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
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, China.
Front Microbiol. 2022 Jun 9;13:920801. doi: 10.3389/fmicb.2022.920801. eCollection 2022.
Porcine epidemic diarrhoea virus (PEDV) is a member of the genus in the family . 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.
猪流行性腹泻病毒（PEDV）是科属的成员。它会导致仔猪急性水样腹泻和呕吐，死亡率很高。目前，GII基因型PEDV在野生毒株中的分离率很高，并且通常在免疫失败病例中被报道，这表明需要一种便携式且灵敏的检测方法。在此，逆转录重组酶辅助扩增（RT-RAA）与成簇规律间隔短回文重复序列（CRISPR）/Cas12a系统相结合，建立了一种用于PEDV的可多重化、快速且便携式的检测平台。针对GII型PEDV刺突（S）基因的CRISPR RNA（crRNA）被专门添加到该方案中。该系统适用于不同的实验条件，包括超灵敏荧光、可视化、紫外线或流式试纸检测。此外，它具有高灵敏度和特异性，每次反应中至少能检测到100个拷贝的靶基因。CRISPR/Cas12a检测平台所需时间较少，是快速诊断GII基因型PEDV的一种快速、可靠且实用的工具。
