M protein gene typing of Streptococcus pyogenes by nonradioactively labeled oligonucleotide probes.
A new approach for the typing of Streptococcus pyogenes is described. Oligonucleotide probes of 30 nucleotides in length were derived from currently known sequences of the N-terminal regions of M protein genes (emm genes). The oligonucleotides were labeled with digoxigenin-dUTP and hybridized to dot-blotted genomic DNA from 116 group A streptococcal strains of serotypes M-1, M-2, M-3, M-5, M-6, M-12, M-18, M-19, M-24, and M-49. Hybridization reactions were visualized with a chemiluminescent substrate. In comparison with conventional serological typing of expressed M proteins, the binding of the probes to the corresponding emm genes exhibited 100% sensitivity and specificity. The results emphasize the high degree of type-specific conservation of the N-terminal regions of emm genes from reference strains and epidemiologically unrelated U.S. and European clinical isolates. The existence of two distinct genetic subgroups among eight investigated M-49 strains was unequivocally shown by hybridization assays and further confirmed by nucleotide sequence data obtained from four selected M-49 strains. Because oligonucleotide probes are relatively easy to prepare, easy to handle, and known to give consistent interlaboratory results, the "oligotyping" technique appears to offer potential advantages over conventional serological typing methods.
Kaufhold
A
A
Institute of Medical Microbiology, Technical University (RWTH) Aachen, Germany.
Podbielski
A
A
Johnson
D R
DR
Kaplan
E L
EL
Lütticken
R
R
D000942:Antigens, Bacterial / Q000145:classification / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D015139:Blotting, Southern; D002352:Carrier Proteins; D005798:Genes, Bacterial / Q000235:genetics*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D012015:Reference Standards; D012680:Sensitivity and Specificity; D017385:Sequence Homology; D013297:Streptococcus pyogenes / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; C014689:streptococcal M protein