Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA.
10.3343/alm.2017.37.6.494
- Author:
Hee Jae HUH
1
;
Ji Youn KIM
;
Hyeon Jeong KWON
;
Sun Ae YUN
;
Myoung Keun LEE
;
Chang Seok KI
;
Nam Yong LEE
;
Jong Won KIM
Author Information
1. Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. changski@skku.edu
- Publication Type:Original Article
- Keywords:
MERS-CoV;
Real-time reverse-transcription PCR;
Performance
- MeSH:
Coronavirus Infections*;
Diagnosis;
In Vitro Techniques;
Limit of Detection;
Methods;
Middle East Respiratory Syndrome Coronavirus*;
Middle East*;
Real-Time Polymerase Chain Reaction*;
Reverse Transcription;
RNA;
RNA Replicase;
Sensitivity and Specificity
- From:Annals of Laboratory Medicine
2017;37(6):494-498
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: Molecular detection of Middle East respiratory syndrome coronavirus (MERS-CoV) using real-time reverse transcription (rRT)-PCR assays is the method of choice for diagnosis of MERS. We evaluated the performance of the PowerChek MERS (upE & ORF1a) real-time PCR Kit (PowerChek MERS assay; Kogene Biotech, Korea) a one-step rRT-PCR assay for the qualitative detection of MERS-CoV. METHODS: We evaluated PowerChek MERS assay performance in comparison with nested RT-PCR and sequencing of the RNA-dependent RNA polymerase (RdRp) and N genes. To evaluate diagnostic sensitivity and specificity, 100 clinical specimens (50 positive and 50 negative for MERS-CoV) were simultaneously tested by using the PowerChek MERS and sequencing assays. Assay performance, including limit of detection and precision, was evaluated in vitro by using MERS-CoV RNA transcripts. Analytical specificity was evaluated with a diverse collection of 16 respiratory virus–positive clinical specimens and 14 respiratory bacterial isolates. RESULTS: The 95% limits of detection of the PowerChek MERS assay for the upE and the open rading frame (ORF)1a were 16.2 copies/µL and 8.2 copies/µL, respectively. No cross-reactivity was observed. The diagnostic sensitivity and specificity of the PowerChek MERS assay were both 100% (95% confidence interval, 91.1–100%). CONCLUSIONS: The PowerChek MERS assay is a straightforward and accurate assay for detecting MERS-CoV RNA. The assay will be a useful tool for the rapid diagnosis of MERS and could prove especially important for MERS outbreak control.
