Rapid and sensitive detection of Trichomonas gallinae using RAA-CRISPR-Cas12a.

Abstract

Trichomonas gallinae (T. gallinae) is an important pathogen causing trichomoniasis in birds, especially pigeons. Rapid and sensitive detection methods for T. gallinae are urgently needed to diagnose T. gallinae early to reduce poultry industry losses. Therefore, we developed a rapid and sensitive diagnostic method based on recombinase-aided amplification (RAA) assay and clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 12a (CRISPR/Cas12a) system to detect T. gallinae. The RAA-CRISPR/Cas12a method can be divided into RAA-CRISPR/Cas12a fluorescent signal (RAA-CRISPR/Cas12a-FL) and RAA-CRISPR/Cas12a lateral flow strip (RAA-CRISPR/Cas12-LFS). Both RAA-CRISPR/Cas12a-FL and RAA-CRISPR/Cas12-LFS methods show the property of rapid. sensitive, and does not require a sophisticated instrument, and they allow the detection of T. gallinae in less than 1 hr. Meanwhile, they have satisfactory specificity and can accurately detect T. gallinae in samples of different pathogens. In summary, the RAA-CRISPR/Cas12a-FL and RAA-CRISPR/Cas12-LFS methods we constructed can be used for on-site T. gallinae detection and resource-poor areas.