Application of portable real-time recombinase-aided amplification (rt-RAA) assay in the clinical diagnosis of ASFV and prospective DIVA diagnosis.

摘要: African swine fever, a serious infectious disease, has been found in many countries around the world over last nearly 100 years, and causes untold damage to economy wherever it occurs. Diagnosis is currently performed by real-time PCR, which highly sensitive but can only be carried out diagnostic laboratory environment with sophisticated equipment expertise. A sensitive, rapid method that implemented agricultural settings thus urgently needed for detection control of fever virus (ASFV) infection. In this study, we developed an isothermal amplification technology achieve molecular diagnosis ASFV clinical samples, using recombinase-aided (RAA) assay combined portable instrument. This avoids limitations traditional PCR offers times within 20 min, enabling as few 10 copies DNA molecules per reaction without cross-reaction other common viruses. We evaluated performance 200 blood samples. The coincidence rate results between rt-RAA RT-qPCR was 96.94% positive, 100% negative, 97.50% total. have also system targeting EP402R gene, reaction; possibility DIVA (differentiating infected from vaccinated animals) diagnosis, because CD2V gene-deleted could soon approved leading candidate live attenuated vaccine China. reliable, rapid, method, reasonable alternative point-of-care ASFV. KEY POINTS: • RT-RAA detect genome min. different genes differentiating diagnosis.