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. 1999 Sep;37(9):2772-6.
doi: 10.1128/JCM.37.9.2772-2776.1999.

PCR-Based methods for genotyping viridans group streptococci

S Alam  1 , S R Brailsford, R A Whiley, D Beighton
Affiliations

PCR-Based methods for genotyping viridans group streptococci

S Alam et al. J Clin Microbiol. 1999 Sep.

Abstract

Standard repetitive extragenic palindromic (REP)-PCR, enterobacterial repetitive intergenic consensus-PCR, and Salmonella enteritidis repetitive element-PCR methods for bacterial strain typing were performed with DNA extracted by boiling members of each of the currently recognized species of human viridans group streptococci. Each of the methods was reproducible. The unique isolates (n = 72) from 15 species of viridans group streptococci were readily distinguishable, with no two isolates showing greater than 90% per cent similarity. The majority of strains exhibited much less than 90% similarity. Isolates identical by REP-PCR were also identical by the other two methods. These PCR-based typing methods, although they do not permit determination of the species of the isolates, are simple to perform and are suitable for clinical and ecological investigations of viridans group streptococci.

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Figures

FIG. 1
FIG. 1
Representative REP-PCR patterns of viridans group streptococci. Lanes 2 to 6, S. infantis 0103, 092, 0101, JCM 10157T, and 0134, respectively; lanes 8 to 10, S. peroris 105, 091, and JCM 10158T, respectively; lanes 1, 7, and 11, molecular size markers.
FIG. 2
FIG. 2
Dendrogam showing relatedness between REP-PCR band patterns of viridans group streptococci. Bands were analyzed by applying the Dice coefficient, and the matrix was clustered by the UPGMA method.
FIG. 3
FIG. 3
Representative ERIC-PCR patterns of viridans group streptococci. Lanes 2 to 7, S. anginosus NCTC 10713T, NMH 10, PC4890, NCTC 11062, KR 687, and NCTC 8037, respectively; lanes 9 to 14, S. salivarius A385, NCTC 8606, H50, KPS1, T267, and NCTC 8618T, respectively; lanes 1, 8, and 15, molecular size markers.
FIG. 4
FIG. 4
Representative SERE-PCR patterns of viridans group streptococci. Lanes 2 to 6, S. sobrinus ATCC 33478T, OMZ 65, 279, TH62, and B13, respectively; lanes 8 to 12, S. mutans SE11, 161, KPSK2, B48, and NCTC 10449T, respectively; lanes 1, 7, and 13, molecular size markers.
FIG. 5
FIG. 5
Comparison of patterns produced by REP-PCR (a), ERIC-PCR (b), and (c) SERE-PCR with duplicate strains of S. oralis isolated from five different individual strains. Isolates from the same individual are in adjacent lanes (from left to right), and in each gel the same isolate occupies the same lane. Lanes 1, 8, and 13, molecular size markers.
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