Identification of group A type 1 streptococcal M protein gene by a non-radioactive oligonucleotide detection method.
An oligonucleotide probe of 30 nucleotides length has been constructed, spanning the codons of amino acids 2 to 11 of the mature M1 protein of group A streptococci (Streptococcus pyogenes). It was labeled with digoxigenin-dUTP and visualized after hybridization with an anti-digoxigenin-antibody/alkaline phosphatase conjugate. It definitely detected the emm1 gene in dot-blotted 20-micrograms amounts of total nucleoid acid extracts. When tested with 27 type M1 group A streptococci of several epidemiologically unrelated outbreaks and 24 non-M1 strains, its sensitivity and specificity of detection reached 100% even at a hybridization temperature 35 degrees C below the calculated Tm. A detailed protocol for the construction and use of this oligonucleotide probe is given.
Podbielski
A
A
Institute of Medical Microbiology, Klinikum der RWTH, Aachen, Federal Republic of Germany.
Kühnemund
O
O
Lütticken
R
R
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005798:Genes, Bacterial; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D013297:Streptococcus pyogenes / Q000235:genetics*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D015342:DNA Probes; D004269:DNA, Bacterial; C014689:streptococcal M protein