Use of tuf Sequences for Genus-Specific PCR Detection and Phylogenetic Analysis of 28 Streptococcal Species

摘要: A 761-bp portion of the tuf gene (encoding elongation factor Tu) from 28 clinically relevant streptococcal species was obtained by sequencing amplicons generated using broad-range PCR primers. These sequences were used to select Streptococcus-specific primers and perform phylogenetic analysis. The specificity assay verified 102 different bacterial species, including species. Genomic DNA purified all efficiently detected, whereas there no amplification with 72 74 nonstreptococcal tested. There cross-amplification DNAs Enterococcus durans Lactococcus lactis. However, 15 31% nucleotide sequence divergence in these two compared any provides ample allow development internal probes specific streptococci. highly sensitive for tested (i.e., detection limit 1 10 genome copies per PCR). data also extensive analysis, which generally agreement phylogeny determined on basis 16S rRNA data. provided a better discrimination at level that should be particularly useful identification very closely related In conclusion, appears more suitable than ribosomal RNA diagnostic assays because its higher species-specific genetic divergence.