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Culture-Based Virus Isolation To Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19.
J Clin Microbiol. 2020 07 23; 58(8)JC

Abstract

Real-time reverse transcription-PCR (RT-PCR) is currently the most sensitive method to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes coronavirus disease 2019 (COVID-19). However, the correlation between detectable viral RNA and culturable virus in clinical specimens remains unclear. Here, we performed virus culture for 60 specimens that were confirmed to be positive for SARS-CoV-2 RNA by real-time RT-PCR. The virus could be successfully isolated from 12 throat and nine nasopharyngeal swabs and two sputum specimens. The lowest copy number required for virus isolation was determined to be 5.4, 6.0, and 5.7 log10 genome copies/ml sample for detecting the nsp12, E, and N genes, respectively. We further examined the correlation of genome copy number and virus isolation in different regions of the viral genome, demonstrating that culturable specimens are characterized by high copy numbers with a linear correlation observed between copy numbers of amplicons targeting structural and nonstructural regions. Overall, these results indicate that in addition to the copy number, the integrity of the viral genome should be considered when evaluating the infectivity of clinical SARS-CoV-2 specimens.

Authors+Show Affiliations

Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan. Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan. Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan.Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan. Division of Infectious Diseases, Department of Pediatrics, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan. Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan kctsao@cgmh.org.tw srshih@mail.cgu.edu.tw. Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan. Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan.Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan kctsao@cgmh.org.tw srshih@mail.cgu.edu.tw. Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, Taiwan. Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taoyuan, Taiwan. Research Center for Chinese Herbal Medicine, Research Center for Food and Cosmetic Safety, and Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, Taiwan.

Pub Type(s)

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

Language

eng

PubMed ID

32518072

Citation

Huang, Chung-Guei, et al. "Culture-Based Virus Isolation to Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19." Journal of Clinical Microbiology, vol. 58, no. 8, 2020.
Huang CG, Lee KM, Hsiao MJ, et al. Culture-Based Virus Isolation To Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19. J Clin Microbiol. 2020;58(8).
Huang, C. G., Lee, K. M., Hsiao, M. J., Yang, S. L., Huang, P. N., Gong, Y. N., Hsieh, T. H., Huang, P. W., Lin, Y. J., Liu, Y. C., Tsao, K. C., & Shih, S. R. (2020). Culture-Based Virus Isolation To Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19. Journal of Clinical Microbiology, 58(8). https://doi.org/10.1128/JCM.01068-20
Huang CG, et al. Culture-Based Virus Isolation to Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19. J Clin Microbiol. 2020 07 23;58(8) PubMed PMID: 32518072.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Culture-Based Virus Isolation To Evaluate Potential Infectivity of Clinical Specimens Tested for COVID-19. AU - Huang,Chung-Guei, AU - Lee,Kuo-Ming, AU - Hsiao,Mei-Jen, AU - Yang,Shu-Li, AU - Huang,Peng-Nien, AU - Gong,Yu-Nong, AU - Hsieh,Tzu-Hsuan, AU - Huang,Po-Wei, AU - Lin,Ya-Jhu, AU - Liu,Yi-Chun, AU - Tsao,Kuo-Chien, AU - Shih,Shin-Ru, Y1 - 2020/07/23/ PY - 2020/05/10/received PY - 2020/06/05/accepted PY - 2020/6/11/pubmed PY - 2020/8/5/medline PY - 2020/6/11/entrez KW - RT-PCR KW - SARS-CoV-2 KW - culturability KW - genome copy KW - genome integrity JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 58 IS - 8 N2 - Real-time reverse transcription-PCR (RT-PCR) is currently the most sensitive method to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes coronavirus disease 2019 (COVID-19). However, the correlation between detectable viral RNA and culturable virus in clinical specimens remains unclear. Here, we performed virus culture for 60 specimens that were confirmed to be positive for SARS-CoV-2 RNA by real-time RT-PCR. The virus could be successfully isolated from 12 throat and nine nasopharyngeal swabs and two sputum specimens. The lowest copy number required for virus isolation was determined to be 5.4, 6.0, and 5.7 log10 genome copies/ml sample for detecting the nsp12, E, and N genes, respectively. We further examined the correlation of genome copy number and virus isolation in different regions of the viral genome, demonstrating that culturable specimens are characterized by high copy numbers with a linear correlation observed between copy numbers of amplicons targeting structural and nonstructural regions. Overall, these results indicate that in addition to the copy number, the integrity of the viral genome should be considered when evaluating the infectivity of clinical SARS-CoV-2 specimens. SN - 1098-660X UR - https://www.unboundmedicine.com/medline/citation/32518072/Culture_Based_Virus_Isolation_To_Evaluate_Potential_Infectivity_of_Clinical_Specimens_Tested_for_COVID_19_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=32518072 DB - PRIME DP - Unbound Medicine ER -