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LAMP assay coupled with CRISPR/Cas12a system for portable detection of African swine fever virus
  • +9
  • Bo YANG,
  • zhengwang shi,
  • Yuan Ma,
  • Lijuan Wang,
  • Liyan Cao,
  • Juncong Luo,
  • Ying Wang,
  • Rui Song,
  • Yiyong yan,
  • kehu yuan,
  • Hong Tian,
  • Haixue Zheng
Bo YANG
Lanzhou Veterinary Research Institute
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zhengwang shi
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Lijuan Wang
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Juncong Luo
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Yiyong yan
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Haixue Zheng
Lanzhou Veterinary Research Institute
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Abstract

African swine fever (ASF) is one of the most severe infectious diseases of pigs. In this study, a LAMP assay coupled with the CRISPR Cas12a system was established in one tube for the detection of the ASFV p72 gene. The single-strand DNA-fluorophore-quencher (ssDNA-FQ) reporters and CRISPR-derived RNA (crRNAs) were screened and selected for the CRISPR detection system. In combination with LAMP amplification assay, the detection limit for the LAMP-CRISPR assay can reach 7 copies/μl of p72 gene per reaction. Furthermore, this method displays no cross-reactivity with other porcine DNA or RNA viruses. The performance of the LAMP-CRISPR assay was compared with real-time qPCR tests for clinical samples, a good consistency between the LAMP-CRISPR assay and real-time qPCR was observed. In the current study, a LAMP coupled with the CRISPR detection method was developed. The method shed a light on the convenient, portable, low cost, highly sensitive and specific detection of ASFV, demonstrating a great application potential for monitoring on-site ASFV in the field.

Peer review status:IN REVISION

05 Feb 2021Submitted to Transboundary and Emerging Diseases
05 Feb 2021Assigned to Editor
05 Feb 2021Submission Checks Completed
10 Feb 2021Reviewer(s) Assigned
23 Mar 2021Review(s) Completed, Editorial Evaluation Pending
23 Mar 2021Editorial Decision: Revise Major