Structural comparison of O-antigen gene clusters of Legionella pneumophila and its application of a serogroup-specific multiplex PCR assay
Structural comparison of O-antigen gene clusters of Legionella pneumophila and its application of a serogroup-specific multiplex PCR assay
复制标题
嗜肺军团菌O抗原基因簇的结构比较及其血清群特异性多重PCR检测的应用
DOI：
10.1007/s10482-015-0594-0
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发表时间：
2015-12-01
影响因子：
2.6
通讯作者：
Wang, Lei
中科院分区：
文献类型：
--
作者：
Cao, Boyang;Tian, Zhenyang;Wang, Lei
肺炎军团血清群O1，O4，O6，O7，O10和O13是与肺炎相关的致病菌株。对肺炎乳杆菌的表面O-抗原基因簇O4，O6，O7，O10和O13进行了测序和分析，并基于与肺炎杆菌的基因相同的功能，并以同源性注释。六氯杆菌血清群的基因基因座含有YVFE，NeuaBCD，psea类的基因，用于核苷酸糖生物合成，WECA用于糖转移的WECA和WZM以及用于O-抗原处理的WZT。 O-抗原基因的检测允许在物种和血清群水平上进行细分分化，而无需核苷酸测序。 O抗原处理的基因WZM和WZT在不同的血清群中被发现是独特的，它已被用作检测和鉴定不同O血清群的肺炎杆菌菌株的靶基因。在本报告中，开发了基于WZM或WZT的多重PCR分析，该测定法与新设计的O1和O7的新设计的特定引物对开发了所有六个血清群，以及用于O4，O6，O6，O10和O13的特定引物对。通过对34个菌株的分析来验证该阵列，其中包括15个肺炎。检测敏感性是一种NG基因组DNA。准确和敏感的测定适用于环境和临床样品中这些血清群的菌株的鉴定和检测。
The Legionella pneumophila serogroups O1, O4, O6, O7, O10 and O13 are pathogenic strains associated with pneumonia. The surface O-antigen gene clusters of L. pneumophila serogroups O4, O6, O7, O10 and O13 were sequenced and analyzed, with the function annotated on the basis of homology to that of the genes of L. pneumophila serogroup O1 (L. pneumophila subsp. pneumophila str. Philadelphia 1). The gene locus of the six L. pneumophila serogroups contains genes of yvfE, neuABCD, pseA-like for nucleotide sugar biosynthesis, wecA for sugar transfer, and wzm as well as wzt for O-antigen processing. The detection of O-antigen genes allows the fine differentiation at species and serogroup level without the neccessity of nucleotide sequencing. The O-antigen-processing genes wzm and wzt, which were found to be distinctive for different for different serogroups, have been used as the target genes for the detection and identification of L. pneumophila strains of different O serogroups. In this report, a multiplex PCR assay based on wzm or wzt that diferentiates all the six serogroups by amplicon size was developed with the newly designed specific primer pairs for O1 and O7, and the specific primer pairs for O4, O6, O10, and O13 reported previously. The array was validated by analysis of 34 strains including 15 L. pneumophila O-standard reference strains, eight reference strains of other Legionella non-pneumophila species, six other bacterial species, and five L. pneumophila environmental isolates. The detection sensitivity was one ng genomic DNA. The accurate and sensitive assay is suitable for the identification and detection of strains of these serogroups in environmental and clinical samples.