Distribution of Class B and Class A β-Lactamases in Clinical Strains of : Comparison of Phenotypic Methods and High-Resolution Melting Analysis (HRMA) Assay

Overview

Journal Infect Drug Resist

Publisher Dove Medical Press

Date 2020 Jul 9

PMID 32636657

Citations 9

Authors

Sanaz Dehbashi

Hamed Tahmasebi

Mohammad Yousef Alikhani

Fariba Keramat

Mohammad Reza Arabestani

Affiliations

Soon will be listed here.

Abstract

Background: There are various phenotypic methods for identifying class B and class A β-lactamase enzymes in . The purpose of this study was to compare the sensitivity and specificity of different phenotypic methods with HRMA assay to detect β-lactamase-producing strains.

Methods: Eighty-eight of isolates were collected from different specimens. Conventional double-disk test (DDT) and EDTA-imipenem microbiological (EIM) were performed to detect ESBL and MBL-producing strains, respectively. Meanwhile, the Modified Hodge test and Carba-NP test were performed on all carbapenem-resistant strains. HRMA method and sensitivity and specificity of primers were determined based on the melt curve temperature range. In all comparisons, PCR was considered as the gold standard.

Results: Of the 402 isolates collected from different clinical specimens, 88 isolates of were identified. However, 43 strains were (48.88%) ESBL-producing, and 7 strains (7.95%) were MBL-producing. Also, using the Modified Hodge test and Carba-NP method, 11 (12.5%) and 19 (21.59%) strains were carbapenemase-producing, respectively. The results of the HRMA test revealed that genes coding for , , , , and were detected in 44.31%, 22.72%, 13.63%, 14.7%, 5.6%, and 2.27% of isolates. Nonetheless, for and genes, sensitivity and specificity of the Carba-NP test were 90.47%, 94.87%, and 83.36%, 94.80%, respectively. However, sensitivity and specificity of MHT was 91.66%, 98.70%, and 77.77%, 96.42%, respectively. For and genes, sensitivity and specificity of DDT were 95.55%, 95.55%, and 86%, 83.50%, respectively. However, sensitivity and specificity of EMI were 77.77%, 97.59%, and 91.66%, 97.43% for and , respectively.

Conclusion: The HRMA is a powerful, accurate, closed-tube, rapid method for detecting β-lactamase genes in . The high sensitivity and specificity of this method, along with phenotypic tests, play a useful role in increasing the predictive value of clinical reports.

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