Evaluation of a Newly Developed Multiplex Real-time PCR Assay for the Detection of Vancomycin-Resistant Enterococci from Rectal Swabs.
10.5145/KJCM.2011.14.4.138
- Author:
Min Kwon JUNG
1
;
Wee Gyo LEE
;
Myung Hwa PARK
Author Information
1. Department of Laboratory Medicine, Seran General Hospital, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Vancomycin resistant enterococci;
Multiplex real-time PCR;
Rectal swab
- MeSH:
Bacteria;
Colon;
DNA;
Enterococcus;
Enterococcus faecium;
Humans;
Limit of Detection;
Mass Screening;
Real-Time Polymerase Chain Reaction;
Sensitivity and Specificity;
Vancomycin
- From:Korean Journal of Clinical Microbiology
2011;14(4):138-143
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Asymptomatic vancomycin-resistant enterococci (VRE) colonization precedes infection. VRE-colonized patients serve as silent reservoirs of enterococci that go on to colonize other patients. Rapidly identifying colonized patients is crucial to prevent the spread of VRE. The culture-based method of VRE screening is time-consuming. We evaluated the diagnostic performance of a recently developed multiplex real-time PCR for the detection of VRE. METHODS: We obtained 105 rectal swabs from patients who were being monitored for carriage of VRE. After 24 hour incubation of swabs in enterococcosel broth (EB) supplemented with 6 microg/mL vancomycin, multiplex real-time PCR was performed using the Anyplex(TM) VanR Real-time Detection (VanR) kit (Seegene, Inc., Seoul, Korea). The results of multiplex real-time PCR were compared to those of culture. We evaluated the specificity and detection limits of multiplex real-time PCR using VanR for VRE. RESULTS: A total of 96/105 (91.4%) samples were VRE positive according to multiplex real-time PCR with EB while 85/105 (80.9%) samples were positive in culture. Eleven discordant results (10.4%) (multiplex real-time PCR positive, culture negative) were noted. All non-enterococcal bacteria and vancomycin-susceptible enterococci were negative. The DNA detection limits of VanR were 0.035 pg per reaction (3 microL) for Enterococcus faecium and 0.35 pg for Enterococcus faecalis. CONCLUSION: The application of multiplex real-time PCR after EB incubation allows rapid and sensitive detection in 26-28 hours for VRE screening from rectal swabs. This method could facilitate the timely implementation of contact isolation to prevent the spread of VRE.
