Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin 150030, China.
J Virol Methods. 2012 Oct;185(1):18-23. doi: 10.1016/j.jviromet.2012.05.016. Epub 2012 May 29.
The objective of this study was to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of type II porcine reproductive and respiratory syndrome virus (PRRSV). Based on sequence alignment, four primers were designed amplifying the M gene of type II PRRSV and were subsequently utilized in an RT-LAMP assay. The RT-LAMP product had a ladder-like pattern of bands and the optimal reaction condition for this assay was determined to be 40 min at 63°C. Comparative analysis indicated that the RT-LAMP method was more sensitive than a conventional RT-PCR assay and comparable to a real-time PCR assay. In addition, the RT-LAMP assay was capable of detecting type II PRRSV in field samples and differentiating type II PRRSV from seven other porcine viruses which are all associated frequently with similar clinical symptoms.
本研究旨在开发一种逆转录环介导等温扩增（RT-LAMP）检测方法，用于检测 II 型猪繁殖与呼吸综合征病毒（PRRSV）。基于序列比对，设计了 4 个引物来扩增 II 型 PRRSV 的 M 基因，并随后将其用于 RT-LAMP 检测。RT-LAMP 产物具有梯状条带模式，最佳反应条件确定为 63°C 40 分钟。比较分析表明，该 RT-LAMP 方法比常规 RT-PCR 检测更灵敏，与实时 PCR 检测相当。此外，该 RT-LAMP 检测方法能够检测田间样本中的 II 型 PRRSV，并将其与七种其他常与类似临床症状相关的猪病毒区分开来。
