The effect of heat-treatment on SARS-CoV-2 viability and detection.
The effect of heat-treatment on SARS-CoV-2 viability and detection.
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DOI：
10.1016/j.jviromet.2021.114087
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发表时间：
2021-04
影响因子：
3.1
通讯作者：
Roberts ADG
中科院分区：
文献类型：
--
作者：
Burton J;Love H;Richards K;Burton C;Summers S;Pitman J;Easterbrook L;Davies K;Spencer P;Killip M;Cane P;Bruce C;Roberts ADG
为在生物安全等级2（BSL 2）下使用SARS-CoV-2临床样本开发安全的诊断方案需要了解热处理对SARS-CoV-2活性和下游RT-PCR灵敏度的影响。在这项研究中，将SARS-CoV-2/England/2/2020加热至56 °C和60 °C持续15、30和60分钟可使病毒滴度降低2.1至4.9 log 10 pfu/mL（通过空斑试验测定）。未发生完全灭活，重复样品之间存在显著变异性。在80 °C下热处理15或30 min后，通过空斑试验检测到活病毒，但60或90 min未检测到。在80 °C下热处理60 min后，仅通过更敏感的病毒培养物检测到感染性病毒。加热至80 °C 90 min或95 °C 1或5 min后未检测到活病毒。加热至56 °C和60 °C不会损害RT-PCR灵敏度。然而，在将病毒加热至80 °C 30 min或更长时间，或95 °C 1或5 min后，RT-PCR灵敏度降低（Ct值增加≥3）。总之，我们发现热灭活的效果随温度和持续时间的不同而变化很大。因此，热灭活及其下游效应的局部验证对于分子检测至关重要。
The development of safe diagnostic protocols for working with SARS-CoV-2 clinical samples at Biosafety Level 2 (BSL2) requires understanding of the effect of heat-treatment on SARS-CoV-2 viability and downstream RT-PCR sensitivity. In this study heating SARS-CoV-2/England/2/2020 to 56 °C and 60 °C for 15, 30 and 60 min reduced the virus titre by between 2.1 and 4.9 log10 pfu/mL (as determined by plaque assay). Complete inactivation did not occur and there was significant variability between replicates. Viable virus was detected by plaque assay after heat-treatment at 80 °C for 15 or 30 min but not 60 or 90 min. After heat-treatment at 80 °C for 60 min infectious virus was only detected by more sensitive virus culture. No viable virus was detected after heating to 80 °C for 90 min or 95 °C for 1 or 5 min. RT-PCR sensitivity was not compromised by heating to 56 °C and 60 °C. However, RT-PCR sensitivity was reduced (≥3 Ct value increase) after heating the virus to 80 °C for 30 min or longer, or 95 °C for 1 or 5 min. In summary we found that the efficacy of heat-inactivation varies greatly depending on temperature and duration. Local validation of heat-inactivation and its effects downstream is therefore essential for molecular testing.
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影响因子：
16.6
作者：
Smyrlaki I;Ekman M;Lentini A;Rufino de Sousa N;Papanicolaou N;Vondracek M;Aarum J;Safari H;Muradrasoli S;Rothfuchs AG;Albert J;Högberg B;Reinius B
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影响因子：
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影响因子：
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影响因子：
5.4
作者：
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通讯作者：
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