Analysis of Clinical Isolates of Extended-Spectrum β-Lactamase-Producing Bacteria with Primer and Probe Sets Developed to Detect blaCTX-M, blaTEM, and blaSHV Using a Fully Automated Gene Detection System

Abstract

In this study, we evaluated extended-spectrum β-lactamase (ESBL)-producing bacteria with the newly developed primer and probe sets to detect bla_CTX-M, bla_TEM, and bla_SHV using BD MAX^TM, a fully automated multiplex polymerase chain reaction assay system. In 36 isolates confirmed by whole-genome sequencing to have bla_CTX-M, bla_TEM, or bla_SHV, the developed primer and probe sets accurately detected each gene without being influenced by the presence of other β-lactamase genes. In nine control strains that do not harbor either bla_CTX-M, bla_TEM, or bla_SHV no cross-reaction was observed. In 191 strains phenotypically determined to be ESBL-producers by conventional antimicrobial susceptibility tests, 189 strains were blaC_TX-M-, bla_TEM-, or bla_SHV-positive as assessed by BD MAX^TM using the developed primer and probe sets, and two strains were negative for these genes. Whole-genome sequencing revealed that these two strains were phenotypically false-positive ESBL-producers. The accuracy of the primer and probe sets seems to be satisfactory, and they may be applicable to detect CTX-M-type ESBL-producing bacteria.

Keywords: ESBL; blaCTX-M; blaSHV; blaTEM; fully automated PCR.