I need some information about rapid detection of Carbapenemase enzyme from Gram Negative Bacteria without PCR. Just phenotypic detection techniques are required. Thank you
Hi Fahmidul,
For just phenotypic screening or detection:
Non-automated
1) Disc difusion
2) E-test for MIC determination
Automated antimicrobial susceptibility systems
1) Vitek-2 (Biomerieux, France)
2) Phonenix
3) Microscan walk-Away
4) Micronaut
5) Sensitire etc.
Phenotypic Confirmatory Tests:
1) Modified Hodge test (MHT)
2) EDTA disc test
3) 2-mercaptopropionic acid disc test
4) Boronic acid disc test
Regards
www.ncbi.nlm.nih.gov/pmc/articles/PMC4196366
As an enzymologist, my approach would be to make a soluble extract of a culture of the bacteria and test it for carbapenemase enzyme activity by measuring hydrolysis of a carbapenem using the change in UV absorbance of the compound. This would tell you that a carbapenemase was present and allow for a quantitative assessment of the level of activity, but not which enzyme is responsible.
Thank you everyone. I have already applied carba NP test but some Acinetobacter spp. gave false negative. I don't know why.
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