1887

Abstract

Carbapenem-resistant pathogens cause infections associated with significant morbidity and mortality. This study evaluates the use of the loop-mediated isothermal amplification (LAMP) assay for rapid and cost-effective detection of and genes among carbapenem-resistant Gram-negative bacteria in comparison with conventional PCR and existing phenotypic methods. A total of 60 carbapenem-resistant clinical isolates [ (15), (22), (23)] were screened for the presence of carbapenemases ( and ) using phenotypic methods such as the modified Hodge test (MHT) and combined disc test (CDT) and molecular methods such as conventional PCR and LAMP assay. In all, 47/60 isolates (78.3 %) were MHT positive while 48 isolates were positive by CDT [46.6 % positive with EDTA, 30 % with 3′ aminophenylboronic acid (APB) plus EDTA and 1.6 % with APB alone]. Isolates showing CDT positivity with EDTA or APB contained and genes, respectively. was present as a lone gene in 28 isolates (46.7 %) and present together with the gene in 19 isolates (31.7 %). Only one isolate had a lone gene. The LAMP assay detected either or both and genes in four isolates that were missed by conventional PCR. Neither gene could be detected in 12 (20 %) isolates. The LAMP assay has greater sensitivity, specificity and rapidity compared to the phenotypic methods and PCR for the detection of and . With a turnaround time of only 2–3 h, the LAMP assay can be considered a point-of-care assay.

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2013-10-01
2020-01-25
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