Analytical performance comparison of four SARS-CoV-2 RT-qPCR primer-probe sets for wastewater samples.
Sci Total Environ
; 806(Pt 2): 150572, 2022 Feb 01.
Article
in English
| MEDLINE | ID: covidwho-1433810
ABSTRACT
Current studies have confirmed the feasibility of SARS-CoV-2 RNA detection by RT-qPCR assays in wastewater samples as an effective surveillance tool of COVID-19 prevalence in a community. Analytical performance of various RT-qPCR assays has been compared against wastewater samples based on the positive ratio. However, there is no systematic comparison work has been conducted for both analytical sensitivity and quantitative reliability against wastewater, which are essential factors for WBE. In this study, the detection performance of four RT-qPCR primer-probe sets, including CCDC-N, CDC-N1, N-Sarbeco, and E-Sarbeco, was systematically evaluated with pure synthetized plasmids, spiked wastewater mocks and raw wastewater samples. In addition to confirm RT-qPCR results, Nanopore sequencing was employed to delineate at molecular level for the analytical sensitivity and reproducibility of those primer-probe sets. CCDC-N showed high sensitivity and the broadest linearity range for wastewater samples. It was thus recommended to be the most efficient tool in the quantitative analysis of SARS-CoV-2 in wastewater. CDC-N1 had the highest sensitivity for real wastewater and thus would be suitable for the screening of wastewater for the presence of SARS-CoV-2. When applying the primer-probe sets to wastewater samples collected from different Australian catchments, increased active clinical cases were observed with the augment of SARS-CoV-2 RNA quantified by RT-qPCR in wastewater in low prevalence communities.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
SARS-CoV-2
/
COVID-19
Type of study:
Diagnostic study
/
Experimental Studies
/
Observational study
/
Prognostic study
/
Systematic review/Meta Analysis
Limits:
Humans
Country/Region as subject:
Oceania
Language:
English
Journal:
Sci Total Environ
Year:
2022
Document Type:
Article
Affiliation country:
J.scitotenv.2021.150572
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