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Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA.
Ann Lab Med. 2017 Nov; 37(6):494-498.AL

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.

Authors+Show Affiliations

Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. changski@skku.edu.Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

28840986

Citation

Huh, Hee Jae, et al. "Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA." Annals of Laboratory Medicine, vol. 37, no. 6, 2017, pp. 494-498.
Huh HJ, Kim JY, Kwon HJ, et al. Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA. Ann Lab Med. 2017;37(6):494-498.
Huh, H. J., Kim, J. Y., Kwon, H. J., Yun, S. A., Lee, M. K., Ki, C. S., Lee, N. Y., & Kim, J. W. (2017). Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA. Annals of Laboratory Medicine, 37(6), 494-498. https://doi.org/10.3343/alm.2017.37.6.494
Huh HJ, et al. Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA. Ann Lab Med. 2017;37(6):494-498. PubMed PMID: 28840986.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA. AU - Huh,Hee Jae, AU - Kim,Ji Youn, AU - Kwon,Hyeon Jeong, AU - Yun,Sun Ae, AU - Lee,Myoung Keun, AU - Ki,Chang Seok, AU - Lee,Nam Yong, AU - Kim,Jong Won, PY - 2017/02/03/received PY - 2017/03/26/revised PY - 2017/07/12/accepted PY - 2017/8/26/entrez PY - 2017/8/26/pubmed PY - 2018/5/12/medline KW - MERS-CoV KW - Performance KW - Real-time reverse-transcription PCR SP - 494 EP - 498 JF - Annals of laboratory medicine JO - Ann Lab Med VL - 37 IS - 6 N2 - 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. SN - 2234-3814 UR - https://www.unboundmedicine.com/medline/citation/28840986/Performance_Evaluation_of_the_PowerChek_MERS__upE_&_ORF1a__Real_Time_PCR_Kit_for_the_Detection_of_Middle_East_Respiratory_Syndrome_Coronavirus_RNA_ DB - PRIME DP - Unbound Medicine ER -