Publication detail
Optimalization of HRM-PCR method for identification lactic acid bacteria from complex samples
KONEČNÁ, J. TRACHTOVÁ, Š. ŠPANOVÁ, A. RITTICH, B.
Original Title
Optimalization of HRM-PCR method for identification lactic acid bacteria from complex samples
Type
conference proceedings
Language
English
Original Abstract
Molecular based differentiation of various bacterial species is important mainly in diagnostic, epidemiological and food quality control studies, particulary where phenotype makes the identification of bacteria difficult. Molecular methods based on the amplification of 16S ribosomal RNA gene analysis are more rapid, reliable, and reproducible and have greater sensitivity. High resolution melting (HRM) curve analysis is a simple, low-ccost, closed tube method for amplicon discrimination and easy integration with real.time polymerase chain reaction (HRM-PCR). In this contribution we report rapid species identification of lactic acid bacteria using HRM-PCR in complex food samples. Three different DNA isolation methods were used in this work: phenol extraction, separation using magnetic particles and commercial kit. Three sets of primers targeted hypervariable region (V1-F - V1-R, V3-F - V3-R, V6-F - V6-R) were tested for amplification of the 16S rRNA gene. Presented method was optimized on 6 Reference collection of 6 strains was used for identification of lactic acid bacteria in food supplements Lactomax and Lactobacilly forte. Real-time PCR and HRM profiles generated reproducible results which allow species recognition in real samples. However, further research with larger strains collectionist is required to validate the sensitivity and robustness of this method.
Keywords
High resolution melt analysis, lactic acid bacteria, extraction methods
Authors
KONEČNÁ, J.; TRACHTOVÁ, Š.; ŠPANOVÁ, A.; RITTICH, B.
Released
28. 8. 2016
Location
Brno, Czech republic
ISBN
978-80-7305-777-0
Book
Book of Abstracts
Pages from
111
Pages to
111
Pages count
1
BibTex
@proceedings{BUT138054,
editor="Jana {Konečná} and Štěpánka {Trachtová} and Alena {Španová} and Bohuslav {Rittich}",
title="Optimalization of HRM-PCR method for identification lactic acid bacteria from complex samples",
year="2016",
pages="111--111",
address="Brno, Czech republic",
isbn="978-80-7305-777-0"
}