Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, China.
Beijing Scientific Observation and Experimental Station of Veterinary Drugs and Diagnostic Technology, Ministry of Agriculture, Beijing, 100193, China.
Arch Virol. 2021 Jun;166(6):1599-1605. doi: 10.1007/s00705-021-04963-w. Epub 2021 Mar 23.
Pigeon paramyxovirus-1 (PPMV-1) is a strain of Newcastle disease virus (NDV) that has adapted to infect pigeons and poses a constant threat to the commercial poultry industry. Early detection via rapid and sensitive methods, along with timely preventative and mitigating actions, is important for reducing the spread of PPMV-1. Here, we report the development of a TaqMan loop-mediated isothermal amplification assay (TaqMan-LAMP) for rapid and specific detection of PPMV-1 based on the F gene. This system makes use of six novel primers and a TaqMan probe that targets nine distinct regions of the F gene that are highly conserved among PPMV-1 isolates. The results showed that the limit of detection was 10 copies μL for PPMV-1 cDNA and 0.1 ng for PPMV-1 RNA. The reaction was completed within 25 min and was thus faster than conventional RT-PCR. Moreover, no cross-reactions with similar viruses or with peste des petits ruminants virus (PPRV) or NDV LaSota vaccine strains were observed under the same conditions. To evaluate the applicability of the assay, the TaqMan-LAMP assay and a commercial RT-PCR assay were compared using 108 clinical samples, and the concordance rate between two methods was found to be 96.3%. The newly developed PPMV-1 TaqMan-LAMP assay can therefore be used for simple, efficient, rapid, specific, and sensitive diagnosis of PPMV-1 infections.
鸽副黏病毒 1 型（PPMV-1）是一种适应感染鸽子的新城疫病毒（NDV）株，对商业家禽业构成持续威胁。通过快速和敏感的方法进行早期检测，并及时采取预防和缓解措施，对于减少 PPMV-1 的传播非常重要。在这里，我们报告了一种基于 F 基因的 TaqMan 环介导等温扩增检测（TaqMan-LAMP）方法的开发，用于快速特异性检测 PPMV-1。该系统利用六个新的引物和一个 TaqMan 探针，针对 F 基因中高度保守的九个不同区域进行靶标。结果表明，PPMV-1 cDNA 的检测限为 10 拷贝μL，PPMV-1 RNA 的检测限为 0.1ng。反应在 25 分钟内完成，因此比传统的 RT-PCR 更快。此外，在相同条件下，该反应与类似病毒或小反刍动物瘟病毒（PPRV）或 NDV LaSota 疫苗株没有交叉反应。为了评估该检测方法的适用性，使用 108 份临床样本比较了 TaqMan-LAMP 检测方法和商业 RT-PCR 检测方法，两种方法的一致性率为 96.3%。因此，新开发的 PPMV-1 TaqMan-LAMP 检测方法可用于简单、高效、快速、特异性和敏感的 PPMV-1 感染诊断。
