Title Development of a rapid, sensitive detection method for SARS-CoV-2 and influenza virus based on recombinase polymerase amplification combined with CRISPR-Cas12a assay.
Author Wang, Yuning; Wu, Liqiang; Yu, Xiaomei; Wang, Gang; Pan, Ting; Huang, Zhao; Cui, Ting; Huang, Tianxun; Huang, Zhentao; Nie, Libo; Qian, Chungen
Journal J Med Virol Publication Year/Month 2023-Nov
PMID 37933907 PMCID -N/A-
Affiliation + expend 1.Department of Reagent Research and Development, Shenzhen YHLO Biotech Co., Ltd., Shenzhen, Guangdong, China.

Respiratory tract infections are associated with the most common diseases transmitted among people and remain a huge threat to global public health. Rapid and sensitive diagnosis of causative agents is critical for timely treatment and disease control. Here, we developed a novel method based on recombinase polymerase amplification (RPA) combined with CRISPR-Cas12a to detect three viral pathogens, including SARS-CoV-2, influenza A, and influenza B, which cause similar symptom complexes of flu cold in the respiratory tract. The detection method can be completed within 1 h, which is faster than other standard detection methods, and the limit of detection is approximately 10(2) copies/muL. Additionally, this detection system is highly specific and there is no cross-reactivity with other common respiratory tract pathogens. Based on this assay, we further developed a more simplified RPA/CRISPR-Cas12a system combined with lateral flow assay on a manual microfluidic chip, which can simultaneously detect these three viruses. This low-cost detection system is rapid and sensitive, which could be applied in the field and resource-limited areas without bulky and expensive instruments, providing powerful tools for the point-of-care diagnostic.

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    National Institute of Pathogen Biology, CAMS & PUMC, Bejing, China
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