An ultrasensitive, rapid and portable method for screening oseltamivir-resistant virus based on CRISPR/Cas12a combined with immunochromatographic strips

Zhang Hao; Yang Fayu; Yang Man; Liu Jing; Wang Mi; Fei Chenzhong; Zhang Lifang; Xue Feiqun; Zhu Chuangang; Liu Yingchun; Gu Feng

doi:10.3724/abbs.2022163

Acta Biochimica et Biophysica Sinica (Nov 2022)

An ultrasensitive, rapid and portable method for screening oseltamivir-resistant virus based on CRISPR/Cas12a combined with immunochromatographic strips

Zhang Hao,
Yang Fayu,
Yang Man,
Liu Jing,
Wang Mi,
Fei Chenzhong,
Zhang Lifang,
Xue Feiqun,
Zhu Chuangang,
Liu Yingchun,
Gu Feng

Affiliations

Zhang Hao: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Yang Fayu: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Yang Man: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","National Reference Laboratory for Animal Schistosomiasis, Shanghai 200241, China"]
Liu Jing: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Wang Mi: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Fei Chenzhong: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Zhang Lifang: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Xue Feiqun: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Zhu Chuangang: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","National Reference Laboratory for Animal Schistosomiasis, Shanghai 200241, China"]
Liu Yingchun: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]
Gu Feng: ["Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China","Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai 200241, China"]

DOI: https://doi.org/10.3724/abbs.2022163
Journal volume & issue: Vol. 54
pp. 1630 – 1636

Abstract

Read online

Influenza is a significant public health challenge because of the emergence of antigenically shifted or highly virulent strains. The neuraminidase inhibitor oseltamivir is used as an antiviral drug in clinical treatment. However, its therapeutic effects can be greatly compromised by the emergence of drug-resistant mutant viruses. Thus, there is an urgent need to distinguish drug-resistant strains with a simple method. To address this, in the present study, we develop a rapid, sensitive and convenient molecular diagnosis method based on CRISPR/Cas12a technology and lateral flow detection (LFD). By targeting mutant sequences amplified by recombinase polymerase amplification (RPA) reaction, crRNA is designed to develop the CRISPR/Cas12a assay, and 2000 copies can be directly observed by the naked eye under blue light-emitting diode (LED) light. Combined with LFD, the limit of detection of RPA-CRISPR/Cas12a-LFD is about 20 copies of target sequence per reaction. Collectively, RPA-CRISPR/Cas12a-LFD method provides a novel alternative for the sensitive, specific and portable detection to diagnose oseltamivir-resistant mutant strains.

Published in Acta Biochimica et Biophysica Sinica

ISSN: 1672-9145 (Print); 1745-7270 (Online)
Publisher: China Science Publishing & Media Ltd.
Country of publisher: China
LCC subjects: Science: Chemistry: Organic chemistry: Biochemistry; Science: Biology (General): Genetics
Website: https://www.sciengine.com/ABBS/home

About the journal

Abstract

Keywords