Targeting single-nucleotide polymorphisms in the 16S rRNA gene to detect and differentiate Legionella pneumophila and non-Legionella pneumophila species.
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.
Guangzhou KingMed Center for Clinical Laboratory, No.10, Luoxuan 3 Road, Guangzhou International Bio-island, Guangzhou, 510300, China.|KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 510300, China.|The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China.
Xiao-Yong
XY
Zhan
Guangzhou KingMed Center for Clinical Laboratory, No.10, Luoxuan 3 Road, Guangzhou International Bio-island, Guangzhou, 510300, China.|KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 510300, China.
Chao-Hui
CH
Hu
Guangzhou KingMed Center for Clinical Laboratory, No.10, Luoxuan 3 Road, Guangzhou International Bio-island, Guangzhou, 510300, China. zqy@kingmed.com.cn.|KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 510300, China. zqy@kingmed.com.cn.
Qing-Yi
QY
Zhu
D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D016952:Legionella pneumophila / Q000145:classification* / Q000235:genetics*; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S
16S rRNA; Detection; Differentiation; Legionella; Legionella pneumophila; Single-nucleotide polymorphisms