CN1401789A

CN1401789A - Paper disk method amine test detection of gardneri and method for producing amine test paper

Info

Publication number: CN1401789A
Application number: CN 01128396
Authority: CN
Inventors: 宋绍业
Original assignee: Individual
Current assignee: Individual
Priority date: 2001-08-28
Filing date: 2001-08-28
Publication date: 2003-03-12
Anticipated expiration: 2021-08-28
Also published as: CN1229503C

Abstract

A process for preparing the amine test paper for detecting the infection of vigna with Gardner's bacteria includes dissolving the bromocresol green and bromocresol purple in alcohol, diluting with distilled water, soaking ordinary filter paper in it, and drying. Its application includes such steps as sampling the vigna secretion, adding alkali solution and putting said test paper in it. If the colour of test paper changes, said infection can be determined. Its advantages are high speed and high correctness.

Description

The paper disk method amine test detects the preparation method of Gardnerella and amine test paper

Technical field

The present invention relates to the detection method for preparing test paper of test paper, particularly Gardnerella that a kind of woman vagina germ detects and the detection method of Gardnerella.

The important germ of background technology gardnerella vaginalis (Gardnerolla Vaginalis.av) the special vaginosis of right and wrong, confirmed that this bacterium can be through transmission through sex, cause illnesss such as septicemia, urinary tract infections, perinephric abscess or vagina abscess, and can cause multiple complications such as premature labor, chorion sheep inflammation, salpingitis, endometritis, uterus tumor etc., so the Obstetric and Gynecologic Department of hospital and STD Dept., department of infectious disease are usually detected Gardnerella, so that symptomatic treatment.Because it is harsh that the Gardnerella growth requires, be difficult to obtain pure culture, thereby the detection method of this bacterium all is in always explores and conceptual phase, so medical journal has been delivered the method for some check Gardnerellas in recent years, having introduced Gardnerella in " Harbin Medical University's journal " the 1st phase in 1998 " gardnerella vaginalis " literary composition has several methods of inspection: one, microscopy; Two, amine test method; Three, secretory product PH assay method; Four, fluorescent antibody technique; Five, vapor-phase chromatography; Six, bacterium autoanalyzer method; Seven, polymerase chain reaction,PCR method (PCR method); The method of inspection that " Clinical Laboratory magazine " the 4th phase in 1992 discloses Gardnerella is to be substratum with multivalence peptone, Tryptones, beef extract, yeast extract, amino acid, salt, glucose, the adding ascites pleural fluid is a nutritional additive, cultivate the back smear earlier, with gram staining method and acridine orange dyeing, microscopy; " National Medical College Of Right Rivers's journal " 1999 the 21st volumes propose to detect gardnerella vaginalis with polymerase chain reaction (PCR) method, and required sample, reagent, instrument are all available from abroad.

Higher, the troublesome poeration of the most cost of the detection method of above-mentioned disclosed several Gardnerellas needs special plant and instrument, and the policlinic of only having ready conditions could be implemented, and many general hospitals are difficult to purchase top-grade instrument, thereby can't carry out testing.

Technology contents

In order to overcome the deficiency of prior art, the inventor is through in a few years arduous exploration and research, found a kind of senior plant and instrument that do not need, detection method is simple, be fit to large hospital, the infection of middle and small hospital outpatient service routine inspection Gardnerella, also be fit to health clinics in towns and townships's check vaginal infection Gardnerella diagnosis.The preparation method of the amine test paper of paper disk method amine test is as follows:

1, tetrabromo-mcresolsulfonphthalein and purpurum bromocresolis are dissolved in dehydrated alcohol earlier by 2: 3 weight ratio, are made into 1--2% stoste.

2, stoste is diluted with distilled water or deionized water, making its concentration is between the 0.05%-0.5%.

3, with common filter paper with distilled water or rinsed with deionized water 2-3 time, take out dry, dye from the lower concentration staining fluid, till dying to 0.5%, after dying at every turn, all will be 60-80 ℃ temperature oven dry, until obtaining faint yellow or brown xanchromatic test paper, decide used specification, be encapsulated in then in plastics bag or the drying receptacle and preserve.This test paper did not lose efficacy in room temperature in 3 years.

The detection method of Gardnerella

1, material and reagent: amine test paper, 2.0mol/l NH4Cl standardized solution;

2, operation: get 3 penicillin bottles, perhaps akin vial is indicated assay flask, positive control bottle, negative control bottle.Assay flask, every in positive control bottle add 0.5ml, and the negative control bottle adds 0.6ml does not have ammonia physiological saline.Assay flask adds tested vaginal secretions, and positive bottle adds the 0.1mlNH4Cl standardized solution, and each bottle adds 1 of 10%KOH solution then, shakes up, and covers at bottleneck with amine test paper a slice, pushes down with slide, sees the result, test temperature 25-35 ℃ in 10 minutes.It is positive that circular even bluish voilet appears in the amine test paper on bottleneck, and nondiscoloration is negative.When the assay flask test paper becomes bluish voilet, the report paper disk method amine test positive, nondiscoloration is reported as feminine gender.

Compare with current domestic and international similar technology:

1, smell method amine test method for method one of is, is to be proposed by pheifer in 1978, in secretory product, add 10%KOH solution, hear it with nose near secretory product and whether send " raw meat sample " ammonia stink, differentiate negative by sense of smell, subjective factor is big, and the result is unreliable.

2, line inspection funicular cell diagnosis is method for it two, and this method takes time longer, and peopleware requires high, and accuracy is also not too high, can not use as routine.

3, use the cultural method diagnosis, be three of method, this method is " gold standard ", in case cultivate to come out can make a definite diagnosis, but this method is more time-consuming, goes out report in 3-5 days, and equipment is many, conditional request height, peopleware require higher, usually County Hospital all extremely difficulty carry out this cultivation.

Characteristics of the present invention are, utilize the amine material that has Gardnerella in the vaginal secretions to meet the principle that alkali decomposites ammonia, make the variable color of amine test paper, this method fast, simple, accurately, make things convenient for outpatient service to do routine inspection and Quality Control.Test paper good stability, sealing are preserved and were not lost efficacy in 3 years.

Embodiment

Embodiment one: take by weighing 20 gram tetrabromo-mcresolsulfonphthaleins and 30 gram purpurum bromocresoliss, earlier with 50 milliliters of anhydrous alcohol solutions, use 1 liter of deionized water dissolving then, stir, get common filter paper and soak into the solution that obtains, take out dry, soak, drying repeats 3 times again, 60-90 ℃ oven for drying, be cut into 2.5*2.5cm then ²The scraps of paper are preserved in plastics bag or drying receptacle.

Embodiment two, take by weighing 50 gram tetrabromo-mcresolsulfonphthaleins and 80 gram purpurum bromocresoliss with 0.2 liter of anhydrous alcohol solution, add 3 liters of dissolved in distilled water then, stir, get common filter paper and be immersed in this solution, take out dryly, soak again, drying, repeat 4 times, 60-90 ℃ oven for drying, be cut into 5*5cm then ²The scraps of paper are preserved in drying receptacle or plastics bag, cut out little before the use again.

The test experience data of Gardnerella

1, the inventor contrasts with the inventive method and the blue color method of leather, get 573 parts of vaginal secretions smears of the out-patient of provincial hospital gynaecology at random, bacterium 〉=11 of gram's staining microscopy gardnerella vaginalis form bacterium and genera bacillus form/HP person accounts for 74 parts, 499 parts of normal vaginal secretionss, positive 71 parts of present method, negative 502 parts.X2 check P＞0.05, two group difference nonsignificance, its coincidence rate 99.5%, susceptibility 95.9%, specificity 100%.

2,294 parts of secretory product are checked in present method and former amine test method contrast, negative 21 parts of former amine test, and (comprising 9 parts of probable positives), positive 36 parts of the inventive method test is handled through X2, and P＜0.01 shows that two kinds of test method differences have the highly significant meaning.

3, the inventive method and line inspection funicular cell method contrast: 1368 parts of vaginal secretions samples of gynaecology's outpatient service are done the inventive method test, positive 170 parts, with 170 parts of positive patient secretory product line inspection funicular cells, 149 parts of positives are arranged, both coincidence rates 87.6%.

4, the present invention and cultivation Gardnerella contrast: test 457 parts of vaginal secretionss altogether, 44 strain Gardnerellas are isolated in positive 37 parts of the inventive method test, and susceptibility 84.1% by statistics, specificity 100%, positive prediction 100%, negative predictive value 98.3%.

Claims

1, a kind of detection method for preparing test paper of Gardnerella is characterized in that:

(1) tetrabromo-mcresolsulfonphthalein and purpurum bromocresolis are dissolved in dehydrated alcohol earlier by 2: 3 amount and are made into 1-2% stoste;

(2) stoste is diluted with distilled water or deionized water, making its concentration is between the 0.05%-0.5%;

(3) with common filter paper with distilled water or rinsed with deionized water 2-3 time, take out dry, dye from the lower concentration staining fluid, till dying to 0.5%, after dying, all in 60-80 ℃, dry, until obtaining faint yellow or brown xanchromatic test paper at every turn, decide used specification, be encapsulated in then in plastics bag or the drying receptacle and preserve.

2, a kind of method that adopts the described detection paper Gardnerella of claim 1, it is characterized in that: get 3 penicillin bottles, perhaps akin vial, indicate assay flask, the positive control bottle, the negative control bottle, at assay flask, every adds 0.5ml in the positive control bottle, the negative control bottle adds 0.6ml does not have ammonia physiological saline, assay flask adds tested vaginal secretions, positive bottle adds the 0.1mlNH4Cl standardized solution, and each bottle adds 1 of 10%KOH solution then, shakes up, cover at bottleneck with amine test paper a slice, push down with slide, saw the result, test temperature 25-35 ℃ in 10 minutes.It is positive that circular even bluish voilet appears in the amine test paper on bottleneck, and nondiscoloration is negative.