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

Autor: Zhang Hao, Yang Fayu, Yang Man, Liu Jing, Wang Mi, Fei Chenzhong, Zhang Lifang, Xue Feiqun, Zhu Chuangang, Liu Yingchun, Gu Feng
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Acta Biochimica et Biophysica Sinica, Vol 54, Pp 1630-1636 (2022)
Druh dokumentu: article
ISSN: 1672-9145
DOI: 10.3724/abbs.2022163
Popis: 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.
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