Highly Sensitive CRISPR/Cas12a-Based Fluorescence Detection of Porcine Reproductive and Respiratory Syndrome Virus

文献类型： 外文期刊

第一作者： Liu, Siyuan

作者： Liu, Siyuan;Chen, Bin;Liu, Siyuan;Yang, Yalan;Zhao, Yulan;Yang, Peng;Tang, Yijie;Tang, Zhonglin;Liu, Siyuan;Yang, Yalan;Zhao, Yulan;Yang, Peng;Tang, Yijie;Tang, Zhonglin;Tao, Dagang;Xie, Shengsong;Tao, Dagang;Xie, Shengsong;Liao, Yuying;Sun, Shouzhang;Liu, Yonggang;Yang, Peng;Tang, Zhonglin;Tang, Zhonglin;Tang, Zhonglin

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关键词： PRRSV; CRISPR-Cas12a; recombinase polymerase amplification; visualization; detection

期刊名称：ACS SYNTHETIC BIOLOGY （ 影响因子：5.11； 五年影响因子：5.239 ）

ISSN： 2161-5063

年卷期： 2021 年 10 卷 10 期

页码：

收录情况： SCI

摘要： Porcine reproductive and respiratory syndrome (PRRS) is an economically important disease of swine that is caused by PRRS virus (PRRSV). In this study, we established a fluorescence assay for highly sensitive detection of PRRSV through integration of the reverse transcription-recombinase polymerase amplification (RT-RPA)-coupled Cas12a system with an optical property of single stranded DNA-fluorescently quenched (ssDNA-FQ) reporter. This technique can achieve isothermal and visual detection of PRRSV in 25 min. In particular, the assay reaction can be completed in a single tube. The limit of sensitivity for PRRSV detection was single copy without cross-reactivity of other porcine viruses. Correlation between 11 PRRSV clinical samples measured by the quantitative reverse transcription polymerase chain reaction (RT-qPCR) and CRISPR/Cas12a assay was determined; the result showed that our results were highly accurate. To sum up, this study developed a visual, sensitive, and specific method of nucleic acid detection based on a CRISPR-Cas12a technique for the on-site detection of PRRSV.

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