Targeting single-nucleotide polymorphisms in the 16S rRNA gene to detect and differentiate Legionella pneumophila and non-Legionella pneumophila species
| Autor: | Qing-Yi Zhu, Xiao-Yong Zhan, Chao-Hui Hu |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
DNA Bacterial Bioinformatics analysis Legionella 030106 microbiology Single-nucleotide polymorphism Biochemistry Microbiology Legionella pneumophila Polymerase Chain Reaction Polymorphism Single Nucleotide 03 medical and health sciences RNA Ribosomal 16S Genetics Humans Multiplex Molecular Biology Gene biology General Medicine Amplicon bacterial infections and mycoses 16S ribosomal RNA biology.organism_classification respiratory tract diseases Molecular Typing 030104 developmental biology bacteria |
| Zdroj: | Archives of microbiology. 198(6) |
| ISSN: | 1432-072X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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