The Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang 150001, China.
J Virol Methods. 2011 Apr;173(1):31-6. doi: 10.1016/j.jviromet.2011.01.002. Epub 2011 Jan 8.
This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for distinguishing avian leukosis virus (ALV) subgroup A from other subgroups of the virus. On the basis of the results of sequence comparison and the sequence characteristics of ALV subgroups, a LAMP method was designed to target the gp85 segment for detection of ALV-A. Under optimal reaction conditions, ALV-A LAMP produced neither cross-reactions with other major subgroups (including subgroups J, B, C, and E) nor nonspecific reactions with other common avian infectious diseases. A sensitivity test showed that this method can detect 20 copies of proviral nucleic acid sequence within 45 min, which is 100 times more sensitive than the conventional polymerase chain reaction (PCR). This method can detect subgroup A virus rapidly and the results can be assessed based on color changes. The whole reaction process can be performed without opening the lid of the reaction tube, which reduces the possibility of contamination greatly and simplifies the detection process, indicating the considerable potential of this method for in situ application in the future.
本研究旨在建立一种环介导等温扩增（LAMP）方法，用于区分禽白血病病毒（ALV）亚群 A 与病毒的其他亚群。基于序列比较的结果和 ALV 亚群的序列特征，设计了一种针对 gp85 片段的 LAMP 方法，用于检测 ALV-A。在最佳反应条件下，ALV-A LAMP 既不与其他主要亚群（包括亚群 J、B、C 和 E）发生交叉反应，也不与其他常见的禽传染病发生非特异性反应。灵敏度测试表明，该方法可在 45 分钟内检测到 20 个拷贝的前病毒核酸序列，比常规聚合酶链反应（PCR）灵敏 100 倍。该方法可快速检测亚群 A 病毒，结果可通过颜色变化进行评估。整个反应过程无需打开反应管盖，可大大降低污染的可能性，简化检测过程，表明该方法在未来具有很大的现场应用潜力。
