Evaluation of the iNtRON VRE vanA/vanB Real-Time PCR Assay for Detection of Vancomycin-Resistant Enterococci.
- Author:
Hee Jae HUH
1
;
Mi Ae JANG
;
Ja Young SEO
;
Ji Youn KIM
;
Chang Seok KI
;
Jong Won KIM
;
Nam Yong LEE
Author Information
- Publication Type:Evaluation Studies ; Original Article ; Research Support, Non-U.S. Gov't
- Keywords: Real-time PCR; Performance; vanA; vanB; Vancomycin-resistant enterococci
- MeSH: Bacterial Proteins/*genetics; Bacterial Typing Techniques/*methods/standards; Carbon-Oxygen Ligases/*genetics; DNA, Bacterial/*metabolism; Gram-Positive Bacterial Infections/microbiology; Humans; Reagent Kits, Diagnostic; *Real-Time Polymerase Chain Reaction; Vancomycin Resistance/genetics; Vancomycin-Resistant Enterococci/*genetics/isolation & purification
- From:Annals of Laboratory Medicine 2015;35(1):76-81
- CountryRepublic of Korea
- Language:English
- Abstract: BACKGROUND: Recently, the iNtRON VRE vanA/vanB real-time PCR (iNtRON; iNtRON Biotechnology, Korea) assay, a multiplex real-time PCR method, was introduced. In this prospective study, we compared the iNtRON assay with the Seeplex VRE ACE detection kit (Seeplex; Seegene, Korea), a conventional multiplex PCR assay. METHODS: A chromogenic agar-based culture, in which pre-selected vancomycin-resistant enterococci (VRE) was grown and subsequently plated on blood agar with vancomycin disks, was regarded as the reference method. A total of 304 consecutive rectal swab specimens were tested for VRE by culture and by iNtRON and Seeplex PCR assays. For the PCR assays, specimens were enriched for 16-24 hr before PCR. RESULTS: VRE were isolated from 44 (14.5%) specimens by chromogenic agar-based culture. The clinical sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the iNtRON assay were 100% (95% confidence interval: 89.8%-100%), 99.2% (96.9%-99.9%), 95.6% (83.6%-99.2%), and 100% (98.2%-100%), respectively, while those of the Seeplex assay were 97.7% (86.2%-99.9%), 99.6% (97.5%-99.9%), 97.7% (86.2%-99.9%), and 99.6% (97.5%-99.9%), respectively. The iNtRON assay had a detection limit of 3,159 copies/microL and 13,702 copies/microL for the vanA and vanB genes, respectively. No cross-reactivity was observed in 11 non-VRE bacterial culture isolates. CONCLUSIONS: The overall performance of the iNtRON assay was comparable to that of a chromogenic agar-based culture method for prompt identification of VRE-colonized patients in hospitals. This assay could be an alternative or supportive method for the effective control of nosocomial VRE infection.
