Evaluation of different molecular systems for detection and quantification of SARS-CoV-2 RNA from wastewater samples

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2024-05-23 DOI:10.1016/j.jviromet.2024.114956

Fabio Morecchiato , Marco Coppi , Claudia Niccolai , Alberto Antonelli , Leandro Di Gloria , Piergiuseppe Calà , Fabrizio Mancuso , Matteo Ramazzotti , Tommaso Lotti , Claudio Lubello , Gian Maria Rossolini

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引用次数: 0

Abstract

Wastewater-based epidemiology has proved to be a suitable approach for tracking the spread of epidemic agents including SARS-CoV-2 RNA. Different protocols have been developed for quantitative detection of SARS-CoV-2 RNA from wastewater samples, but little is known on their performance. In this study we compared three protocols based on Reverse Transcription Real Time-PCR (RT-PCR) and one based on Droplet Digital PCR (ddPCR) for SARS-CoV-2 RNA detection from 35 wastewater samples. Overall, SARS-CoV-2 RNA was detected by at least one method in 85.7 % of samples, while 51.4 %, 22.8 % and 8.6 % resulted positive with two, three or all four methods, respectively. Protocols based on commercial RT-PCR assays and on Droplet Digital PCR showed an overall higher sensitivity vs. an in-house assay. The use of more than one system, targeting different genes, could be helpful to increase detection sensitivity.

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评估用于检测和定量废水样本中 SARS-CoV-2 RNA 的不同分子系统

事实证明，以废水为基础的流行病学是追踪包括 SARS-CoV-2 RNA 在内的流行病病原体传播的合适方法。目前已开发出不同的方案来定量检测废水样本中的 SARS-CoV-2 RNA，但人们对这些方案的性能知之甚少。在这项研究中，我们比较了从 35 个废水样本中检测 SARS-CoV-2 RNA 的三种基于反转录实时 PCR (RT-PCR) 的方案和一种基于液滴数字 PCR (ddPCR) 的方案。总体而言，85.7%的样本至少用一种方法检测出了 SARS-CoV-2 RNA，而用两种、三种或所有四种方法检测出 SARS-CoV-2 RNA 呈阳性的样本分别占 51.4%、22.8% 和 8.6%。基于商业 RT-PCR 检测法和液滴数字 PCR 的方案与内部检测法相比，总体灵敏度更高。针对不同基因使用一种以上的系统有助于提高检测灵敏度。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.

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