Microbial (fungus) identification and drug susceptibility analysis system test plate (colorimetric/turbidimetric method)

User Manual (V1.0)

【Product name】 Common name: Microbial (fungus) identification and drug susceptibility analysis system test board (colorimetric/turbidimetric method)

[Packaging Specifications] 10 servings/box.

【Intended use】 It is used for the identification of fungi and semi-quantitative analysis of MIC of antibacterial drugs.

【Inspection Principle】

The system test plate is composed of biochemical reaction wells and antibacterial drug MIC determination test wells. Fungal assimilation culture solution is added to the biochemical reaction wells, and bacterial drug susceptibility culture solution is added to the antibacterial drug MIC determination test wells, and incubated at 35~37°C. The biochemical reaction and antimicrobial drug MIC determination test determine whether there is bacterial growth according to whether the test hole appears turbid (precipitation), and through microbial identification and drug susceptibility analysis system analysis, the bacteria can be quickly and accurately identified to the species and the antimicrobial drug MIC value is analyzed at the same time.

【Main components】 Kit composition:

  1. Fungal identification and drug susceptibility analysis system test board 10 pieces

  2. Diluent 10 pieces

3. Assimilation culture medium 10 pieces

4. Drug susceptibility culture medium 10 sticks

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

【Applicable instruments】 Microbial identification drug sensitivity analysis system.

【Sample requirements】 The tested bacteria using this test board should meet the following requirements:

1. Freshly isolated single pure colony (or pure culture);

2. The culture was confirmed to be a yeast-like fungus by microscopic examination.

【Testing method】

1. Preparation of bacterial solution: Pick a single colony of pure culture and grind it into a bacterial suspension on the inner wall of the diluent bottle, so that the turbidity is 1 McFarland unit.

2. Use a continuous pipette (with no tip) to aspirate 100 μl of the drug-sensitive medium and add it to well E12.

3. Use a continuous pipette to pipette 100 μl of bacterial suspension into the assimilation medium, mix well, and add 100 μl to wells C1-C12 and D1-D12 of the test plate.

(Or use quick sample addition: aspirate 100 μl of bacterial suspension, add it to the assimilation medium and mix well, pour it into the groove on the right side of the test plate, install 6 tips with an 8-channel pipette, and add it to the test plate in 4 times. In wells C1-C12, D1-D12, 100μl per well.)

4. Pipette 50 μl of bacterial suspension into the drug-susceptible culture medium, mix well, and add to the remaining drug-susceptible wells and C+ wells, 100 μl per well.

(Quick sample addition: Aspirate 50μl of the bacterial suspension and add it to the drug-susceptibility culture medium, mix well, pour it into the groove on the left side of the test plate, install 6 tips with an 8-channel pipette, and add the drug to the test plate in 11 times. Sensitive well and C+ well, 100 μl per well.)

5. Tear off the self-adhesive (align the holes) and stick it on the identification plate (the grooves on both sides of the identification plate are also attached), and incubate in a humidified box at 30°C.

6. Take 2 drops of bacterial suspension into a small test tube, add 0.5ml of human/sheep/calf serum, incubate at 35°C for 3 hours, and immediately check the formation of germ tubes with a microscope.

7. Incubate the test plate for 24-72 hours and observe it daily. When the C+ positive control well becomes obviously turbid, immediately interpret the result.

【Explanation of test results】

Analyze the reagent plate with the microbial identification and drug susceptibility analysis system. After the analysis is completed, the bacterial species and MIC semi-quantitative results are automatically determined and printed.

Print the report sheet (the detailed operation of the microbial identification drug susceptibility analysis system is introduced separately).

Table A  Antibacterial drug table of drug susceptibility test

【Limitations of the test method】  

The identification drug susceptibility analysis system is to identify bacteria according to the background data of the database. Incomplete database data will directly affect the accuracy of the identification.

【Product performance index】  

1. Accuracy rate: The accuracy rate of the system test board for the identification of quality control strains should be 100%; the accuracy rate of the system test board for the determination of MIC of quality control strains should be 100%.

2. Repeatability: The repeatability of the system test board for the identification of quality control strains should be 100%; the repeatability of the system test board for the MIC determination of quality control strains should be 100%.

3. Intra-assay imprecision: There should be no difference in the intra-assay imprecision of the system test plate for the identification of quality control strains; there should be no difference in the intra-assay imprecision of the system test plate for the MIC determination of quality control strains.

4. Inter-batch imprecision: There should be no difference in the batch-to-batch imprecision of the system test plate for the identification of quality control strains; there should be no difference in the batch-to-batch imprecision of the system test plate for the MIC determination of the quality control strains.

【Precautions】

1. Compare the assimilation test with the positive control well. If the turbidity of the assimilation test well is higher than that of the blank control well, it is judged as positive; if the turbidity of the assimilation test well and the blank control well is consistent, it is judged as negative.

2. The turbidity of the drug susceptibility test is positive, and the clearness is negative.

3. For fluconazole FLZ, ketoconazole KET, flucytosine FLU, and voriconazole VRC, due to residual growth, the drug susceptibility test should be

Compared with the long control well, the MIC value is determined as the lowest drug concentration at which the growth is reduced by 50% or more relative to the positive growth control.

4. The germ tube test needs to take another sterile small test tube for operation. This test is the key test to identify Candida albicans. The operation and microscopic examination should be strictly in accordance with the regulations.

Many animal serums (rabbit, sheep, cattle, human, etc.) and egg whites can be used, calf serum and sheep serum are better. Be sure to incubate for about three hours for microscopic examination. When observing, take a drop of the incubated serum and place it on a glass slide. Immediately examine it with a low-power microscope. The germ tube looks like an appendage of a yeast cell, about 1/2 the width of the cell body, and the length is about 1/2. About 3 to 4 times that of the cell body, the junction between the germ tube and the cell body does not shrink, and the initial mycelium produced by some yeast-like bacteria cannot be mistaken for the germ tube.

       The difference between germ tube and starting mycelium is shown in the table below

5. The tested bacteria must be fresh and pure cultures, strictly avoid using two or more colonies or using old cultures for testing.

6. The substrates in each well of this test plate are freeze-dried under sterile conditions, sealed and packaged, and then pre-warmed at room temperature before being opened in a sterile environment.

The plate film should not be opened at will to prevent the contamination of air bacteria, and all the operation process should avoid the contamination of bacteria as much as possible.

7. Necessary protective measures must be taken during the operation to prevent the operator from being infected, and the waste should be treated as a source of infection.  

8. This product is only used for in vitro diagnosis and is for one-time use. It is forbidden to leak or damage the package of the reagent plate. Diluent and medium bottle

Abnormal phenomena such as turbidity and precipitation appear in the product and should not be used.  

9. Please read this manual carefully before use.

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

3. "National Clinical Inspection Operating Regulations"

 4.《Clinical and Laboratory Standards Institute》document

【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