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External Quality Assessment of MERS-CoV Molecular Diagnostics During the 2015 Korean Outbreak.
Ann Lab Med. 2016 May; 36(3):230-4.AL

Abstract

BACKGROUND

The largest outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) infection outside Middle East Asia in 2015 has necessitated the rapid expansion of laboratories that conduct MERS-CoV molecular testing in Korea, together with external quality assessment (EQA) to evaluate the assays used.

METHODS

The EQA program consisted of two phases; self-validation and blind assessment. For the first EQA phase, in vitro transcribed upstream region of the envelope gene (upE) and the open reading frame (ORF)1a RNAs were used at a concentration of 1,000 copies/μL. The test panel for the second EQA phase consisted of RNA extracts from three samples, which were obtained from two MERS-CoV positive patients and one MERS-CoV negative patient.

RESULTS

The first EQA phase results for 46 participants showed a linear relationship between the threshold cycle (C(T)) values of RNA materials and the logarithmic concentrations for both upE and ORF1a gene targets (R²=0.73 and 0.75, respectively). The mean C(T) value for each concentration was different depending on which commercial kit was used for the assay. Among the three commonly used kits, PowerChek MERS Real-Time PCR kit (KogeneBiotech, Korea) showed the lowest C(T) values at all concentrations of upE and most concentrations of ORF1a. The second EQA phase results for 47 participants were 100% correct for all tested samples.

CONCLUSIONS

This EQA survey demonstrates that the MERS-CoV molecular testing performed in Korea during the 2015 outbreak is of robust capability. However, careful establishment and validation of a cut-off value are recommended to ensure good analytical sensitivity.

Authors+Show Affiliations

Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.Department of Laboratory Medicine, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea.Department of Laboratory Medicine, Kangdong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea. sparkle@snu.ac.kr.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26915611

Citation

Seong, Moon Woo, et al. "External Quality Assessment of MERS-CoV Molecular Diagnostics During the 2015 Korean Outbreak." Annals of Laboratory Medicine, vol. 36, no. 3, 2016, pp. 230-4.
Seong MW, Lee SJ, Cho SI, et al. External Quality Assessment of MERS-CoV Molecular Diagnostics During the 2015 Korean Outbreak. Ann Lab Med. 2016;36(3):230-4.
Seong, M. W., Lee, S. J., Cho, S. I., Ko, K., Kim, M. N., Sung, H., Kim, J. S., Ahn, J. S., Yu, B. S., Kim, T. S., Kim, E. C., & Park, S. S. (2016). External Quality Assessment of MERS-CoV Molecular Diagnostics During the 2015 Korean Outbreak. Annals of Laboratory Medicine, 36(3), 230-4. https://doi.org/10.3343/alm.2016.36.3.230
Seong MW, et al. External Quality Assessment of MERS-CoV Molecular Diagnostics During the 2015 Korean Outbreak. Ann Lab Med. 2016;36(3):230-4. PubMed PMID: 26915611.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - External Quality Assessment of MERS-CoV Molecular Diagnostics During the 2015 Korean Outbreak. AU - Seong,Moon Woo, AU - Lee,Seung Jun, AU - Cho,Sung Im, AU - Ko,Kyungphil, AU - Kim,Mi Na, AU - Sung,Heungsub, AU - Kim,Jae Seok, AU - Ahn,Ji Soo, AU - Yu,Byung Su, AU - Kim,Taek Soo, AU - Kim,Eui Chong, AU - Park,Sung Sup, PY - 2015/09/04/received PY - 2015/11/11/revised PY - 2016/01/29/accepted PY - 2016/2/27/entrez PY - 2016/2/27/pubmed PY - 2017/1/10/medline KW - External quality assessment KW - MERS-CoV KW - Molecular diagnostics KW - ORF1a KW - Real-time PCR KW - upE SP - 230 EP - 4 JF - Annals of laboratory medicine JO - Ann Lab Med VL - 36 IS - 3 N2 - BACKGROUND: The largest outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) infection outside Middle East Asia in 2015 has necessitated the rapid expansion of laboratories that conduct MERS-CoV molecular testing in Korea, together with external quality assessment (EQA) to evaluate the assays used. METHODS: The EQA program consisted of two phases; self-validation and blind assessment. For the first EQA phase, in vitro transcribed upstream region of the envelope gene (upE) and the open reading frame (ORF)1a RNAs were used at a concentration of 1,000 copies/μL. The test panel for the second EQA phase consisted of RNA extracts from three samples, which were obtained from two MERS-CoV positive patients and one MERS-CoV negative patient. RESULTS: The first EQA phase results for 46 participants showed a linear relationship between the threshold cycle (C(T)) values of RNA materials and the logarithmic concentrations for both upE and ORF1a gene targets (R²=0.73 and 0.75, respectively). The mean C(T) value for each concentration was different depending on which commercial kit was used for the assay. Among the three commonly used kits, PowerChek MERS Real-Time PCR kit (KogeneBiotech, Korea) showed the lowest C(T) values at all concentrations of upE and most concentrations of ORF1a. The second EQA phase results for 47 participants were 100% correct for all tested samples. CONCLUSIONS: This EQA survey demonstrates that the MERS-CoV molecular testing performed in Korea during the 2015 outbreak is of robust capability. However, careful establishment and validation of a cut-off value are recommended to ensure good analytical sensitivity. SN - 2234-3814 UR - https://www.unboundmedicine.com/medline/citation/26915611/External_Quality_Assessment_of_MERS_CoV_Molecular_Diagnostics_During_the_2015_Korean_Outbreak_ DB - PRIME DP - Unbound Medicine ER -