Mycoplasma Identification Drug Susceptibility Kit (Microbial Culture Method)

Instruction Manual (V1.0)

【Product name】Generic name: Mycoplasma identification drug susceptibility kit (microbial culture method)

[Packaging Specifications] 20 servings/box

【Intended use】Suitable for the isolation and identification of Ureaplasma urealyticum and Mycoplasma hominis and drug sensitivity test.

【Testing principle】 This kit is suitable for the separation and identification of Ureaplasma urealyticum and Mycoplasma hominis and drug sensitivity test. It consists of mycoplasma culture medium and test card. The medium contains mycoplasma basal broth, horse serum, yeast extract, phenol red indicator, mixed antibiotics, growth factors and other substances, and the diluent contains urea and arginine. When Uu and Mh grow, urea and arginine The alkaline substance generated by acid decomposition causes the pH value to rise, and the medium changes from yellow to red. Bacteriostatic agents are added to the medium to inhibit the growth of bacteria and fungi in the reproductive tract. The test card medium is produced by freeze-vacuum drying process, which improves the sensitivity, specificity and stability of the reagent.

【Main components】

1. 20 bottles of dry powder medium

2. 20 test cards

3. Sterile mineral oil 12ml * 2 bottles

4. 20 bottles of diluent

5. 1 copy of the instruction manual

The main components of the medium: basal powder 25.5, horse serum 200 ml, yeast powder 4g, 4g/L phenol red 4ml, mixed antibiotic 1.5ml, urea 5g and arginine 5g.

Test card: It consists of upper and lower rows. They are: C-blank wells, C+ control wells, UU identification wells, UU counting wells, Mh identification wells, Mh counting wells, and the remaining 26 wells are drug test wells. For each drug, there are two upper and lower holes, the upper hole is high concentration, the lower hole is low concentration, the drugs are tetracycline (TET concentration 8/4mg/L), erythromycin (ERY concentration 4/1mg/L), doxycycline (DOX concentration 8/4mg/L), Ofloxacin (OFL concentration 4/1mg/L), Josamycin (JOS concentration 8/2mg/L), Azithromycin (AZI concentration 4/0.12mg/L), Clarithromycin (CLA 4/1mg/L), Ciprofloxacin (CIP 2/1mg/L), Levofloxacin (LEV 4/1mg/L), Roxithromycin (ROX 4/1mg/L) ), sabafloxacin (SPA concentration 4/1mg/L), minocycline (MIN concentration 8/4mg/L), dirithromycin (DTM concentration 4/1mg/L).

【Storage conditions and validity period】 Store at 2～8℃, valid for 12 months.

【Sample requirements】

Specimens must be taken before medication. Mycoplasma has a strong affinity for the cell surface. As many cells as possible should be collected. Specimens should be inoculated as soon as possible after collection. die.

Male: Use male sterile swab to take urethral secretions from 1-2 cm from the anterior urethra. According to the needs of the disease, prostate massage fluid and semen can also be taken.

Women: When taking cervical specimens, the excess mucus should be wiped off the cervix first, and the cervical secretions should be taken from the 1-2 cm of the cervical canal with a female swab, and then another swab should be used to obtain the material. When taking the swab, rotate and hold in the cervix (or urethra) for at least 20 seconds to obtain more cells.

If urine must be used for mycoplasma culture, 10-20ml of urine in the middle of the morning can be collected. After high-speed centrifugation, the sediment is inoculated. However, the positive rate of urine mycoplasma culture is significantly lower than that of urethral (or cervical) secretions.

【Testing method】

This test should pay attention to aseptic operation to avoid contamination.

1. Add the diluent to the dry powder medium, mix well, suck up 100 μl of the medium with a sterile pipette and add it to the C-blank hole of the detection card.  

2. Insert the collected specimen swab into the culture bottle, squeeze and rotate the swab several times on the bottle wall near the liquid surface, so that the sample in the swab penetrates; if it is a semen or prostate fluid sample, add 200 μl to the culture medium; If the urine is midstream, centrifuge at 2000 rpm for 10 minutes, and add 100 μl of the sediment to the culture medium.

3. Thoroughly mix the culture medium of the inoculated specimen, and add 100 μl to each well of the test card (except C-well).

4. Add 2 drops of sterile mineral oil dropwise to each well, cover the detection card cover, place it in an incubator at 35°C to 37°C, and observe the results at 24h and 48h respectively.

5. Fill in the manual interpretation results in the report form, or enter the mycoplasma analysis system software (introduced separately) to automatically print the report.

【Explanation of test results】

1. Determination criteria for negative and positive results: yellow or orange → negative (-); clear and transparent red → positive (+); cloudy red → pollution.

2. C-well (blank well) should be negative. If it is positive, it means there is contamination, and this test is invalid.

3. The C+ well (control well) is negative (48h), which can report no Uu and Mh growth.

4. If the C+ well (control well) is positive, the results of the remaining wells should be observed and recorded, and interpreted as follows:

【Limitations of the test method】

1. The test results largely depend on the collection of specimens, so a negative result does not confirm that there is no infection.

2. A positive result indicates the presence of Mycoplasma in the urogenital tract, but it cannot be used as a sufficient basis for clinical diagnosis. The clinical diagnosis should be combined with clinical symptoms.

【Product performance index】

1. Culture medium: lyophilized powder in the form of yellow powder, the diluent is clear liquid, no turbidity, precipitation, and clot, pH is 6.0±0.5 (25℃),

2. Conformity rate: The conformity rate of the kit for the identification of Ureaplasma urealyticum standard strain ATCC27813 and Mycoplasma hominis standard strain ATCC14027 is 100%, and the conformity rate of drug susceptibility is 100%.

3. Repeatability: The repeatability of the kit for Ureaplasma urealyticum standard strain ATCC27813 and Mycoplasma hominis standard strain ATCC14027 is 100%, and the repeatability of drug susceptibility is 100%.

4. There should be no difference in intra-assay and inter-assay imprecision.

【Precautions】

1. Please pay attention to the validity period, use within the validity period, and do not mix reagents of different batch numbers.

2. Menstrual period, women with threatened abortion are prohibited, and it is prohibited after vaginal medication or lavage.

3. Read the instructions carefully before use, and should not use if the dilution is found to be turbid or precipitated before use.

4. Specimens should be collected correctly, and timely inoculation of culture medium and operating procedures are the keys to improving the positive detection rate and accuracy.

5. After the medium is reconstituted, it must be shaken well before inoculating the test card. If the concentration of the sample is very low, the color of the micropores of the test card may not change or the color changes unevenly.

6. The samples were tested for drug susceptibility, which did not consider the concentration of mycoplasma in the sample. In the experiment, it may be observed that the actual sensitivity of the strain caused by the difference in inoculation concentration is different from the results measured by the test card.

7. The growth medium of mycoplasma should be kept clear and clear. The turbidity and redness caused by the contamination of bacteria should not be reported as positive for mycoplasma (but some samples with more secretions may be inoculated into the medium, which may cause the clarity of the medium to decrease. Please note that the difference).

8. Because most of the specimens are inhomogeneous secretions, or the concentration of mycoplasma in the specimens is low, the inoculation amount of the microwells of the test card may be uneven, and the results of the drug sensitivity test are unreasonable (high concentration holes have mycoplasma growth, low concentration holes have mycoplasma growth, low concentration holes no mycoplasma growth). Such experimental results are meaningless, please redo the drug sensitivity test or discard this drug.

9. This product is only used for in vitro diagnosis and is for one-time use. If the outer packaging of the test card is damaged, please do not use it.

10. The operation process must have necessary protective measures to prevent the operator from infection. Waste should be treated as a source of infection.

11. Do not use if the outer packaging is damaged.

【Explanation of the logo】

 ：In vitro diagnostic medical devices.

  ：Refer to the instructions for use.

：Not to be used twice

【references】

1.  Patrick R.Murray, Mannal of clinical Microbiology, 7th edition.

2.  Glatt,A.E.,W.M.Mc Cormack. and D. Taylor-Robison 1989. Genital mycoplasma , P279-293 in sexually Transmitted Disease,2nd , McGraw-Hill Book co., New York, N.Y.

3. Wen Yumei, Modern Medical Microbiology, First Edition, Shanghai Medical University Press.

4. Dai Ziying, Practical Antimicrobials, Second Edition, Shanghai Science and Technology Press.

5. Compiled by Wang Xiancai, Contemporary STD Diagnosis and Treatment, First Edition, Qingdao Publishing House.

6. Ye Shunzhang, Laboratory Diagnosis of Sexually Transmitted Diseases, 1st Edition, Science Press.

【Basic Information】

Name of the registrant/manufacturer: Zhuhai Meihua Medical Technology Co., Ltd.

Address: Unit 3B-3, 3rd Floor, Annex Building, Building 10, Science and Technology 1st Road, Gangwan Avenue, Tangjiawan Town, Zhuhai City

Contact: (0756) 3865699

Postcode: 519085

Tel: (0756) 3865688 Fax: (0756) 3865669

http://www.meihuayl.com 

Name of after-sales service unit: Zhuhai Meihua Medical Technology Co., Ltd.

Contact: 400-603-1358

Production address: Unit 3B-3, 3rd Floor, Annex Building, Building 10, Science and Technology 1st Road, Gangwan Avenue, Tangjiawan Town, Zhuhai City

Production license number: Guangdong Food and Drug Administration Production Xu No. 20081652

【Medical device registration certificate number/product technical number】

Medical device registration certificate number:

Product technical requirement number:

【Approval date and revision date of the specification】 June 1, 2015