Sensitive, specific, and rapid on-site detection of calf diarrhea pathogens using the RPA-CRISPR/Cas 12a assay
Yan Wang, Yun Diao, Tianqi Zhang, Fan Zhang, Wei Wang
TL;DR
A new rapid and sensitive test using RPA-CRISPR/Cas12a can detect four calf diarrhea pathogens on-site in under 50 minutes.
Contribution
A novel RPA-CRISPR/Cas12a assay for on-site detection of multiple calf diarrhea pathogens with high sensitivity and specificity.
Findings
The RPA-CRISPR/Cas12a assay detects BVDV, BCoV, BRV, and ETEC at concentrations as low as 10 copies/μL.
The assay is specific and does not produce false positives from other pathogens.
The test is completed in under 50 minutes in a single test tube without intermediate steps.
Abstract
Calf diarrhea is a common gastrointestinal disease that usually occurs within one month of birth. The disease causes the greatest economic losses to the cattle industry. Currently, a variety of diagnostic methods have been developed for calf diarrhea infections. However, existing methods are still unsatisfactory in terms of sensitivity, specificity, simplicity, cost, and speed.To provide a more sensitive, specific, simpler, and faster detection method, we recently developed an RPA-CRISPR/Cas12a assay that can detect BVDV, BCoV, BRV, and ETEC infections in cattle on-site. Testing for each pathogen is performed in a single test tube, without the need to open the tube in the middle, and can be completed in under 50 minutes.The RPA-CRISPR/Cas12a assay can detect BVDV, BCoV, BRV, and ETEC at concentrations of at least 10 copies/μL. The RPA-CRISPR/Cas12a assay does not produce false-positive…
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Taxonomy
TopicsAnimal Disease Management and Epidemiology · Clostridium difficile and Clostridium perfringens research · CRISPR and Genetic Engineering