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胍盐-硫氰酸基于核酸提取缓冲液在潜在传染性材料中用于灭活脊髓灰质炎病毒。

Use of guanidine thiocyanate-based nucleic acid extraction buffers to inactivate poliovirus in potentially infectious materials.

机构信息

IHRC, Inc., Contracting Agency to the Centers for Disease Control and Prevention, Atlanta, GA, 30346, USA.

Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, 30329, USA.

出版信息

J Virol Methods. 2021 Nov;297:114262. doi: 10.1016/j.jviromet.2021.114262. Epub 2021 Aug 9.

Abstract

The efforts of the Global Poliovirus Eradication Initiative (GPEI) have brought about the near elimination of poliovirus worldwide. The World Health Organization has issued guidelines for the safe handling and containment of infectious materials (IM) and potentially infectious materials (PIM) following poliovirus eradication. Inactivation of poliovirus in IM and PIM is needed to prevent inadvertent re-introduction of polioviruses post-eradication. In this study, we investigated the use of guanidine thiocyanate-based nucleic acid extraction buffers from commercially available nucleic acid extraction kits to inactivate poliovirus in cell culture isolates and stool suspensions, two common types of poliovirus IM and PIM, respectively. Incubation with selected nucleic acid extraction buffers or extraction buffers supplemented with ethanol reduced the infectivity of high-titer wild poliovirus type 1 (WPV1), wild poliovirus type 3 (WPV3), Sabin 1 (SL1), and Sabin 3 (SL3) cell culture isolates below the limit of detection in CCID assays. Stool suspensions containing WPV1, WPV3, SL1, SL2, or SL3 were also inactivated by the extraction buffers tested. Blind passage of WPV1-spiked stool suspensions confirmed complete inactivation of WPV1 after incubation with extraction buffers. Moreover, treatment with a buffer consisting of 4 M guanidine thiocyanate with 30 % ethanol inactivated a high-titer WPV1 culture isolate and a WPV1-spiked stool suspension. Taken together, these results show that guanidine thiocyanate-based nucleic acid extraction buffers are an effective means of inactivating poliovirus IM and PIM, and thus will be instrumental in ensuring containment compliance and preventing potential re-emergence of contained polioviruses.

摘要

全球消灭脊灰病毒倡议(GPEI)的努力使脊灰病毒在全球几乎被消灭。世界卫生组织发布了关于安全处理和控制感染性材料(IM)和潜在感染性材料(PIM)的指南,以应对脊灰病毒的消除。需要对 IM 和 PIM 中的脊灰病毒进行灭活,以防止在消灭后无意中重新引入脊灰病毒。在这项研究中,我们研究了从市售核酸提取试剂盒中使用胍基硫氰酸盐基核酸提取缓冲液来灭活细胞培养物分离株和粪便悬浮液中的脊灰病毒,这两种分别是常见的脊灰病毒 IM 和 PIM 类型。用选定的核酸提取缓冲液或添加乙醇的提取缓冲液孵育可使高滴度野生脊灰病毒 1 型(WPV1)、野生脊灰病毒 3 型(WPV3)、Sabin1(SL1)和 Sabin3(SL3)细胞培养分离株的感染性在 CCID 测定中低于检测限。含 WPV1、WPV3、SL1、SL2 或 SL3 的粪便悬浮液也被测试的提取缓冲液灭活。WPV1 污染粪便悬浮液的盲传证实,用提取缓冲液孵育后 WPV1 完全失活。此外,用 4 M 胍基硫氰酸盐和 30%乙醇组成的缓冲液处理可灭活高滴度 WPV1 培养分离株和 WPV1 污染的粪便悬浮液。综上所述,这些结果表明胍基硫氰酸盐基核酸提取缓冲液是一种有效灭活脊灰病毒 IM 和 PIM 的方法,因此对于确保包容合规性和防止包含的脊灰病毒再次出现将是至关重要的。

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