Rapid and visual detection of the emerging novel duck reovirus by using a specific and sensitive reverse transcription recombinase polymerase amplification method

Duck spleen necrosis disease (DSND) caused by Novel Reovirus (NDRV), is an emerging infectious that causes severely threaten to duck industry. Currently, the popular conventional RT-PCR technique for detecting NDRV time consuming. So, it essential develop a rapid and accurate molecular diagnosis techniques of pathogen purpose effective control disease. In our study, simple, reliable detection method was developed isothermal reverse transcription-recombinase polymerase amplification (RT-RPA). The RT-RPA primers were designed targeting S3 gene NDRV, its specificity verified testing series other waterfowl pathogens. A total 20 field experimental samples from infected ducklings tested compared with results quantitative simultaneously. could detect as little 4.14 × 102 copies/μl target in sensitivity analysis, which 10×higher sensitive than RT-PCR. major advantage be performed reaction at 37 ℃ completed within min. addition, no cross-reactivity detected waterfowl-origin viruses. Also, amplified products visualized faster, without gel electrophoresis, adding SYBR Green I observing them under ultraviolet light. newly offers especially useful on-site facilities resource-limited areas.