Application of Capsid Integrity (RT-)qPCR to Assessing Occurrence of Intact Viruses in Surface Water and Tap Water in Japan
Application of Capsid Integrity (RT-)qPCR to Assessing Occurrence of Intact Viruses in Surface Water and Tap Water in Japan
复制标题
应用衣壳完整性 (RT-)qPCR 评估日本地表水和自来水中完整病毒的出现情况
DOI：
10.1016/j.watres.2020.116674
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发表时间：
2021
期刊：
影响因子：
12.8
通讯作者：
Katayama Hiroyuki
中科院分区：
文献类型：
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作者：
Canh Vu Duc;Torii Shotaro;Furumai Hiroaki;Katayama Hiroyuki
衣壳完整性（RT-）qPCR最近已被开发用于区分病毒的完整形式和灭活形式，但其用于鉴定饮用水中病毒完整性的适用性仍然有限。在这项研究中，我们研究了应用衣壳完整性（RT-）qPCR使用顺式二氯二氨铂（CDDP）与脱氧胆酸钠（SD）预处理（SD-CDDP-（RT-）qPCR）检测地表水和自来水中的完整病毒。共抽取63个水样（地表水，n = 20;自来水，n = 43），并通过常规（RT）-qPCR和SD-CDDP-（RT-）qPCR对辣椒温和斑驳病毒（PMMoV）和其他七种对人类致病的病毒进行定量（艾奇病毒（AiV）、基因型I和II的诺如病毒、肠病毒、腺病毒40和41型以及JC和BK多瘤病毒）。在地表水中，PMMoV（100%）比其他人类致病性病毒（30%-60%）更频繁地检测到，如通过常规（RT-）qPCR所确定的。SD-CDDP-（RT-）qPCR还显示，完整的PMMoV（95%）比完整的人类致病病毒（20%-45%）更常见。在自来水样品中，除了PMMoV（9%）和AiV（5%）外，大多数目标病毒均未被常规（RT-）qPCR检测到。PMMoV保持阳性（5%），而当通过SD-CDDP-（RT-）qPCR测试时未检测到AiV，表明一些PMMoV具有完整的衣壳，而AiV具有受损的衣壳。从地下水中产生的自来水中存在AiV而不存在PMMoV，这可能表明PMMoV作为地下水中病毒指示剂的局限性。除了在地表水中大量存在外，还在自来水中检测到PMMoV，包括具有完整衣壳的PMMoV。因此，不存在完整的PMMoV可以用来保证从地表水生产的自来水的病毒安全性。
Capsid integrity (RT-)qPCR has recently been developed to discriminate between intact forms from inactivated forms of viruses, but its applicability to identifying integrity of viruses in drinking water has remained limited. In this study, we investigated the application of capsid integrity (RT-)qPCR using cis-dichlorodiammineplatinum (CDDP) with sodium deoxycholate (SD) pretreatment (SD-CDDP-(RT-)qPCR) to detect intact viruses in surface water and tap water. A total of 63 water samples (surface water, n = 20; tap water, n = 43) were collected in the Kanto region in Japan and quantified by conventional (RT)-qPCR and SD-CDDP-(RT-)qPCR for pepper mild mottle virus (PMMoV) and seven other viruses pathogenic to humans (Aichivirus (AiV), noroviruses of genotypes I and II, enterovirus, adenovirus type 40 and 41, and JC and BK polyomaviruses). In surface water, PMMoV (100%) was more frequently detected than other human pathogenic viruses (30%–60%), as determined by conventional (RT-)qPCR. SD-CDDP-(RT-)qPCR also revealed that intact PMMoV (95%) was more common than intact human pathogenic viruses (20%–45%). In the tap water samples, most of the target viruses were not detected by conventional (RT-)qPCR, except for PMMoV (9%) and AiV (5%). PMMoV remained positive (5%), whereas no AiV was detected when tested by SD-CDDP-(RT-)qPCR, indicating that some PMMoV had an intact capsid, whereas AiV had damaged capsids. The presence of AiV in the absence of PMMoV in tap water produced from groundwater may demonstrate the limitation of PMMoV as a viral indicator in groundwater. In addition to being abundant in surface water, PMMoV was detected in tap water, including PMMoV with intact capsids. Thus, the absence of intact PMMoV may be used to guarantee the viral safety of tap water produced from surface water.