Sensitive and specific method for rapid identification of Streptococcus pneumoniae using real-time fluorescence PCR
Sensitive and specific method for rapid identification of Streptococcus pneumoniae using real-time fluorescence PCR
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DOI：
10.1128/jcm.39.10.3446-3451.2001
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发表时间：
2001-10-01
影响因子：
9.4
通讯作者：
Lohman, KL
中科院分区：
文献类型：
--
作者：
McAvin, JC;Reilly, PA;Lohman, KL
病原体的分子监测表明，需要快速和可靠的方法来鉴定具有临床和流行病学重要性的生物。作为社区获得性肺炎的主要病原菌，本研究以肺炎链球菌为模式生物，建立和完善了实时荧光聚合酶链式反应和DNA提纯的改进方法。对经乳胶凝集试验证实的70株肺炎链球菌临床分离株与26株阴性对照临床分离株进行了热循环TaqMan法筛选。该探针由lytA基因构建，正确地检测了所有肺炎链球菌的基因组，没有与阴性对照发生交叉反应。这种方法的速度和案例将使其适用于许多细菌病原体的鉴定，并为适应从患者标本中直接检测提供了可能性。
Molecular surveillance of pathogens has shown the need for rapid and dependable methods for the identification of organisms of clinical and epidemiological importance. As the leading cause of community-acquired pneumonia, Streptococcus pneumoniae was used as a model organism to develop and refine a real-time fluorescence PCR assay and enhanced DNA purification method. Seventy clinical isolates of S. pneumoniae, verified by latex agglutination, were screened against 26 negative control clinical isolates employing a TaqMan assay on a thermocycler (LightCycler). The probe, constructed from the lytA gene, correctly detected all S. pneumoniae genomes without cross-reaction to negative controls. The speed and case of this approach will make it adaptable to identification of many bacterial pathogens and provide potential for adaptation to direct detection from patient specimens.