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First, when to detect based on epidemiological or sensory purposes.
note: Even if the mCIM results are positive, there is no need to modify the results of the carbon penicillin drug sensitivity test.
currently not recommended for routine testing.
II, Method: Merobenan Inactivation Test III, Reagents and Materials 1, TSB (2 ml) 2, Merobenan Sheets (10ug) 3, 1-ul and 10-ul Inoculation Ring 4, Nutritional Broth (e.g. MH Broth, TSB Broth) or MHA plate (100mm or 150mm) 5, Merobenan sensitive quality control strain - E. coli (ATCC25922) IV, step 1, take 1ul inoculation ring overnight culture pure bacteria in 2 ml TSB broth; 2, concussion mix 10-15 seconds; 3, each tube into a sterile piece of paper containing 10ug melo permnam, confirm that the piece of paper immersed in bacterial suspension; 4, 35 degrees C , 2 degrees C atmospheric environment incubation for 4 hours and 15 minutes; 5, at the end of incubation, immediately prepare 0.5 wheat turbidity with nutritious broth or physiological saline e. coli ATCC 25922 bacteria suspension (direct bacteria suspension); 6, bacteria suspension preparation and flat plate coating must be completed within 15 minutes, dry 3-10 minutes; Remove the melopinan pieces from the TSB broth with a 10ul inoculation ring, attach the pieces to the inner wall of the test tube, gently press to squeeze out excess moisture from the sheets, and then remove the pieces from the MHA plate coated with E. coli ATCC25922.
of up to 4 sheets on the MHA tablet with 100mm and up to 8 sheets on the 150mm MHA tablet (Figure 1).
8, inverted plate, 35 degrees C to 2 degrees C atmospheric environment incubation 18-24 hours; 10ul inoculation ring to remove merobean sheet (a); stick the piece of paper to the inner wall of the test tube and squeeze out excess moisture (b); remove the sheet (c) from the same inoculation ring and attach it to MHA plate (d) V, Explanation: See Figures 2a and 2b 1, carbon penicillinase positive: Meropenan antibacterial ring diameter of 6-15mm or diameter of 16-18mm but the bactericidal ring is scattered in the bacterium.
If the strain to be tested produces carbon penicillinase, melopenenan in the paper is degraded by the enzyme, resulting in a reduction in the activity of sterile-free or inhibited E. coli ATCC 25922.
2, carbon penicillinase negative: the diameter of the antibacterial ring is 19mm.
if the strain to be tested does not produce carbon penicillinase, the meropenan in the paper is not hydrolyzed, it can still inhibit the growth of E. coli ATCC 25922.
3, neutral: the diameter of the antibacterial ring is 16-18mm, it is not possible to determine whether there is carbon penicillinase (see note) Figure 2a. Negative control BAA1706 (A) and positive control BAA1705 (B) Figure 2b. Carbon penicillinase production strain.
Note: mCIM results positive Six, Notes: 1, neutral results a) check the purity of strains to be tested and E. coli ATCC 25922 b) Check melopnan quality control results c) repeat mCIM test d) if the results Still neutral, it is recommended to test carbon penicillinase gene 2, some strains may appear scattered in the meropenine anti-bacterial circle, if the diameter of the anti-bacterial ring is 18mm, the result is positive;
3, CLSI currently only recommends this standard method for the detection of carbon penicillin enzymes in E. coli bacteria Seven, report: 1, positive: report "detected carbon penicillinase" 2, negative: report "no detection of carbon penicillin" enease " 3, neutral: report "cannot determine whether carbon penicillinase is produced, contact the laboratory for discussion" Eight, additional tests and reports 1, reporting mCIM results for infection control or requesting epidemiological information 2, even if the mCIM results are positive, There is no need to modify the sensitivity test results of carbon penicillin drugs 9, quality control: 1, daily test positive and negative nature control strain 2, carbon penicillinase positive strain: pneumonia Kreb bacteria ATCC BAA1705 3, carbon penicillin Enzyme-negative strains: Pneumonia Kreber ATCC BAA1706 4, meropenan sheet quality control according to CLSI requirements 5, Source: CLSI M100S-27th Source: Microbiology House.