Development of a DNA Microarray Method for Detection and Identification of All 15 Distinct O-Antigen Forms of Legionella pneumophila
Development of a DNA Microarray Method for Detection and Identification of All 15 Distinct O-Antigen Forms of Legionella pneumophila
复制标题
开发用于检测和鉴定嗜肺军团菌所有 15 种不同 O 抗原形式的 DNA 微阵列方法
DOI：
10.1128/aem.01957-13
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发表时间：
2013-11-01
影响因子：
4.4
通讯作者：
Wang, Lei
中科院分区：
文献类型：
--
作者：
Cao, Boyang;Yao, Fangfang;Wang, Lei
摘要军团菌在许多环境中普遍存在。至少有50种和70个血清型的革兰氏阴性菌已被确定。在这50个菌种中，20个是致病性的，嗜肺军团菌是绝大多数（约90%）发生的军团菌病病例的原因。此外，在15 L。在已鉴定的嗜肺军团菌血清群中，仅O 1就引起了全球发生的军团病病例的84%以上。快速可靠的检测和鉴定L.水、环境和临床样品中的嗜肺菌的需求量很大。L.嗜肺细菌传统上通过免疫学方法通过它们的O抗原来鉴定。我们最近开发了一种O血清群特异性DNA微阵列，用于检测所有15种不同的O抗原形式的L。嗜肺菌，包括血清群O 1至O 15。共使用35个菌株验证微阵列的特异性，其中包括15个L。嗜肺军团菌O抗原标准参考菌株和7株L.嗜肺军团菌临床分离株作为靶菌株，其他非嗜肺军团菌属种的7种参考菌株作为密切相关菌株，6种非军团菌属细菌种作为非相关菌株。经滤膜富集和平板培养，检测灵敏度可达1 ng基因组DNA或0.4CFU/ml水样。本研究表明，微阵列可以特异性，灵敏度和重复性检测L。嗜肺菌血清群。据我们所知，这是第一个报告的微阵列血清分型方法的所有15个不同的O-抗原形式的L。嗜肺菌
ABSTRACT Legionella is ubiquitous in many environments. At least 50 species and 70 serogroups of the Gram-negative bacterium have been identified. Of the 50 species, 20 are pathogenic, and Legionella pneumophila is responsible for the great majority (approximately 90%) of the Legionnaires' disease cases that occur. Furthermore, of the 15 L. pneumophila serogroups identified, O1 alone causes more than 84% of the Legionnaires' disease cases that occur worldwide. Rapid and reliable assays for the detection and identification of L. pneumophila in water, environmental, and clinical samples are in great demand. L. pneumophila bacteria are traditionally identified by their O antigens by immunological methods. We have recently developed an O serogroup-specific DNA microarray for the detection of all 15 distinct O-antigen forms of L. pneumophila, including serogroups O1 to O15. A total of 35 strains were used to verify the specificity of the microarray, including 15 L. pneumophila O-antigen standard reference strains and seven L. pneumophila clinical isolates as target strains, seven reference strains of other non-pneumophila Legionella species as closely related strains, and six non-Legionella bacterial species as nonrelated strains. The detection sensitivity was 1 ng of genomic DNA or 0.4 CFU/ml in water samples with filter enrichment and plate culturing. This study demonstrated that the microarray allows specific, sensitive, and reproducible detection of L. pneumophila serogroups. To the best of our knowledge, this is the first report of a microarray serotyping method for all 15 distinct O-antigen forms of L. pneumophila.