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Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus.
J Mol Diagn. 2017 11; 19(6):817-827.JM

Abstract

Middle East respiratory syndrome (MERS) is an emerging zoonotic viral respiratory disease that was first identified in Saudi Arabia in 2012. In 2015, the largest MERS outbreak outside of the Middle East region occurred in the Republic of Korea. The rapid nosocomial transmission of MERS-coronavirus (MERS-CoV) in Korean health care settings highlighted the importance and urgent need for a rapid and reliable on-site diagnostic assay to implement effective control and preventive measures. Here, the evaluation and validation of two newly developed reverse transcription-insulated isothermal PCR (RT-iiPCR) methods targeting the ORF1a and upE genes of MERS-CoV are described. Compared with World Health Organization-recommended singleplex real-time quantitative RT-PCR (RT-qPCR) assays, both RT-iiPCR assays had comparable analytical sensitivity for the detection of MERS-CoV RNA in tissue culture fluid and in sputum samples spiked with infectious virus. Furthermore, clinical evaluation was performed with sputum samples collected from subjects with acute and chronic respiratory illnesses, including patients infected with MERS-CoV. The overall agreement values between the two RT-iiPCR assays and the reference RT-qPCR assays were 98.06% (95% CI, 94.43%-100%; κ = 0.96) and 99.03% (95% CI, 95.88%-100%; κ = 0.99) for ORF1a and upE assays, respectively. The ORF1a and upE MERS-CoV RT-iiPCR assays coupled with a field-deployable system provide a platform for a highly sensitive and specific on-site tool for diagnosis of MERS-CoV infections.

Authors+Show Affiliations

Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon, Republic of Korea. Electronic address: yygo@krict.re.kr.Department of Medicine, Chungnam National University School of Medicine, Daejeon, Republic of Korea.Department of Medicine, Chungnam National University School of Medicine, Daejeon, Republic of Korea.Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon, Republic of Korea.Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea.Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon, Republic of Korea.Department of Microbiology and Immunology, and Department of Biomedical Science, Seoul National University College of Medicine and Bundang Hospital, Seoul, Republic of Korea.Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Republic of Korea.GeneReach USA, Lexington, Massachusetts.GeneReach USA, Lexington, Massachusetts.GeneReach USA, Lexington, Massachusetts.GeneReach USA, Lexington, Massachusetts.Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, College of Agriculture, Food and Environment and Department of Microbiology, Immunology and Molecular Genetics, College of Medicine, University of Kentucky, Lexington, Kentucky. Electronic address: ubalasuriya@uky.edu.

Pub Type(s)

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

Language

eng

PubMed ID

28807812

Citation

Go, Yun Young, et al. "Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus." The Journal of Molecular Diagnostics : JMD, vol. 19, no. 6, 2017, pp. 817-827.
Go YY, Kim YS, Cheon S, et al. Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus. J Mol Diagn. 2017;19(6):817-827.
Go, Y. Y., Kim, Y. S., Cheon, S., Nam, S., Ku, K. B., Kim, M., Cho, N. H., Park, H., Alison Lee, P. Y., Lin, Y. C., Tsai, Y. L., Thomas Wang, H. T., & Balasuriya, U. B. R. (2017). Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus. The Journal of Molecular Diagnostics : JMD, 19(6), 817-827. https://doi.org/10.1016/j.jmoldx.2017.06.007
Go YY, et al. Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus. J Mol Diagn. 2017;19(6):817-827. PubMed PMID: 28807812.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus. AU - Go,Yun Young, AU - Kim,Yeon-Sook, AU - Cheon,Shinhye, AU - Nam,Sangwoo, AU - Ku,Keun Bon, AU - Kim,Meehyein, AU - Cho,Nam Hyuk, AU - Park,Hyun, AU - Alison Lee,Pei-Yu, AU - Lin,Yu-Chun, AU - Tsai,Yun-Long, AU - Thomas Wang,Hwa-Tang, AU - Balasuriya,Udeni B R, Y1 - 2017/08/12/ PY - 2017/03/25/received PY - 2017/05/19/revised PY - 2017/06/21/accepted PY - 2017/8/16/pubmed PY - 2018/6/7/medline PY - 2017/8/16/entrez SP - 817 EP - 827 JF - The Journal of molecular diagnostics : JMD JO - J Mol Diagn VL - 19 IS - 6 N2 - Middle East respiratory syndrome (MERS) is an emerging zoonotic viral respiratory disease that was first identified in Saudi Arabia in 2012. In 2015, the largest MERS outbreak outside of the Middle East region occurred in the Republic of Korea. The rapid nosocomial transmission of MERS-coronavirus (MERS-CoV) in Korean health care settings highlighted the importance and urgent need for a rapid and reliable on-site diagnostic assay to implement effective control and preventive measures. Here, the evaluation and validation of two newly developed reverse transcription-insulated isothermal PCR (RT-iiPCR) methods targeting the ORF1a and upE genes of MERS-CoV are described. Compared with World Health Organization-recommended singleplex real-time quantitative RT-PCR (RT-qPCR) assays, both RT-iiPCR assays had comparable analytical sensitivity for the detection of MERS-CoV RNA in tissue culture fluid and in sputum samples spiked with infectious virus. Furthermore, clinical evaluation was performed with sputum samples collected from subjects with acute and chronic respiratory illnesses, including patients infected with MERS-CoV. The overall agreement values between the two RT-iiPCR assays and the reference RT-qPCR assays were 98.06% (95% CI, 94.43%-100%; κ = 0.96) and 99.03% (95% CI, 95.88%-100%; κ = 0.99) for ORF1a and upE assays, respectively. The ORF1a and upE MERS-CoV RT-iiPCR assays coupled with a field-deployable system provide a platform for a highly sensitive and specific on-site tool for diagnosis of MERS-CoV infections. SN - 1943-7811 UR - https://www.unboundmedicine.com/medline/citation/28807812/Evaluation_and_Clinical_Validation_of_Two_Field_Deployable_Reverse_Transcription_Insulated_Isothermal_PCR_Assays_for_the_Detection_of_the_Middle_East_Respiratory_Syndrome_Coronavirus_ DB - PRIME DP - Unbound Medicine ER -