Evaluation of (GTG)₅-PCR for identification ofEnterococcusspp.

Abstract

A set of reference strains and a group of previously unidentified enterococci were analysed by rep-PCR with the (GTG)(5) primer to evaluate the discriminatory power and suitability of this method for typing and identification of enterococcal species. A total of 49 strains representing all validly described species were obtained from bacterial collections. For more extensive evaluation of this identification approach 112 well-defined and identified enterococci isolated from bryndza cheese were tested. The (GTG)(5)-PCR fingerprinting assigned all strains into well-differentiated clusters representing individual species. Subsequently, a group including 44 unidentified enterococci isolated from surface waters was analysed to evaluate this method for identification of unknown isolates. Obtained band patterns allowed us to identify all the strains clearly to the species level. This study proved that rep-PCR with (GTG)(5) primer is a reliable and fast method for species identification of enterococci.

Genotypic characterization and safety assessment of lactic acid bacteria from indigenous African fermented food products

“…The reproducibility and discriminatory power of rep-PCR (GTG) 5 in typing at species and subspecies level have previously been reported [8,43-45] and also in the present study the technique proved useful for genotypic fingerprinting and grouping.…”

Assessment of Coagulase‐Negative Staphylococci and Lactic Acid Bacteria Isolated from Portuguese Dry Fermented Sausages as Potential Starters Based on Their Biogenic Amine Profile

“…were isolated on Manitol Salt Agar (MSA, Scharlau, Spain) at 37°C for 48 hr and species were identified by PCR according to Morot-Bizot, Talon, and Leroy-Setrin (2003) and Blaiotta, Casaburi, and Villani (2005). Genomic typing was performed by PCR-fingerprinting using the primers (GTG) 5, M13 and OPC-19 in independent reactions, according to Svec et al (2005) and Ribeiro et al (2011). A collection of selected strains was obtained according to their technological or protective characteristics (bacteriocinogenic activity) but with different genomic profiles (n = 9 staphylococci, n = 22 lactobacilli, n = 6 enterococci) being tested regarding the decarboxylase activity and the ability to produce biogenic amines.…”

Presence of Different Bacterial Species in Thermal Sources and Novelty in Their Industrial Enzyme Productions

“…Test isolates were subjected to rep-PCR with the special primers of (GTG) 5 and BOX elements to obtain genomic fingerprinting 4,6,20 . To obtain the PCR products, 50 The 16S rRNA of the isolates was amplified by Polymerase Chain Reaction (PCR) with the forward primer 27-F:(5' AGAGTTTGATYMTGGCTCAG3') and the reverse primer 1492-R:(5' GGTTACCTTGTTACGACTT 3') 5 . The amplified fragments were cloned into E. coli JM101 strain with a vector system (pGEM-T, Promega, the UK) and the clones were sequenced (Macrogen, Amsterdam, the Netherlands).…”