Loop-mediated isothermal amplification for rapid detection of Newcastle disease virus
Loop-mediated isothermal amplification for rapid detection of Newcastle disease virus
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DOI：
10.1128/jcm.43.4.1646-1650.2005
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发表时间：
2005-04-01
影响因子：
9.4
通讯作者：
Onuma, M
中科院分区：
文献类型：
--
作者：
Pham, HM;Nakajima, C;Onuma, M
我们评估了一种基于环介导的等温扩增(LAMP)分析的诊断系统，该系统可以直接从培养分离株和临床样本中快速、简单和灵敏地检测新城疫病毒(NDV)。通过使用一组针对融合蛋白基因的特异引物，LAMP方法在2小时内快速扩增出目标基因，只需常规的实验室水浴或热块即可进行反应。对38株新城疫病毒、其他不同毒株的基因组和实验感染鸡的临床样本的检测结果表明，LAMP与巢式PCR一样敏感和特异。所有LAMP阳性标本套式聚合酶链式反应均为阳性。LAMP检测比套式聚合酶链式反应更快、更经济、更易于操作。我们的结果清楚地表明，基于LAMP的检测方法是快速而敏感地诊断新城疫病毒感染的有用工具。
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.