Swarm primer-applied loop-mediated isothermal amplification (sLAMP) assay was developed for the rapid and specific detection of the ORF V1 gene of beak and feather disease virus (BFDV). The amplification can be completed in 40 min at 62 °C, and the results can be visually detected by the naked eye. The assay specifically amplified BFDV DNA and not amplified other viral nucleic acids. The limit of detection of the assay was 5 × 102 DNA copies/reaction, which was lower than that of the previously described LAMP (preLAMP) assay and comparable to that of a previously reported real time quantitative PCR (qPCR) assay. The detection rates of BFDV from psittacine clinical samples by the sLAMP, qPCR and preLAMP assays were 36.0 %, 36.0 % and 25.6 %, respectively, and the sLAMP results showed 100.0 % concordance with the qPCR results with a kappa value of 1.0. On the other hand, the preLAMP assay did not detect nine out of the 31 samples that were positive by sLAMP and qPCR assays, probably due to low sensitivity of the assay. These data suggest that the newly developed sLAMP assay will be a valuable tool for the rapid, sensitive, specific and reliable detection of BFDV in suspected psittacine birds, even in resource-limited laboratories.

中文翻译：

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一种先进的环介导的等温扩增测定法，用于快速检测鹦鹉鸟的喙和羽毛疾病病毒。

Swarm引物应用的环介导的等温扩增（sLAMP）分析法被开发出来，用于快速，特异性地检测喙和羽毛病病毒（BFDV）的ORF V1基因。扩增可以在62°C下40分钟内完成，并且可以用肉眼目测检测结果。该测定法特异性扩增了BFDV DNA，而不扩增其他病毒核酸。该测定的检测极限是5×102 DNA复制/反应，低于先前描述的LAMP（preLAMP）测定，与先前报道的实时定量PCR（qPCR）测定相当。sLAMP，qPCR和preLAMP分析检测的鹦鹉热临床样品中BFDV的检出率分别为36.0％，36.0％和25.6％，并且sLAMP结果与qPCR结果的一致性为100.0％，kappa值为1。0.另一方面，preLAMP检测未检测到sLAMP和qPCR检测呈阳性的31个样本中的9个，可能是由于该检测的敏感性较低。这些数据表明，即使在资源有限的实验室中，新开发的sLAMP测定法对于快速，灵敏，特异和可靠的可疑鹦鹉螺鸟类BFDV检测也将是有价值的工具。