DEVELOPMENT OF A NOVEL MULTIPLEX PCR ASSAY FOR THE DETECTION OF MOBILE CARBAPENEM, TETRACYCLINE RESISTANCE GENES, AND CLASS 1 INTEGRON IN ENVIRONMENTAL AND CLINICAL SPECIMENS

Tóm tắt

The rapid spread of antibiotic-resistant bacteria and antimicrobial resistance genes threatens the global health. Polymerase chain reaction based assays offer rapid detection of antibiotic-resistant bacteria and resistance determinants. This study develops a novel multiplex polymerase chain reaction assay for the simultaneous detection of genes: tetM (tetracycline resistance), blaNDM and blaKPC (carbapenem resistance), and intI1 (class 1 integron-integrase gene). The assay demonstrated 100% sensitivity and specificity for the detection of these target genes, both individually and in mixtures. The assay’s limit of detection was established at 10¹ copies/mL for blaKPC and blaNDM, and at 10⁴ copies/mL for tetM and intI1. Furthermore, the multiplex polymerase chain reaction assay was successfully applied to the direct screening of target genes in environmental and clinical samples. In conclusion, the multiplex polymerase chain reaction assay developed in this study represents a valuable tool for the surveillance of tetM, blaNDM, blaKPC, and intI1 genes, and for the detection of antibiotic-resistant bacteria.