Development of a DNA microarray to identify the Streptococcus pneumoniae serotypes contained in the 23-valent pneumococcal polysaccharide vaccine and closely related serotypes
Development of a DNA microarray to identify the Streptococcus pneumoniae serotypes contained in the 23-valent pneumococcal polysaccharide vaccine and closely related serotypes
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开发DNA微阵列来鉴定23价肺炎球菌多糖疫苗中所含的肺炎链球菌血清型以及密切相关的血清型
DOI：
10.1016/j.mimet.2006.07.001
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发表时间：
2007-01-01
影响因子：
2.2
通讯作者：
Feng, Lu
中科院分区：
文献类型：
--
作者：
Wang, Quan;Wang, Min;Feng, Lu
肺炎链球菌是一种世界性的主要人类病原体。本研究建立了一种可靠、准确的肺炎链球菌种间分化和23组肺炎链球菌种内分化的DNA微阵列方法，包括23价肺炎球菌疫苗所代表的血清型和其他20个密切相关的血清型。除了16S rDNA探针外，还产生了针对荚膜多糖合成(Cps)基因、wzy或capA的血清型或血清组特异性探针。我们采用两步多重聚合酶链式反应将检测灵敏度提高到纯培养或50 ng DNA中的10(5)cfu/ml。对来自中国、澳大利亚、加拿大和新西兰的169株肺炎链球菌进行了检测，包括23价疫苗和密切相关血清型的147株，其他血清型的11株，以及从呼吸道中分离的不同种类的11株。开发的DNA微阵列方法提供了一种敏感的手段，可以快速识别患者中最常见的肺炎链球菌血清型成员，并监测它们在不同患者群体和地理位置的分布。这些信息对于疾病监测和疫苗效力监测是必要的。(C)2006爱思唯尔B.V.保留所有权利。
Streptococcus pneumoniae is a major worldwide human pathogen. This investigation has developed a reliable and accurate DNA microarray method for inter-species differentiation of S. pneumoniae and intra-species differentiation of the 23 groups of S. pneumoniae including serotypes represented in the 23-valent pneumococcal vaccine and the other 20 closely related serotypes. In addition to 16S rDNA probes, serotype- or serogroup-specific probes targeting the capsular polysaccharide synthesis (cps) genes, wzy or capA were generated. We adopted a two-step multiplex PCR to improve the sensitivity of detection to a level of 10(5) cfu/ml in pure culture or 50 ng DNA. A total of 169 isolates (from China, Australia, Canada and New Zealand) including 147 belonging to 23-valent vaccine and closely related serotypes of S. pneumoniae, 11 belonging to other serotypes and 11 of different species commonly isolated from respiratory tract were tested to verify the method. The DNA microarray method developed provides a sensitive means to rapidly identify the members of the most common S. pneumoniae serotypes in patients and to monitor their distribution in different patient groups and geographic locations. Such information is needed for disease surveillance and to monitor vaccine efficacy. (c) 2006 Elsevier B.V. All rights reserved.