tetano
Editor, Senior Moderator
Clin Chem. 2020 Apr 4. pii: hvaa091. doi: 10.1093/clinchem/hvaa091. [Epub ahead of print]
Potential false-negative nucleic acid testing results for Severe Acute Respiratory Syndrome Coronavirus 2 from thermal inactivation of samples with low viral loads.
Pan Y[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Long L[SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP], Zhang D[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Yan T[SUP]7[/SUP], Cui S[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Yang P[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Wang Q[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Ren S[SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP].
Author information
Abstract
BACKGROUND:
Corona Virus 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 work 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.
? 2020 American Association for Clinical Chemistry.
KEYWORDS:
COVID-19; False negative; Nucleic Acid Test; RT-PCR; SARS-CoV-2; Thermal Inactivation
PMID:32246822DOI:10.1093/clinchem/hvaa091
Potential false-negative nucleic acid testing results for Severe Acute Respiratory Syndrome Coronavirus 2 from thermal inactivation of samples with low viral loads.
Pan Y[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Long L[SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP], Zhang D[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Yan T[SUP]7[/SUP], Cui S[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Yang P[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Wang Q[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP], Ren S[SUP]4,[/SUP][SUP]5,[/SUP][SUP]6[/SUP].
Author information
Abstract
BACKGROUND:
Corona Virus 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 work 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.
? 2020 American Association for Clinical Chemistry.
KEYWORDS:
COVID-19; False negative; Nucleic Acid Test; RT-PCR; SARS-CoV-2; Thermal Inactivation
PMID:32246822DOI:10.1093/clinchem/hvaa091