Degradation of SARS-CoV-2 specific ribonucleic acid in samples for nucleic acid amplification detection

Katsuyuki Takeuchi,Yoritsugu Iida,Yukiko Kurosawa,Shigehiko Fujimaki,Hiroyuki Yanagisawa,Kosuke Kawai,Shuang-Yong Xu

doi:10.1371/journal.pone.0264541

Katsuyuki Takeuchi, Yoritsugu Iida + Show 5 more

Open Access

https://doi.org/10.1371/journal.pone.0264541

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Abstract

The degradation of SARS-CoV-2 specific ribonucleic acid (RNA) was investigated by a numerical modeling approach based on nucleic acid amplification test (NAAT) results utilizing the SmartAmp technique. The precision of the measurement was verified by the relative standard deviation (RSD) of repeated measurements at each calibration point. The precision and detection limits were found to be 6% RSD (seven repeated measurements) and 94 copies/tube, respectively, at the lowest calibration point. RNA degradation curves obtained from NAAT data on four different temperatures were in good agreement with the first-order reaction model. By referring to rate constants derived from the results, the Arrhenius model was applied to predict RNA degradation behavior. If the initial RNA concentration was high enough, such as in samples taken from infected bodies, the NAAT results were expected to be positive during testing. On the other hand, if initial RNA concentrations were relatively low, such as RNA in residual viruses on environmental surfaces, special attention should be paid to avoid false-negative results. The results obtained in this study provide a practical guide for RNA sample management in the NAAT of non-human samples.

Highlights

The viral infection coronavirus disease 2019 (COVID-19) appeared in late 2019 and by 2020 had been classified a pandemic
Arrhenius modeling indicates that higher temperature enhances the ribonucleic acid (RNA) degradation rate. Given such rapid RNA degradation under ambient temperature, our result provides a practical guide to RNA sample management in nucleic acid amplification test (NAAT) for residual viruses sampled from environmental surfaces under temperature-changing conditions
The RNA degradation curves obtained from the SmartAmp measurements on four different temperatures (4 ̊C, 25 ̊C, 37 ̊C, and 45 ̊C) were in good agreement with the first-order reaction model

Summary

Introduction

The viral infection coronavirus disease 2019 (COVID-19) appeared in late 2019 and by 2020 had been classified a pandemic. The COVID-19 pandemic claimed millions of lives and infected hundreds of millions of people worldwide [1]. Mass vaccination has mitigated COVID-19, yet SARS-CoV-2 variants have been discovered in many parts of the world [2–4]. The pandemic is still dominating the globe, posing a threat to the global economy by causing lockdowns in cities, which causes blockages in multilateral logistics [5]. No one knows when humanity will overcome COVID-19. The nucleic acid amplification test (NAAT) is an effective method for identifying COVID-infected individuals and enables positive patients to be transported to isolation or medical facilities for proper treatment [6, 7]

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