AUTHOR=Qian Bingxu , Liao Kai , Zeng Dexin , Peng Wanqing , Wu Xiaodong , Li Jinming , Bo Zongyi , Hu Yongxin , Nan Wenlong , Wen Yuan , Cao Yuying , Xue Feng , Zhang Xiaorong , Dai Jianjun 

TITLE=Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis

JOURNAL=Frontiers in Microbiology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2022.920801

DOI=10.3389/fmicb.2022.920801

ISSN=1664-302X

ABSTRACT=<p>Porcine epidemic diarrhoea virus (PEDV) is a member of the genus <italic>Alphacoronavirus</italic> in the family <italic>Coronaviridae</italic>. It causes acute watery diarrhoea and vomiting in piglets with high a mortality rate. Currently, the GII genotype, PEDV, possesses a high separation rate in wild strains and is usually reported in immunity failure cases, which indicates a need for a portable and sensitive detection method. Here, reverse transcription–recombinase aided amplification (RT-RAA) was combined with the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas12a system to establish a multiplexable, rapid and portable detection platform for PEDV. The CRISPR RNA (crRNA) against Spike (S) gene of GII PEDV specifically were added into the protocol. This system is suitable for different experimental conditions, including ultra-sensitive fluorescence, visual, UV light, or flow strip detection. Moreover, it exhibits high sensitivity and specificity and can detect at least 100 copies of the target gene in each reaction. The CRISPR/Cas12a detection platform requires less time and represents a rapid, reliable and practical tool for the rapid diagnosis of GII genotype PEDV.</p>