Virus Surveillance and Diagnosis Branch, Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA 30333, USA.
Expert Rev Mol Diagn. 2011 Jan;11(1):41-52. doi: 10.1586/erm.10.107.
Reverse-transcription PCR (RT-PCR) coupled with electrospray ionization mass spectrometry (ESI-MS) is a high-throughput nucleic acid-based technology that relies on the accurate measurement of the molecular weight of PCR amplicons that can be used to deduce the base counts (number of As, Gs, Cs and Ts) of DNA. These amplicons represent highly variable regions with information-rich sequences, which are flanked by broad-range primers designed based on highly conserved loci. This technology was first introduced in 2005 for microbial identification and subtyping, and was later applied to influenza virus detection and identification. The influenza RT-PCR/ESI-MS assay allows analysis of approximately 300 samples per 24 h, and aids in the characterization of influenza viruses based on their 'core' gene signatures. Notably, this assay was used to identify one of the first cases of the 2009 H1N1 pandemic viruses. One of the main advantages of the RT-PCR/ESI-MS technology is its universality and adaptability for pathogen characterization. Efforts are being made to customize the currently used influenza surveillance assay for use in the diagnosis of the H1N1 pandemic virus. In this article, we provide a summary of known applications of the RT-PCR/ESI-MS assay in the field of influenza.
逆转录聚合酶链反应（RT-PCR）结合电喷雾电离质谱（ESI-MS）是一种高通量的基于核酸的技术，它依赖于对 PCR 扩增子分子量的精确测量，可用于推断 DNA 的碱基计数（A、G、C 和 T 的数量）。这些扩增子代表具有丰富信息序列的高度可变区域，其侧翼是基于高度保守基因座设计的广谱引物。该技术于 2005 年首次用于微生物鉴定和亚型分析，后来也应用于流感病毒检测和鉴定。流感 RT-PCR/ESI-MS 检测法允许每 24 小时分析大约 300 个样本，并有助于根据流感病毒的“核心”基因特征对其进行特征描述。值得注意的是，该检测法用于鉴定 2009 年 H1N1 大流行病毒的首例病例之一。RT-PCR/ESI-MS 技术的主要优势之一是其通用性和适用于病原体特征描述的适应性。目前正在努力定制现有的流感监测检测法，以用于诊断 H1N1 大流行病毒。本文总结了 RT-PCR/ESI-MS 检测法在流感领域的已知应用。
