Premium
Random amplified polymorphic DNA and amplified ribosomal DNA spacer polymorphism : powerful methods to differentiate Streptococcus thermophilus strains
Author(s) -
Giancarlo Moschetti,
Giuseppe Blaiotta,
Maria Aponte,
Pasquale Catzeddu,
Francesco Villani,
Pietrino Deiana,
Salvatore Coppola
Publication year - 1998
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 - 10.1046/j.1365-2672.1998.00461.x
Subject(s) - rapd , biology , streptococcus thermophilus , streptococcus salivarius , polymerase chain reaction , genetics , typing , primer (cosmetics) , ribotyping , pulsed field gel electrophoresis , genomic dna , dna profiling , restriction fragment length polymorphism , ribosomal dna , microbiology and biotechnology , ribosomal rna , dna , streptococcus , genotype , genetic diversity , bacteria , gene , phylogenetics , population , lactobacillus , chemistry , demography , organic chemistry , sociology
Streptococci from different collections and dairy materials were characterized by conventional and molecular methods. After amplification of the 16S–23S rDNA spacer region, all the strains referable to the genus Streptococcus exhibited a single polymerase chain reaction (PCR) product, allowing their differentiation from enterococci. Cleaving this PCR product with Hae III, two different restriction patterns could be observed, allowing Streptococcus salivarius DSM 20560 T , Strep. thermophilus NCDO 822 and two strains of Streptococcus spp. to be gathered in one group and all the other strains in another. In order to achieve strain typing, all the cultures were investigated by random amplified polymorphic DNA (RAPD)‐PCR analysis employing two selected primers. The results were treated by cluster analysis, appearing significantly consistent with both the taxonomic position and the origin of the strains. Pulsed‐field gel electrophoresis (PFGE) of Sma I digests of the genomic DNA from 11 representative strains with decreasing levels of RAPD similarity allowed their diversity to be confirmed, even though RAPD‐PCR proved to be less discriminating than PFGE analysis. The results are discussed with reference to the capability of the analytical procedures used to aid both identification and strain typing of streptococci, as well as the taxonomic structure of the species Strep. thermophilus.