Pham Hang Minh, Nakajima Chie, Ohashi Kazuhiko, Onuma Misao
Laboratory of Infectious Diseases, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
J Clin Microbiol. 2005 Apr;43(4):1646-50. doi: 10.1128/JCM.43.4.1646-1650.2005.
We have evaluated a diagnostic system based on the loop-mediated isothermal amplification (LAMP) assay for the rapid, simple, and sensitive detection of Newcastle disease virus (NDV) directly from culture isolates as well as clinical samples. By using one set of specific primers targeting the fusion protein gene, the LAMP assay rapidly amplified the target gene within 2 h, requiring only a regular laboratory water bath or heat block for reaction. The results obtained from testing the genomes of 38 NDV strains, other different viruses, and clinical samples of experimentally infected chickens showed that LAMP was as sensitive and specific as nested PCR. All LAMP-positive samples were positive by nested PCR. The LAMP assay is faster than nested PCR, cost-effective, and easy to perform. Our results clearly demonstrate that the LAMP-based assay is a useful tool for the rapid and sensitive diagnosis of NDV infection.
我们评估了一种基于环介导等温扩增（LAMP）检测法的诊断系统，用于直接从培养分离株以及临床样本中快速、简便且灵敏地检测新城疫病毒（NDV）。通过使用一组靶向融合蛋白基因的特异性引物，LAMP检测法在2小时内快速扩增了目标基因，反应仅需常规实验室水浴或热块。对38株NDV毒株的基因组、其他不同病毒以及实验感染鸡的临床样本进行检测的结果表明，LAMP与巢式PCR一样灵敏且特异。所有LAMP阳性样本经巢式PCR检测均为阳性。LAMP检测法比巢式PCR更快、成本效益更高且易于操作。我们的结果清楚地表明，基于LAMP的检测法是快速灵敏诊断NDV感染的有用工具。
