Applying a PCR-based open-reading frame typing method for easy genotyping and molecular epidemiological analysis of Pseudomonas aeruginosa
| Autor: | M. Aoki, H. Baba, Eriko Hosoba, Y. Iinuma, Masakado Matsumoto, Masahiro Suzuki, Kouju Yamada |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Genomic Islands Genotype 030106 microbiology Molecular Sequence Data Biology medicine.disease_cause Applied Microbiology and Biotechnology Polymerase Chain Reaction beta-Lactamases law.invention 03 medical and health sciences Open Reading Frames Bacterial Proteins law Pulsed-field gel electrophoresis medicine Humans Pseudomonas Infections Typing ORFS Genotyping Polymerase chain reaction Genetics Cross Infection Molecular Epidemiology Molecular epidemiology Pseudomonas aeruginosa General Medicine Multilocus sequence typing Biotechnology |
| Zdroj: | Journal of applied microbiology. 120(2) |
| ISSN: | 1365-2672 |
| Popis: | AIMS Molecular epidemiological techniques, such as pulsed-field gel electrophoresis (PFGE), or multilocus sequence typing (MLST) have facilitated our understanding of the transmission routes of nosocomial infections by Pseudomonas aeruginosa. However, they are time consuming and technically demanding. To perform molecular epidemiological analysis in a standard microbiology laboratory, we aimed to develop a simpler and effective molecular epidemiological technique based on the open-reading frame (ORF) distribution patterns detected by PCR, which we call PCR-based ORF typing (POT). METHODS AND RESULTS Ten ORFs from genomic islets, five ORFs from genomic islands, and the metallo-β-lactamases (MBLs) blaIMP and blaVIM were selected by comparing the whole-genome sequences of different Ps. aeruginosa strains (PAO1, PA7, UCBPP-PA14 and LESB58). These 17 ORFs were detected, along with a Ps. aeruginosa marker, using 9-plex and 10-plex PCR systems. The genotypes in the POT were compared to those obtained by using PFGE and MLST. CONCLUSIONS Using the POT method, molecular epidemiological analyses of Ps. aeruginosa can be completed in 4 h. SIGNIFICANCE AND IMPACT OF THE STUDY Since this method is very easy to perform, even in standard clinical laboratories, it could be a valuable tool for monitoring daily infection control measures. |
| Databáze: | OpenAIRE |
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