Zheng Su-Ya, Ma Li-Li, Wang Xiao-Li, Lu Li-Xin, Ma Sun-Ting, Xu Bin, Ouyang Wei
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China.
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Nanjing 210014, China.
J Virol Methods. 2022 Jun;304:114523. doi: 10.1016/j.jviromet.2022.114523. Epub 2022 Mar 12.
Infectious bursal disease (IBD), a major disease of birds, is caused by infectious bursal disease virus (IBDV). The disease can lead to immunosuppression, resulting in huge economic losses in the poultry industry. A specific, rapid, and simple detection method is important for the early diagnosis and prevention and control of IBDV. In this study, we established a naked-eye visual IBDV detection method, named "RPA-Cas12aDS", by combining recombinase polymerase amplification (RPA) with CRISPR-Cas12a-based nucleic acid detection. The detection process can be accomplished in 50 min, and uncapping contamination can be avoided. The detection results can be observed under blue or UV light. We used the RPA-Cas12aDS method to detect IBDV in bursa of Fabricius tissue samples of chickens, and the results were consistent with those obtained using commercial RT-PCR kits. This method presents great potential for visual, rapid, and point-of-care molecular diagnostics of IBDV in poultry.
传染性法氏囊病（IBD）是禽类的一种主要疾病，由传染性法氏囊病病毒（IBDV）引起。该疾病可导致免疫抑制，给家禽业造成巨大经济损失。一种特异性强、快速且简便的检测方法对于IBDV的早期诊断及防控至关重要。在本研究中，我们通过将重组酶聚合酶扩增（RPA）与基于CRISPR-Cas12a的核酸检测相结合，建立了一种名为“RPA-Cas12aDS”的肉眼可视IBDV检测方法。检测过程可在50分钟内完成，且可避免开盖污染。检测结果可在蓝光或紫外光下观察到。我们使用RPA-Cas12aDS方法检测鸡法氏囊组织样本中的IBDV，结果与使用商业RT-PCR试剂盒获得的结果一致。该方法在禽类IBDV的可视、快速及即时分子诊断方面具有巨大潜力。
