Premium
Evaluation of high‐resolution melting and other molecular methods in discrimination of L actobacillus isolates
Author(s) -
Szén O.P.,
Kiss A.,
Naár Z.,
Pál K.
Publication year - 2014
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12599
Subject(s) - kiss (tnc) , library science , computer science , information retrieval , computer network
Aims Fermented and probiotic products are mainly based on the activity of diverse L actobacillus species. Distinguishing of these species, especially the closely related ones might be problematic. Our aim was to compare and evaluate molecular methods that have the potential of discrimination and might serve as alternatives of traditional microbiological techniques. Methods and Results In our experiments, PCR methods using general and species‐specific bacterial primers, RFLP, sequencing and HRM assays were tested and their efficiency compared. A new universal primer pair was designed for amplification of short fragments of the 16S rDNA of six L actobacillus, a L actococcus and a S treptococcus species; furthermore, successful HRM analysis was performed on them that resulted in the separation of each species, including the almost indistinguishable Lact. paracasei ssp. paracasei and Lact. paracasei ssp. tolerans subspecies. Conclusions The results showed that HRM might be a useful, time‐ and cost‐saving one‐step tool for preliminary classification of isolates, although the use of additional techniques, like species‐specific PCR , analysis of RFLP patterns and sequencing, might be necessary for confirmation of the results. Significance and Impact of the Study The newly developed HRM primers offer a quick and efficient tool for discrimination of lactobacilli, including very closely related Lactobacillus subspecies.