Targeting single-nucleotide polymorphisms in the 16S rRNA gene to detect and differentiate Legionella pneumophila and non-Legionella pneumophila species

被引：3
作者：
Zhan, Xiao-Yong [1 ,2 ,3 ]
Hu, Chao-Hui [1 ,2 ]
Zhu, Qing-Yi [1 ,2 ]
机构：
[1] Guangzhou KingMed Ctr Clin Lab, 10,Luoxuan 3 Rd, Guangzhou 510300, Guangdong, Peoples R China
[2] Guangzhou Med Univ, KingMed Sch Lab Med, Guangzhou 510300, Guangdong, Peoples R China
[3] Sun Yat Sen Univ, Affiliated Hosp 1, Guangzhou 510080, Guangdong, Peoples R China
来源：
ARCHIVES OF MICROBIOLOGY | 2016年 / 198卷 / 06期
基金：
中国国家自然科学基金;
关键词：
16S rRNA; Detection; Differentiation; Legionella; Legionella pneumophila; Single-nucleotide polymorphisms; IDENTIFICATION; WATER; PCR;
D O I：
10.1007/s00203-016-1228-2
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
A PCR-based method targeting single-nucleotide polymorphisms (SNPs) in the 16S rRNA gene was developed for differential identification of Legionella pneumophila and non-Legionella pneumophila. Based on the bioinformatics analysis for 176 Legionella 16S rRNA gene fragments of 56 different Legionella species, a set of SNPs, A(628)C(629) was found to be highly specific to L. pneumophila strains. A multiplex assay was designed that was able to distinguish sites with limited sequence heterogeneity between L. pneumophila and non-L. pneumophila in the targeted 16S rRNA gene. The assay amplified a 261-bp amplicon for Legionella spp. and a set of 203- and 97-bp amplicons only specific to L. pneumophila species. Among 49 ATCC strains and 284 Legionella isolates from environmental water and clinical samples, 100 % of L. pneumophila and non-L. pneumophila strains were correctly identified and differentiated by this assay. The assay presents a more rapid, sensitive and alternative method to the currently available PCR-sequencing detection and differentiation method.
引用
收藏
页码：591 / 594
页数：4
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