Rapid identification and strain-typing of respiratory pathogens for epidemic surveillance
Rapid identification and strain-typing of respiratory pathogens for epidemic surveillance
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DOI：
10.1073/pnas.0409920102
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发表时间：
2005-05-31
影响因子：
11.1
通讯作者：
Hofstadler, SA
中科院分区：
文献类型：
--
作者：
Ecker, DJ;Sampath, R;Hofstadler, SA
流行性呼吸道感染是造成军人和平民大量发病和死亡的原因。我们描述了一种高通量的方法，同时识别和基因型的呼吸道样本中的复杂混合物的细菌物种。该方法使用电喷雾电离质谱和来自高度保守基因组区域的PCR扩增产物的碱基组成分析来鉴定和确定样品中存在的病原菌的相对量。通过使用针对特定细菌基因组高度可变区域的额外引物来实现特定物种的高分辨率基因分型。这种方法被用来检查呼吸道疾病爆发期间从新兵中采集的样本，并在几个军事训练设施进行后续监测。对呼吸道样本的分析显示，包括流感嗜血杆菌、脑膜炎奈瑟菌和化脓性链球菌在内的呼吸道致病菌浓度很高。当S.从流行点的样本中鉴定出化脓性链球菌，在几乎所有新兵中发现了相同的基因型。这种分析方法将为了解呼吸道疾病爆发性流行的多微生物性质提供基本信息。
Epidemic respiratory infections are responsible for extensive morbidity and mortality within both military and civilian populations. We describe a high-throughput method to simultaneously identify and genotype species of bacteria from complex mixtures in respiratory samples. The process uses electrospray ionization mass spectrometry and base composition analysis of PCR amplification products from highly conserved genomic regions to identify and determine the relative quantity of pathogenic bacteria present in the sample. High-resolution genotyping of specific species is achieved by using additional primers targeted to highly variable regions of specific bacterial genomes. This method was used to examine samples taken from military recruits during respiratory disease outbreaks and for follow up surveillance at several military training facilities. Analysis of respiratory samples revealed high concentrations of pathogenic respiratory species, including Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pyogenes. When S. pyogenes was identified in samples from the epidemic site, the identical genotype was found in almost all recruits. This analysis method will provide information fundamental to understanding the polymicrobial nature of explosive epidemics of respiratory disease.