Research on detection method of <i>Vibrio parahaemolyticus</i> based on CRISPR/Cas protein

Qun Wang; Zhe Li; Lin Zhao; Zijian Wang; Hongqun Zhao; Biao Kan; Bo Pang

doi:10.3784/j.issn.1003-9961.2020.06.012

Qun Wang, Zhe Li, Lin Zhao, Zijian Wang, Hongqun Zhao, Biao Kan, Bo Pang. Research on detection method of Vibrio parahaemolyticus based on CRISPR/Cas protein[J]. Disease Surveillance, 2020, 35(6): 513-517. DOI: 10.3784/j.issn.1003-9961.2020.06.012

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Research on detection method of Vibrio parahaemolyticus based on CRISPR/Cas protein

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ObjectiveTo establish a method for rapid detection of Vibrio parahaemolyticus by use the principle of clustered regularly interspaced short palindromic repeats (CRISPR) and the characteristics of Cas12a enzyme to achieve accurate and rapid detection and identification of pathogens.
MethodsIn this study, we prepared and purified Cas12a protein, screened and constructed V. parahaemolyticus guide RNA, established the CRISPR fluorescence detection system of V. parahaemolyticus, and determined the effectiveness of the CRISPR-VP detection method based on the final fluorescence amplification curve.
ResultsIn the CRISPR-VP detection method, a significant fluorescent signal will be produced only in the presence of the sequence of V. parahaemolyticus.
ConclusionThis experiment initially established the detection method of V. parahaemolyticus based on CRISPR/Cas protein, providing the preliminary conditions for the subsequent development of simple detection reagents.

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Research on detection method of Vibrio parahaemolyticus based on CRISPR/Cas protein

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