Surveillance of SARS-CoV-2 in wastewater by quantitative PCR and digital PCR: a case study in Shijiazhuang city, Hebei province, China.

Abstract

In this study, we reported the first long-term monitoring of SARS-CoV-2 in wastewater in Mainland China from November 2021 to October 2023. The city of Shijiazhuang was employed for this case study. We developed a triple reverse transcription droplet digital PCR (RT-ddPCR) method using triple primer-probes for simultaneous detection of the N1 gene, E gene, and Pepper mild mottle virus (PMMoV) to achieve accurate quantification of SARS-CoV-2 RNA in wastewater. Both the RT-ddPCR method and the commercial multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) method were implemented for the detection of SARS-CoV-2 in wastewater in Shijiazhuang City over a 24-month period. Results showed that SARS-CoV-2 was detected for the first time in the wastewater of Shijiazhuang City on 10 November 2022. The peak of COVID-19 cases occurred in the middle of December 2022, when the concentration of SARS-CoV-2 in the wastewater was highest. The trend of virus concentration increases and decreases forming a "long-tailed" shape in the COVID-19  outbreak and recession cycle. The results indicated that both multiplex RT-ddPCR and RT-qPCR are effective in detecting SARS-CoV-2 in wastewater, but RT-ddPCR is capable of detecting low concentrations of SARS-CoV-2 in wastewater which is more efficient. The SARS-CoV-2 abundance in wastewater is correlated to clinical data, outlining the public health utility of this work.HighlightsFirst long-term monitoring of SARS-CoV-2 in wastewater in Mainland ChinaCOVID-19 outbreak was tracked in Shijiazhuang City from outbreak to containmentWastewater was monitored simultaneously using RT-ddPCR and RT-qPCR methodsTriple primer-probe RT-ddPCR detects N1 and E genes of SARS-CoV-2 and PMMoV.

• Authors: Xiaoru Chai, Shiyou Liu, Chao Liu, Jiaxuan Bai, Juntao Meng, Hong Tian, Xu Han, Guangyue Han, Xiangdong Xu, Qi Li
• Journal: Emerging microbes & infections (Emerg Microbes Infect) Vol. 13 Issue 1 Pg. 2324502 (Dec 2024) ISSN: 2222-1751 [Electronic] United States
• PMID: 38465692 (Publication Type: Journal Article)

Chemical References

• Wastewater
• RNA, Viral

Topics

• Humans
• SARS-CoV-2 (genetics)
• Wastewater
• COVID-19 (diagnosis, epidemiology)
• RNA, Viral (genetics)
• China (epidemiology)
• Multiplex Polymerase Chain Reaction
• COVID-19 Testing
• Tobamovirus