Work

A multiplex PCR for rapid detection of vancomycin and linezolid resistance genes in clinical and environmental enterococci: A one health diagnostic approach

Year: 2025

Type: article

Abstract: Background Antimicrobial resistance (AMR) in Enterococcus , particularly vancomycin-resistance (VRE) and linezolid-resistance (LZRE) strains, presents a rising threat to public health globally. Methods We designed an assay targeting vanA , vanB , vanC , vanD , optrA , poxtA , cfr and validated it across human and animal Enterococcus isolates. One hundred eighty-six clinical specimens were obtained from patients in a hospital, while 300 environmental isolates were obtained from poultry farms. Analytical sensitivity was assessed using successive dilutions of synthetic DNA constructs and bacterial lysates. The diagnostic performance was benchmarked against the automated system for testing of antimicrobial susceptibility, VITEK II. Results The assay demonstrated high analytical sensitivity, detecting synthetic DNA as low as 1 pg and lysates down to 10 5 CFU/mL. Among 186 clinical isolates, the assay identified VRE genes with a 71.8 % sensitivity, 100 % specificity and an overall 83.6 % accuracy. For LZRE detection, the assay showed 88.9 % sensitivity, 100 % specificity, and 97.1 % accuracy. Statistically significant associations were observed between VRE prevalence and specimen type (χ 2 = 8.77, p = 0.033), with blood samples showing the highest detection rates. In environmental isolates, LZRE genes were detected in 3.0 % of poultry samples. Conclusions This assay supports integrated AMR surveillance within a One Health framework.

Source: One Health

DOI: 10.1016/j.onehlt.2025.101285