Prolonged Unfrozen Storage and Repeated Freeze-Thawing of SARS-CoV-2 Patient Samples Have Minor Effects on SARS-CoV-2 Detectability by RT-PCR.
Prolonged Unfrozen Storage and Repeated Freeze-Thawing of SARS-CoV-2 Patient Samples Have Minor Effects on SARS-CoV-2 Detectability by RT-PCR.
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DOI：
10.1016/j.jmoldx.2021.03.003
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发表时间：
2021-06
期刊：
影响因子：
--
通讯作者：
Bosshard PP
中科院分区：
文献类型：
--
作者：
Dzung A;Cheng PF;Stoffel C;Tastanova A;Turko P;Levesque MP;Bosshard PP
将严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）患者样本从擦拭站可靠地运送到诊断设施对于获得准确结果至关重要。因此，在运输时间较长的情况下，建议冷却或冷冻样品。在这项研究中，SARS-CoV-2的RT-PCR检测SARS-CoV-2的检测能力进行了评估后，长期解冻储存或反复冻融的SARS-CoV-2样本。将储存在病毒转运培养基中的SARS-CoV-2阳性患者拭子暴露于不同温度（4°C、25°C和35°C）和重复冻融，以评估储存条件对RT-PCR检测的影响。在病毒转运培养基中储存21天后，即使样品储存在35°C下，RT-PCR仍然可以可靠地检测到SARS-CoV-2 RNA。在35°C下，每天观察到的循环阈值的最大变化为0.046（±0.019），并且每天每个冻融循环观察到的循环阈值的最大变化为0.197（±0.06）。与在4°C下储存相比，在25°C或35°C下储存或反复冻融后，病毒RNA水平变化很小但显著。本研究的结果表明，病毒RNA水平在较高温度和反复冻融下相对稳定。
Reliable transportation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) patient samples from a swabbing station to a diagnostics facility is essential for accurate results. Therefore, cooling or freezing the samples is recommended in case of longer transportation times. In this study, SARS-CoV-2 detectability by RT-PCR was assessed after prolonged unfrozen storage or repetitive freeze-thawing of SARS-CoV-2 samples. SARS-CoV-2–positive patient swabs stored in viral transport medium were exposed to different temperatures (4°C, 25°C, and 35°C) and to repetitive freeze-thawing, to assess the effect of storage conditions on RT-PCR detection. SARS-CoV-2 RNA was still reliably detected by RT-PCR after 21 days of storage in viral transport medium, even when the samples had been stored at 35°C. The maximum observed change in cycle threshold value per day was 0.046 (±0.019) at 35°C, and the maximum observed change in cycle threshold value per freeze-thaw cycle per day was 0.197 (±0.06). Compared with storage at 4°C, viral RNA levels deviated little but significantly when stored at 25°C or 35°C, or after repeated freeze-thawing. The results of this study indicate that viral RNA levels are relatively stable at higher temperatures and repetitive freeze-thawing.
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DOI：
10.1016/j.jmoldx.2021.04.009
发表时间：
2021-07
期刊：
The Journal of molecular diagnostics : JMD
影响因子：
--
作者：
Tastanova A;Stoffel CI;Dzung A;Cheng PF;Bellini E;Johansen P;Duda A;Nobbe S;Lienhard R;Bosshard PP;Levesque MP
通讯作者：
Levesque MP
影响因子：
7.3
作者：
Abdelrahman Z;Li M;Wang X
通讯作者：
Wang X
影响因子：
--
作者：
D'Arienzo, Marco;Coniglio, Angela
通讯作者：
Coniglio, Angela
影响因子：
9.4
作者：
Druce, Julian;Garcia, Katherine;Birch, Chris
通讯作者：
Birch, Chris
影响因子：
5.8
作者：
Bates, Douglas;Maechler, Martin;Walker, Steven C.
通讯作者：
Walker, Steven C.