College of Veterinary Medicine, South China Agricultural University, No.483 Wushan Road, Tianhe District, Guangzhou, 510642, People's Republic of China.
Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, South China Agricultural University, Guangzhou, 510642, People's Republic of China.
Sci Rep. 2021 May 26;11(1):10946. doi: 10.1038/s41598-021-90479-x.
Avian leukosis virus subgroup J (ALV-J) causes oncogenic disease in chickens in China, resulting in great harm to poultry production, and remains widespread in China. Herein, we employed a cross-priming amplification (CPA) approach and a nucleic acid detection device to establish a visual rapid detection method for ALV-J. The sensitivity of CPA, polymerase chain reaction (PCR) and real-time PCR (RT-PCR) was compared, and the three methods were used to detect ALV-J in the cell cultures which inoculated with clinical plasma. The result showed when the amplification reaction was carried out at 60 °C for just 60 min, the sensitivity of CPA was 10 times higher than conventional PCR, with high specificity, which was comparable with RT-PCR, based on detection of 123 cell cultures which inoculated with clinical plasma, the coincidence rate with real-time PCR was 97.3% (71/73). CPA detection of ALV-J does not require an expensive PCR instrument; a simple water bath or incubator is sufficient for complete DNA amplification, and the closed nucleic acid detection device avoids aerosol pollution, making judgment of results more intuitive and objective. The CPA assay would be a promising simple, rapid and sensitive method for identification of ALV-J.
禽白血病病毒亚群 J（ALV-J）在中国引起鸡的致癌疾病，对家禽生产造成了极大的危害，并在中国广泛存在。在此，我们采用交叉引物扩增（CPA）方法和核酸检测装置，建立了一种用于快速检测 ALV-J 的可视化方法。比较了 CPA、聚合酶链反应（PCR）和实时 PCR（RT-PCR）的灵敏度，并使用这三种方法检测接种了临床血浆的细胞培养物中的 ALV-J。结果表明，当在 60°C 下仅进行 60 分钟的扩增反应时，CPA 的灵敏度比常规 PCR 高 10 倍，具有高特异性，与 RT-PCR 相当，基于对接种了临床血浆的 123 个细胞培养物的检测，与实时 PCR 的符合率为 97.3%（71/73）。CPA 检测 ALV-J 不需要昂贵的 PCR 仪器；简单的水浴或孵育箱足以完成 DNA 扩增，并且封闭的核酸检测装置可避免气溶胶污染，使结果判断更直观和客观。CPA 检测方法可能是一种有前途的简单、快速和敏感的 ALV-J 鉴定方法。
