Ultrasensitive and visual detection of pseudorabies virus based on CRISPR-Cas12b system

被引：2

作者：

Kang, Haoran ^{[1
]}

Yang, Xintan ^{[1
]}

Jiang, Ruijiao ^{[1
]}

Gao, Peng ^{[1
]}

Zhang, Yongning ^{[1
]}

Zhou, Lei ^{[1
]}

Ge, Xinna ^{[1
]}

Han, Jun ^{[1
]}

Guo, Xin ^{[1
]}

Yang, Hanchun ^{[1
]}

机构：

[1] China Agr Univ, Coll Vet Med, State Key Lab Vet Publ Hlth & Safety, Key Lab Anim Epidemiol,Minist Agr & Rural Affairs, Beijing, Peoples R China

来源：

MICROBIAL PATHOGENESIS | 2025年 / 203卷

关键词：

CRISPR-Cas12b system; Pseudorabies virus; Recombinase aided amplification; Quantitative PCR; Rapid detection; Point-of-care testing; CRISPR; CHINA; VARIANT;

D O I：

10.1016/j.micpath.2025.107447

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Aujeszky's disease (AD) is an acute infectious disease that infects pigs and other animals, resulting in significant economic losses and posing a threat to human health. Reliable and rapid detection methods are essential for the prevention of AD. In this study, a RAA-Cas12b assay based on recombinase-aided amplification (RAA) and CRISPR-Cas12b system was established, optimized and evaluated for the rapid detection of wild-type Pseudorabies Virus (PRV). The results can not only be detected by real-time fluorescence readout, but also can be visualized by a portable blue light instrument. There was no cross-reaction with PRV Bartha-K61 strain or other swine infectious viruses. The analytical sensitivities of the real-time PRV RAA-Cas12b assay and visual PRV RAACas12b assay were determined to be 15 copies/mu L with 95 % confidence interval and 140 copies/mu L with 95 % confidence interval, respectively. A total of 31 clinical samples were detected and compared with PRV qPCR assay to evaluate the diagnostic performance of the PRV RAA-Cas12b assay. The diagnostic coincidence rate of the two assays was 100 %. In summary, this convenient and reliable assay has great potential for rapid detection of wild type PRV in point-of-care testing (POCT).

引用

页数：8

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