpyrrolidone of NaCl, 1g of 2g, the Sodium cholic acid of 0.5g, the magnesium sulfate heptahydrate of 0.5g make; Described B liquid gets the anhydrous methanol of 100.0ml, the L-TARTARIC ACID of 1.5g, the white corpuscle stablizer of 2g, 1,10 decanediols of 0.02g, the bromo-4-of the 5-chloro-3-indoles ethyl ester of 0.2g are made; By leukocyte esterase determination of activity reaction solution first with A immersion bubble filter paper, with B immersion bubble after drying, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Blue-greenish colour is positive, shows that patient's intravaginal has inflammatory reaction.
Component and the proportioning of described sialic acid sweet enzyme assay reaction solution are: the chloro-3-indoles of the bromo-4-of the 5--α-D-N-n acetylneuraminic acid n sodium salt of the polyvinylpyrrolidone of the purified water of 100.0ml, the anhydrous sodium acetate of 0.5g, 0.2g, the sucrose of 5g, 3g, the 0.02M citric acid of 0.08ml are made.Soak filter paper with the sweet enzyme assay reaction solution of sialic acid, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Aobvious redness or red-purple are positive, and express possibility bacterial infection vaginosis.
Embodiment 3
A kind of bacterial vaginopathy combined detection reagent, described concentration of hydrogen peroxide assaying reaction liquid get 90mlPH value be 6.6 0.1M citrate buffer solution, the sucrose of 2g, the polyvinylpyrrolidone of 1g, 40mMVC, 0.5g of 0.1ml 2, the amino antipyrine of the 750U/ml Peroxidase Solution of the bromo-3-hydroxy-benzoic acid of 4,6-tri-, 6ml, the 80mM4-of 0.5ml is made; By concentration of hydrogen peroxide assaying reaction immersion bubble filter paper, dry, make the dry chemistry scraps of paper, namely can be used for detecting; Some chemical film-makings take on a red color or red-purple is feminine gender, and a large amount of lactobacillus that expressed possibility exists, and vaginal microbial flora is normal; Not develop the color or faint yellow for positive, instruction vaginal dysbacteriosis, vaginal environment morbid state or be in sub-health state.
In described leukocyte esterase determination of activity reaction solution, A liquid gets the purified water of 100ml, the sodium tetraborate of 2g, the polyvinylpyrrolidone of NaCl, 0.8g of 0.9g, the Sodium cholic acid of 0.2g, the magnesium sulfate heptahydrate of 1.3g make; Described B liquid gets the anhydrous methanol of 100.0ml, the L-TARTARIC ACID of 1g, the white corpuscle stablizer of 1.5g, 1,10 decanediols of 0.04g, the bromo-4-of the 5-chloro-3-indoles ethyl ester of 0.9g are made; By leukocyte esterase determination of activity reaction solution first with A immersion bubble filter paper, with B immersion bubble after drying, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Blue-greenish colour is positive, shows that patient's intravaginal has inflammatory reaction.
Described sialic acid sweet enzyme assay reaction solution gets the purified water of 100.0ml, the anhydrous sodium acetate of 2g, the polyvinylpyrrolidone of 1.5g, the sucrose of 4g, the chloro-3-indoles of the bromo-4-of the 5--α-D-N-n acetylneuraminic acid n sodium salt of 2g, the 0.02M citric acid of 0.05ml are made.Soak filter paper with the sweet enzyme assay reaction solution of sialic acid, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Aobvious redness or red-purple are positive, and express possibility bacterial infection vaginosis.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (2)
1. a bacterial vaginopathy combined detection reagent, is characterized in that, comprises the sweet enzyme assay reaction solution of concentration of hydrogen peroxide assaying reaction liquid, leukocyte esterase determination of activity reaction solution and sialic acid;
Described concentration of hydrogen peroxide assaying reaction liquid comprises following component and proportioning: 90mlPH value be 6.6 0.1M citrate buffer solution, the sucrose of 1.0g-3.0g, the polyvinylpyrrolidone of 0.5-1.5g, 40mMVC, 0.2g-1.0g of 0.05ml-0.2ml 2, the amino antipyrine of the 750U/ml Peroxidase Solution of the bromo-3-hydroxy-benzoic acid of 4,6-tri-, 5ml-10ml, the 80mM4-of 0.2ml-2ml;
Described leukocyte esterase determination of activity reaction solution is made up of A liquid and B liquid, and component and the proportioning of described A liquid are: the polyvinylpyrrolidone of NaCl, 0.1g-1g of the purified water of 100ml, the sodium tetraborate of 1g-5g, 0.5g-2g, the Sodium cholic acid of 0.1g-0.5g, the magnesium sulfate heptahydrate of 0.5g-2g; Component and the proportioning of described B liquid are: the bromo-4-of the 5-chloro-3-indoles ethyl ester of the white corpuscle stablizer of the anhydrous methanol of 100.0ml, the L-TARTARIC ACID of 0.2g-1.5g, 0.2g-2g, 1,10 decanediols of 0.02g-0.1g, 0.2g-2g;
Component and the proportioning of described sialic acid sweet enzyme assay reaction solution are: the chloro-3-indoles of the bromo-4-of the 5--α-D-N-n acetylneuraminic acid n sodium salt of the polyvinylpyrrolidone of the purified water of 100.0ml, the anhydrous sodium acetate of 0.5g-3g, 0.2g-2g, the sucrose of 0.5g-5g, 0.05g-3g, the 0.02M citric acid of 0.02ml-0.08ml.
2. the method using reagent described in claim 1 to detect, is characterized in that: by concentration of hydrogen peroxide assaying reaction immersion bubble filter paper, dry, make the dry chemistry scraps of paper, namely can be used for detecting; Some chemical film-makings take on a red color or red-purple is feminine gender, and a large amount of lactobacillus that expressed possibility exists, and vaginal microbial flora is normal; Not develop the color or faint yellow for positive, instruction vaginal dysbacteriosis, vaginal environment morbid state or be in sub-health state;
By leukocyte esterase determination of activity reaction solution first with A immersion bubble filter paper, with B immersion bubble after drying, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Blue-greenish colour is positive, shows that patient's intravaginal has inflammatory reaction;
Soak filter paper with the sweet enzyme assay reaction solution of sialic acid, then dry, make the dry chemistry scraps of paper, namely can be used for detecting; The dry chemistry scraps of paper do not develop the color for feminine gender, represent that patient's intravaginal is normal; Aobvious redness or red-purple are positive, represent bacterial infection vaginosis.
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| CN108330159A (en) * | 2018-03-15 | 2018-07-27 | 深圳市瀚德标检生物工程有限公司 | A kind of detection reagent card and kit |
| CN109402221A (en) * | 2018-09-28 | 2019-03-01 | 广州江元医疗科技有限公司 | A kind of quick detection kit of bacterial vaginosis BV and its preparation method and application |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106645721A (en) * | 2016-09-30 | 2017-05-10 | 广州鸿琪光学仪器科技有限公司 | Coagulase detection reagent, reaction pad, preparation method thereof and kit |
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| CN108330159A (en) * | 2018-03-15 | 2018-07-27 | 深圳市瀚德标检生物工程有限公司 | A kind of detection reagent card and kit |
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| CN109402221A (en) * | 2018-09-28 | 2019-03-01 | 广州江元医疗科技有限公司 | A kind of quick detection kit of bacterial vaginosis BV and its preparation method and application |
| CN109402221B (en) * | 2018-09-28 | 2019-08-23 | 广州江元医疗科技有限公司 | A kind of quick detection kit of bacterial vaginosis BV and its preparation method and application |
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