SLANÝ, Michal, Vanerkova MARTINA, Eva NĚMCOVÁ, Barbora ŽALOUDÍKOVÁ, Filip RŮŽIČKA and Tomáš FREIBERGER. Differentiation of Staphylococcus spp. by High-Resolution Melting Analysis. Canadian Journal of Microbiology. Ottawa, Canada: N R C Research Press, 2010, vol. 56, No 12, p. 1040-1049. ISSN 0008-4166. Available from: https://dx.doi.org/10.1139/W10-091.
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Basic information
Original name Differentiation of Staphylococcus spp. by High-Resolution Melting Analysis
Authors SLANÝ, Michal (203 Czech Republic, belonging to the institution), Vanerkova MARTINA (203 Czech Republic), Eva NĚMCOVÁ (203 Czech Republic, belonging to the institution), Barbora ŽALOUDÍKOVÁ (203 Czech Republic, belonging to the institution), Filip RŮŽIČKA (203 Czech Republic, belonging to the institution) and Tomáš FREIBERGER (203 Czech Republic, guarantor, belonging to the institution).
Edition Canadian Journal of Microbiology, Ottawa, Canada, N R C Research Press, 2010, 0008-4166.
Other information
Original language English
Type of outcome Article in a journal
Field of Study Genetics and molecular biology
Country of publisher Canada
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 1.235
RIV identification code RIV/00216224:14110/10:00051710
Organization unit Faculty of Medicine
Doi http://dx.doi.org/10.1139/W10-091
UT WoS 000285556000008
Keywords in English HRMA; real-time PCR; 16S rRNA; bacterial detection
Tags International impact
Changed by Changed by: Mgr. Michal Petr, učo 65024. Changed: 10/4/2012 08:32.
Abstract
High-resolution melting analysis (HRMA) is a fast high-throughput method to scan for sequence variations in a target gene. The aim of this study was to test the potential of HRMA to distinguish particular bacterial species of the Staphylococcus genus even when using a broad-range PCR within the 16S rRNA gene where sequence differences are minimal. Genomic DNA samples isolated from 12 reference strains were subjected to a real-time PCR amplification of the 16S rRNA gene in the presence of fluorescent dye, followed by HRMA. Melting profiles were used as molecular fingerprints for bacterial species differentiation. HRMA of S. saprophyticus and S. xylosus resulted in undistinguishable profiles because of their identical sequences in the analyzed 16S rRNA region. The remaining reference strains were fully differentiated either directly or via high-resolution plots obtained by heteroduplex formation between coamplified PCR products of the tested staphylococcal strain and phylogenetically unrelated strain.
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