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病毒干扰作为使用 BGM 细胞系的细胞培养-PCR 综合检测感染性腺病毒时出现假阴性的一个因素。

Viral Interference as a Factor of False-Negative in the Infectious Adenovirus Detection Using Integrated Cell Culture-PCR with a BGM Cell Line.

机构信息

Department of Frontier Sciences for Advanced Environment, Graduate School of Environmental Studies, Tohoku University, Aoba 6-6-06, Aramaki, Aoba-ku, Sendai, Miyagi, 980-8579, Japan.

Department of Civil and Environmental Engineering, Graduate School of Engineering, Tohoku University, Aoba 6-6-06, Aramaki, Aoba-ku, Sendai, Miyagi, 980-8579, Japan.

出版信息

Food Environ Virol. 2021 Mar;13(1):84-92. doi: 10.1007/s12560-020-09453-x. Epub 2021 Jan 4.

Abstract

This study investigated the influence of viral interference on the detection of enteric viruses using the integrated cell culture (ICC)-PCR with a BGM cell line. It was possible to detect 10 plaque-forming units (PFU)/flask of enterovirus 71 (EV71) in spite of the presence of 10 PFU/flask of adenovirus 40 (AdV40). Meanwhile, 10 PFU/flask of AdV40 was not detected in the presence of 10 PFU/flask of EV71. This inhibition of AdV40 detection using ICC-PCR was attributable to the growth of EV71, because the addition of a growth inhibitor of EV71 (rupintrivir) neutralized the detection inhibition of AdV40. The growth inhibition of AdV40 under co-infection with EV71 is probably caused by the immune responses of EV71-infected cells. AdV is frequently used as a fecal contamination indicator of environmental water, but this study demonstrated that false-negative detection of infectious AdV using ICC-PCR could be caused by the co-existence of infectious EV in a water sample. The addition of rupintrivir could prevent false-negative detection of AdV using ICC-PCR. This study, therefore, emphasizes the importance of confirming the presence of multiple enteric viruses in a sample derived from environmental water prior to the application of ICC-PCR because the viral interference phenomenon may lead to the false-negative detection of target viruses.

摘要

本研究探讨了病毒干扰对使用整合细胞培养(ICC)-PCR 与 BGM 细胞系检测肠道病毒的影响。尽管存在 10 噬菌斑形成单位(PFU)/瓶的腺病毒 40(AdV40),仍有可能检测到 10 PFU/瓶的肠道病毒 71(EV71)。同时,在存在 10 PFU/瓶 EV71 的情况下,未检测到 10 PFU/瓶的 AdV40。这种 ICC-PCR 检测 AdV40 的抑制作用归因于 EV71 的生长,因为添加 EV71 的生长抑制剂(rupintrivir)中和了 AdV40 的检测抑制作用。在与 EV71 共同感染下,AdV40 的生长抑制可能是由 EV71 感染细胞的免疫反应引起的。AdV 常用于环境水样中粪便污染的指示物,但本研究表明,ICC-PCR 检测传染性 AdV 可能会因水样中存在传染性 EV 而导致假阴性。添加 rupintrivir 可以防止 ICC-PCR 检测 AdV 的假阴性。因此,本研究强调在应用 ICC-PCR 之前,确认来自环境水样的样本中存在多种肠道病毒的重要性,因为病毒干扰现象可能导致目标病毒的假阴性检测。

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