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通过聚合酶链反应(PCR)结合电喷雾电离质谱法对腺病毒进行快速检测和分子血清分型。

Rapid detection and molecular serotyping of adenovirus by use of PCR followed by electrospray ionization mass spectrometry.

作者信息

Blyn Lawrence B, Hall Thomas A, Libby Brian, Ranken Raymond, Sampath Rangarajan, Rudnick Karl, Moradi Emily, Desai Anjali, Metzgar David, Russell Kevin L, Freed Nikki E, Balansay Melinda, Broderick Michael P, Osuna Miguel A, Hofstadler Steven A, Ecker David J

机构信息

Ibis Biosciences, Inc., Carlsbad, CA 92008, USA.

出版信息

J Clin Microbiol. 2008 Feb;46(2):644-51. doi: 10.1128/JCM.00801-07. Epub 2007 Dec 19.

Abstract

We have developed a PCR/electrospray ionization mass spectrometry (PCR/ESI-MS) assay for the rapid detection, identification, and serotyping of human adenoviruses. The assay employs a high-performance mass spectrometer to "weigh" the amplicons obtained from PCR using primers designed to amplify known human adenoviruses. Masses are converted to base compositions and, by comparison against a database of the genetic sequences, the serotype present in a sample is determined. The performance of the assay was demonstrated with quantified viral standards and environmental and human clinical samples collected from a military training facility. Over 500 samples per day can be analyzed with sensitivities greater than 100 genomes per reaction. This approach can be applied to many other families of infectious agents for rapid and sensitive analysis.

摘要

我们开发了一种用于快速检测、鉴定和血清分型人腺病毒的聚合酶链反应/电喷雾电离质谱法(PCR/ESI-MS)。该检测方法采用高性能质谱仪对使用设计用于扩增已知人腺病毒的引物通过PCR获得的扩增子进行“称重”。质量被转换为碱基组成,并通过与遗传序列数据库进行比较,确定样品中存在的血清型。使用定量病毒标准品以及从军事训练设施收集的环境和人类临床样本证明了该检测方法的性能。每天可分析超过500个样本,灵敏度大于每个反应100个基因组。这种方法可应用于许多其他感染因子家族,以进行快速和灵敏的分析。

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