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End-to-End Protocol for the Detection of SARS-CoV-2 from Built Environments

机译:从内置环境检测SARS-COV-2的端到端协议

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes coronavirus disease 2019, is a respiratory virus primarily transmitted person to person through inhalation of droplets or aerosols, laden with viral particles. However, as recent studies have shown, virions can remain infectious for up to 72?h on surfaces, which can lead to transmission through contact. Thus, a comprehensive study was conducted to determine the efficiency of protocols to recover SARS-CoV-2 from surfaces in built environments. This end-to-end (E2E) study showed that the effective combination for monitoring SARS-CoV-2 on surfaces includes using an Isohelix swab collection tool, DNA/RNA Shield as a preservative, an automated system for RNA extraction, and reverse transcriptase quantitative PCR (RT-qPCR) as the detection assay. Using this E2E approach, this study showed that, in some cases, noninfectious viral fragments of SARS-CoV-2 persisted on surfaces for as long as 8 days even after bleach treatment. Additionally, debris associated with specific built environment surfaces appeared to inhibit and negatively impact the recovery of RNA; Amerstat demonstrated the highest inhibition (90%) when challenged with an inactivated viral control. Overall, it was determined that this E2E protocol required a minimum of 1,000 viral particles per 25 cm 2 to successfully detect virus from test surfaces. Despite our findings of viral fragment longevity on surfaces, when this method was employed to evaluate 368 samples collected from various built environmental surfaces, all samples tested negative, indicating that the surfaces were either void of virus or below the detection limit of the assay. IMPORTANCE The ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (the virus responsible for coronavirus disease 2019 [COVID-19]) pandemic has led to a global slowdown with far-reaching financial and social impacts. The SARS-CoV-2 respiratory virus is primarily transmitted from person to person through inhalation of infected droplets or aerosols. However, some studies have shown that virions can remain infectious on surfaces for days and can lead to human infection from contact with infected surfaces. Thus, a comprehensive study was conducted to determine the efficiency of protocols to recover SARS-CoV-2 from surfaces in built environments. This end-to-end study showed that the effective combination for monitoring SARS-CoV-2 on surfaces required a minimum of 1,000 viral particles per 25 cm 2 to successfully detect virus from surfaces. This comprehensive study can provide valuable information regarding surface monitoring of various materials as well as the capacity to retain viral RNA and allow for effective disinfection.
机译:严重急性呼吸综合征冠状病毒2(SARS-COV-2),导致冠状病毒疾病2019年的病毒是一种呼吸道病毒,主要通过吸入液滴或气溶胶,载有病毒颗粒的液滴或气溶胶来传播人。然而,随着最近的研究表明,在表面上可以保持最高72Ω的病毒粒子,这可能导致通过接触传输。因此,进行了综合研究以确定从建造环境中的表面恢复SARS-COV-2的方案效率。这种端到端(E2E)的研究表明,在表面上监测SARS-COV-2的有效组合包括使用ISOHelix拭子收集工具,DNA / RNA屏蔽作为防腐剂,用于RNA提取的自动化系统和逆转录酶定量PCR(RT-QPCR)作为检测测定。使用这种E2E方法,本研究表明,在某些情况下,SARS-COV-2的非缺陷病毒片段甚至在漂白剂处理后甚至在8天内持续存在。另外,与特定构建环境表面相关的碎片似乎抑制和负面影响RNA的恢复;当用灭活病毒对照攻击时,Amstat展示了最高的抑制(> 90%)。总体而言,确定该E2E方案每25cm 2至少需要1,000个病毒颗粒,以从测试表面成功检测病毒。尽管研究了表面上的病毒片段寿命寿命寿命,但采用这种方法评估从各种建筑环境表面收集的368个样品,所有样品都测试为阴性,表明表面是病毒的空隙或低于测定的检测限。重要性持续的严重急性呼吸综合征Coronavirus 2(SARS-COV-2)(负责冠状病毒疾病的病毒2019年[Covid-19])大流行导致全球放缓,具有深远的财务和社会影响。 SARS-COV-2呼吸道病毒主要通过吸入感染的液滴或气溶胶来从人传播给人。然而,有些研究表明,病毒群体可以在几天内保持感染,并且可以导致人类​​感染与受感染的表面接触。因此,进行了综合研究以确定从建造环境中的表面恢复SARS-COV-2的方案效率。该端到端的研究表明,在表面上监测SARS-COV-2的有效组合需要每25cm 2最少的1,000个病毒颗粒以成功检测来自表面的病毒。该综合研究可以提供有关各种材料的表面监测的有价值的信息,以及保留病毒RNA的能力并允许有效消毒。
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