AUTHOR=Ma Lei , Zhu Mengjie , Meng Qingfeng , Wang Yao , Wang Xueping 

TITLE=Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=Volume 13 - 2023

YEAR=2024

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2023.1305222

DOI=10.3389/fcimb.2023.1305222

ISSN=2235-2988

ABSTRACT=Senecavirus A (SVA) is a highly contagious virus that causes vesicular disease in pigs. At present, laboratory detection methods, such as virus isolation and polymerase chain reaction (PCR), required precision instruments and qualified personnel, making them unsuitable for point-of-care tests (POCT). Fortunately, the emergence of CRISPR/Cas system has provided new opportunities for fast and efficient pathogen detection. This study successfully developed a precise and sensitive detection platform for diagnosing SVA by combining the CRISPR system with recombinase polymerase amplification (RPA). The minimum detection limit of the assay was 10 copies of the SVA genome.Meanwhile, the assay demonstrated high specificity. To validate the effectiveness of this system, we tested 85 swine clinical samples and found that the fluorescence method had a 100% coincidence rate compared to RT-qPCR. Overall, the RPA-CRISPR/Cas12a assay established in our study is a highly effective method for detecting SVA and holds great potential for practical applications in the resource-limited settings.1. A real-time One-Tube RPA-CRISPR/Cas12a assay was established for the detection of Seneca Valley virus 2. The performance of the assay was as good as that of RT-qPCR 3. The assay was a powerful diagnostic platform that offers an alternative to detect SVA in marginalized areas