Establishment of DETECTR detection system for white spot syndrome virus
To establish a rapid,sensitive and specific DNA endonuclease targeted CRISPR trans re-porter(DETECTR)detection system for white spot syndrome virus(WSSV)by combining recombinase-medi-ated amplification(RAA)with CRISPR-Cas12a.RAA primers and crRNA were designed for the conserved genes of WSSV,and the optimal combination and best reaction condition were screened experimentally to con-struct the WSSV DETECTR detection system.The sensitivity and specificity of the system were tested and compared with the real-time PCR and Nested PCR methods recommended by WOAH.The Results showed that the WSSV DETECTR system has a minimum detection limit of 10 copies/μL,which is the same as real-time PCR and Nested PCR(3.34×101 copies/μL),there was no cross-reactivity with other pathogens and no differ-ence in the test results from real-time PCR and Nested PCR for clinical samples.In conclusion,WSSV DE-TECTR detection system has the advantages of being rapid and simple,sensitive and specific,and pro-vides a new method for the rapid detection of white spot syndrome virus.
recombinase-mediated amplificationCRISPR-Cas12awhite sport syndrome virusrapid detection
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