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

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

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

Author(s) -

Siyuan Liu,

Dagang Tao,

Yuying Liao,

Yalan Yang,

Shouzhang Sun,

Yulan Zhao,

Peng Yang,

Yijie Tang,

Bin Chen,

Yonggang Liu,

Shengsong Xie,

Zhonglin Tang

Publication year - 2021

Publication title -

acs synthetic biology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.156

H-Index - 66

ISSN - 2161-5063

DOI - 10.1021/acssynbio.1c00103

Subject(s) - porcine reproductive and respiratory syndrome virus , recombinase polymerase amplification , crispr , biology , virology , reverse transcriptase , polymerase chain reaction , microbiology and biotechnology , detection limit , trans activating crrna , loop mediated isothermal amplification , nucleic acid , dna , virus , real time polymerase chain reaction , cas9 , gene , chemistry , genetics , chromatography

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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