An evaluation of a new rapid qPCR test for the detection of 2019-novel coronavirus nucleocapsid (N1) gene in wastewater in Roanoke and Salem VA sewersheds.

IF 2.5 4区 环境科学与生态学 Q3 ENVIRONMENTAL SCIENCES Journal of water and health Pub Date : 2024-08-01 DOI:10.2166/wh.2024.085
Lia Willow Lehrer, Anna Marie Lewis, Susan Tolliver, Marcia Degen, Rekha Singh, Sara Houser, Jayasimha Rao
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Abstract

The COVID-19 pandemic initiated public interest in wastewater-based epidemiology (WBE). Public and private entities responded to the need to produce timely and accurate data. LuminUltra and Hach partnered to provide a rapid, field-based quantitative polymerase chain reaction (qPCR) test for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wastewater. This study evaluates the Hach GeneCount SARS-CoV-2 Wastewater RT-qPCR Assay Kit and LuminUltra GeneCount® Q-16 RT-PCR instrument. The Hach LuminUltra methods were compared to the Promega Wizard® Enviro Total Nucleic Acid kit and Bio-Rad CFX Opus 96 Real-time PCR Detection System. Over a 12-week period, wastewater samples were collected weekly from seven locations in the Roanoke/Salem, VA sewersheds. Concentration and extraction of the viral RNA were followed by qPCR analysis. The target gene for detection was the nucleocapsid gene (N1) of the SARS-CoV-2 virus. Costs, ease of use, time to produce results, sample preparation, and data comparisons were considered. The comparison determined that the Hach LuminUltra method and instrument were more affordable, consumed less time, and required less technical expertise. While the new method was specific, it had low sensitivity. This evaluation suggests the Hach LuminUltra method should be reserved for limited situations requiring onsite field analysis where data accuracy is not essential.

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评估用于检测弗吉尼亚州罗诺克和塞勒姆下水道废水中 2019 年新型冠状病毒核头壳 (N1) 基因的新型快速 qPCR 检测方法。
COVID-19 大流行引发了公众对基于废水的流行病学 (WBE) 的兴趣。公共和私营机构纷纷响应及时提供准确数据的需求。LuminUltra 和 Hach 合作提供了一种基于现场的快速定量聚合酶链反应 (qPCR) 测试,用于检测废水中的严重急性呼吸系统综合征冠状病毒 2 (SARS-CoV-2)。这项研究评估了 Hach GeneCount SARS-CoV-2 废水 RT-qPCR 检测试剂盒和 LuminUltra GeneCount® Q-16 RT-PCR 仪器。Hach LuminUltra 方法与 Promega Wizard® Enviro 总核酸试剂盒和 Bio-Rad CFX Opus 96 实时 PCR 检测系统进行了比较。在为期 12 周的时间里,每周从弗吉尼亚州罗诺克/塞勒姆下水道的七个地点收集废水样本。浓缩和提取病毒 RNA 后进行 qPCR 分析。检测的目标基因是 SARS-CoV-2 病毒的核壳基因 (N1)。对成本、易用性、产生结果的时间、样品制备和数据比较进行了考虑。比较结果表明,Hach LuminUltra 方法和仪器更经济实惠,耗时更短,所需的专业技术知识也更少。虽然新方法具有特异性,但灵敏度较低。评估结果表明,Hach LuminUltra 方法只适用于需要现场实地分析的有限情况,数据准确性并不重要。
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来源期刊
Journal of water and health
Journal of water and health 环境科学-环境科学
CiteScore
3.60
自引率
8.70%
发文量
110
审稿时长
18-36 weeks
期刊介绍: Journal of Water and Health is a peer-reviewed journal devoted to the dissemination of information on the health implications and control of waterborne microorganisms and chemical substances in the broadest sense for developing and developed countries worldwide. This is to include microbial toxins, chemical quality and the aesthetic qualities of water.
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