title: Evaluation of high-resolution melting and other molecular methods in discrimination of Lactobacillus isolates.
abstract: OBJECTIVE
  Fermented and probiotic products are mainly based on the activity of diverse Lactobacillus 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.
  
  
  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 Lactobacillus, a Lactococcus and a Streptococcus 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.
  
  
  CONCLUSIONS
  The newly developed HRM primers offer a quick and efficient tool for discrimination of lactobacilli, including very closely related Lactobacillus subspecies.
authors.affiliation: Egerfood Regional Knowledge Centre, Eszterházy Károly College, Eger, Hungary.
authors.forename: O P
authors.initials: OP
authors.lastname: Szén
authors.forename: A
authors.initials: A
authors.lastname: Kiss
authors.forename: Z
authors.initials: Z
authors.lastname: Naár
authors.forename: K
authors.initials: K
authors.lastname: Pál
mesh_terms: D017931:DNA Primers / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005516:Food Microbiology; D020459:Genes, rRNA; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S
keywords: 16S rDNA; HRM; Lactobacillus discrimination; PCR-RFLP; species-specific PCR