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Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 from Thermal Inactivation of Samples with Low Viral Loads.
Clin Chem. 2020 06 01; 66(6):794-801.CC

Abstract

BACKGROUND

Coronavirus disease-2019 (COVID-19) has spread widely throughout the world since the end of 2019. Nucleic acid testing (NAT) has played an important role in patient diagnosis and management of COVID-19. In some circumstances, thermal inactivation at 56°C has been recommended to inactivate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) before NAT. However, this procedure could theoretically disrupt nucleic acid integrity of this single-stranded RNA virus and cause false negatives in real-time polymerase chain reaction (RT-PCR) tests.

METHODS

We investigated whether thermal inactivation could affect the results of viral NAT. We examined the effects of thermal inactivation on the quantitative RT-PCR results of SARS-CoV-2, particularly with regard to the rates of false-negative results for specimens carrying low viral loads. We additionally investigated the effects of different specimen types, sample preservation times, and a chemical inactivation approach on NAT.

RESULTS

Our study showed increased Ct values in specimens from diagnosed COVID-19 patients in RT-PCR tests after thermal incubation. Moreover, about half of the weak-positive samples (7 of 15 samples, 46.7%) were RT-PCR negative after heat inactivation in at least one parallel testing. The use of guanidinium-based lysis for preservation of these specimens had a smaller impact on RT-PCR results with fewer false negatives (2 of 15 samples, 13.3%) and significantly less increase in Ct values than heat inactivation.

CONCLUSION

Thermal inactivation adversely affected the efficiency of RT-PCR for SARS-CoV-2 detection. Given the limited applicability associated with chemical inactivators, other approaches to ensure the overall protection of laboratory personnel need consideration.

Authors+Show Affiliations

Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Prevention and Control, Beijing, P. R. China. Research Centre for Preventive Medicine of Beijing, Beijing, China. School of Public Health, Capital Medical University, Beijing, China.National Center for Clinical Laboratories, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, China. Graduate School, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China. Beijing Engineering Research Center of Laboratory Medicine, Beijing Hospital, Beijing, China.Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Prevention and Control, Beijing, P. R. China. Research Centre for Preventive Medicine of Beijing, Beijing, China. School of Public Health, Capital Medical University, Beijing, China.Department of Clinical Laboratory, Beijing Hospital, National Center of Gerontology, Beijing, China.Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Prevention and Control, Beijing, P. R. China. Research Centre for Preventive Medicine of Beijing, Beijing, China. School of Public Health, Capital Medical University, Beijing, China.Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Prevention and Control, Beijing, P. R. China. Research Centre for Preventive Medicine of Beijing, Beijing, China. School of Public Health, Capital Medical University, Beijing, China.Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Prevention and Control, Beijing, P. R. China. Research Centre for Preventive Medicine of Beijing, Beijing, China. School of Public Health, Capital Medical University, Beijing, China.National Center for Clinical Laboratories, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, China. Graduate School, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China. Beijing Engineering Research Center of Laboratory Medicine, Beijing Hospital, Beijing, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32246822

Citation

Pan, Yang, et al. "Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 From Thermal Inactivation of Samples With Low Viral Loads." Clinical Chemistry, vol. 66, no. 6, 2020, pp. 794-801.
Pan Y, Long L, Zhang D, et al. Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 from Thermal Inactivation of Samples with Low Viral Loads. Clin Chem. 2020;66(6):794-801.
Pan, Y., Long, L., Zhang, D., Yuan, T., Cui, S., Yang, P., Wang, Q., & Ren, S. (2020). Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 from Thermal Inactivation of Samples with Low Viral Loads. Clinical Chemistry, 66(6), 794-801. https://doi.org/10.1093/clinchem/hvaa091
Pan Y, et al. Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 From Thermal Inactivation of Samples With Low Viral Loads. Clin Chem. 2020 06 1;66(6):794-801. PubMed PMID: 32246822.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Potential False-Negative Nucleic Acid Testing Results for Severe Acute Respiratory Syndrome Coronavirus 2 from Thermal Inactivation of Samples with Low Viral Loads. AU - Pan,Yang, AU - Long,Luyao, AU - Zhang,Daitao, AU - Yuan,Tingting, AU - Cui,Shujuan, AU - Yang,Peng, AU - Wang,Quanyi, AU - Ren,Simei, PY - 2020/03/06/received PY - 2020/03/23/accepted PY - 2020/4/5/pubmed PY - 2020/6/17/medline PY - 2020/4/5/entrez KW - COVID-19 KW - RT-PCR KW - SARS-CoV-2 KW - false negative KW - nucleic acid test KW - thermal inactivation SP - 794 EP - 801 JF - Clinical chemistry JO - Clin Chem VL - 66 IS - 6 N2 - BACKGROUND: Coronavirus disease-2019 (COVID-19) has spread widely throughout the world since the end of 2019. Nucleic acid testing (NAT) has played an important role in patient diagnosis and management of COVID-19. In some circumstances, thermal inactivation at 56°C has been recommended to inactivate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) before NAT. However, this procedure could theoretically disrupt nucleic acid integrity of this single-stranded RNA virus and cause false negatives in real-time polymerase chain reaction (RT-PCR) tests. METHODS: We investigated whether thermal inactivation could affect the results of viral NAT. We examined the effects of thermal inactivation on the quantitative RT-PCR results of SARS-CoV-2, particularly with regard to the rates of false-negative results for specimens carrying low viral loads. We additionally investigated the effects of different specimen types, sample preservation times, and a chemical inactivation approach on NAT. RESULTS: Our study showed increased Ct values in specimens from diagnosed COVID-19 patients in RT-PCR tests after thermal incubation. Moreover, about half of the weak-positive samples (7 of 15 samples, 46.7%) were RT-PCR negative after heat inactivation in at least one parallel testing. The use of guanidinium-based lysis for preservation of these specimens had a smaller impact on RT-PCR results with fewer false negatives (2 of 15 samples, 13.3%) and significantly less increase in Ct values than heat inactivation. CONCLUSION: Thermal inactivation adversely affected the efficiency of RT-PCR for SARS-CoV-2 detection. Given the limited applicability associated with chemical inactivators, other approaches to ensure the overall protection of laboratory personnel need consideration. SN - 1530-8561 UR - https://www.unboundmedicine.com/medline/citation/32246822/Potential_False_Negative_Nucleic_Acid_Testing_Results_for_Severe_Acute_Respiratory_Syndrome_Coronavirus_2_from_Thermal_Inactivation_of_Samples_with_Low_Viral_Loads_ L2 - https://academic.oup.com/clinchem/article-lookup/doi/10.1093/clinchem/hvaa091 DB - PRIME DP - Unbound Medicine ER -