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Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR.
Ann Lab Med. 2016 Sep; 36(5):450-6.AL

Abstract

BACKGROUND

During the 2015 outbreak of Middle East Respiratory Syndrome coronavirus (MERS-CoV), six different commercial MERS-CoV RNA detection kits based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) were available in Korea. We performed analytical and clinical validations of these kits.

METHODS

PowerChek (Kogene Biotech, Korea), DiaPlexQ (SolGent, Korea), Anyplex (Seegene, Korea), AccuPower (Bioneer, Korea), LightMix (Roche Molecular Diagnostics, Switzerland), and UltraFast kits (Nanobiosys, Korea) were evaluated. Limits of detection (LOD) with 95% probability values were estimated by testing 16 replicates of upstream of the envelope gene (upE) and open reading frame 1a (ORF1a) RNA transcripts. Specificity was estimated by using 28 nasopharyngeal swabs that were positive for other respiratory viruses. Clinical sensitivity was evaluated by using 18 lower respiratory specimens. The sensitivity test panel and the high inhibition panel were composed of nine specimens each, including eight and six specimens that were positive for MERS-CoV, respectively.

RESULTS

The LODs for upE ranged from 21.88 to 263.03 copies/reaction, and those for ORF1a ranged from 6.92 to 128.82 copies/reaction. No cross-reactivity with other respiratory viruses was found. All six kits correctly identified 8 of 8 (100%) positive clinical specimens. Based on results from the high inhibition panel, PowerChek and AccuPower were the least sensitive to the presence of PCR inhibition.

CONCLUSIONS

The overall sensitivity and specificity of all six assay systems were sufficient for diagnosing MERS-CoV infection. However, the analytical sensitivity and detection ability in specimens with PCR inhibition could be improved with the use of appropriate internal controls.

Authors+Show Affiliations

Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea. mnkim@amc.seoul.kr.Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.Department of Laboratory Medicine, Hallym University College of Medicine, Hallym University Kangdong Sacred Heart Hospital, Seoul, Korea.Department of Laboratory Medicine, Inje University, Sanggye Paik Hospital, Seoul, Korea.Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

27374710

Citation

Kim, Mi Na, et al. "Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based On Real-Time Reverse-Transcription PCR." Annals of Laboratory Medicine, vol. 36, no. 5, 2016, pp. 450-6.
Kim MN, Ko YJ, Seong MW, et al. Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR. Ann Lab Med. 2016;36(5):450-6.
Kim, M. N., Ko, Y. J., Seong, M. W., Kim, J. S., Shin, B. M., & Sung, H. (2016). Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR. Annals of Laboratory Medicine, 36(5), 450-6. https://doi.org/10.3343/alm.2016.36.5.450
Kim MN, et al. Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based On Real-Time Reverse-Transcription PCR. Ann Lab Med. 2016;36(5):450-6. PubMed PMID: 27374710.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR. AU - Kim,Mi Na, AU - Ko,Young Jin, AU - Seong,Moon Woo, AU - Kim,Jae Seok, AU - Shin,Bo Moon, AU - Sung,Heungsup, PY - 2015/09/22/received PY - 2016/02/27/revised PY - 2016/05/16/accepted PY - 2016/7/5/entrez PY - 2016/7/5/pubmed PY - 2017/2/9/medline KW - Commercial kits KW - Lower respiratory specimen KW - MERS coronavirus KW - Real-time reverse transcription polymerase chain reaction KW - Validation SP - 450 EP - 6 JF - Annals of laboratory medicine JO - Ann Lab Med VL - 36 IS - 5 N2 - BACKGROUND: During the 2015 outbreak of Middle East Respiratory Syndrome coronavirus (MERS-CoV), six different commercial MERS-CoV RNA detection kits based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) were available in Korea. We performed analytical and clinical validations of these kits. METHODS: PowerChek (Kogene Biotech, Korea), DiaPlexQ (SolGent, Korea), Anyplex (Seegene, Korea), AccuPower (Bioneer, Korea), LightMix (Roche Molecular Diagnostics, Switzerland), and UltraFast kits (Nanobiosys, Korea) were evaluated. Limits of detection (LOD) with 95% probability values were estimated by testing 16 replicates of upstream of the envelope gene (upE) and open reading frame 1a (ORF1a) RNA transcripts. Specificity was estimated by using 28 nasopharyngeal swabs that were positive for other respiratory viruses. Clinical sensitivity was evaluated by using 18 lower respiratory specimens. The sensitivity test panel and the high inhibition panel were composed of nine specimens each, including eight and six specimens that were positive for MERS-CoV, respectively. RESULTS: The LODs for upE ranged from 21.88 to 263.03 copies/reaction, and those for ORF1a ranged from 6.92 to 128.82 copies/reaction. No cross-reactivity with other respiratory viruses was found. All six kits correctly identified 8 of 8 (100%) positive clinical specimens. Based on results from the high inhibition panel, PowerChek and AccuPower were the least sensitive to the presence of PCR inhibition. CONCLUSIONS: The overall sensitivity and specificity of all six assay systems were sufficient for diagnosing MERS-CoV infection. However, the analytical sensitivity and detection ability in specimens with PCR inhibition could be improved with the use of appropriate internal controls. SN - 2234-3814 UR - https://www.unboundmedicine.com/medline/citation/27374710/Analytical_and_Clinical_Validation_of_Six_Commercial_Middle_East_Respiratory_Syndrome_Coronavirus_RNA_Detection_Kits_Based_on_Real_Time_Reverse_Transcription_PCR_ DB - PRIME DP - Unbound Medicine ER -