Kaufhold A, Podbielski A, Baumgarten G, Blokpoel M, Top J, Schouls L
Institute of Medical Microbiology, Technical University (RWTH) Aachen, FRG.
FEMS Microbiol Lett. 1994 Jun 1;119(1-2):19-25. doi: 10.1111/j.1574-6968.1994.tb06861.x.
Because of the allelic variations within the M protein gene (emm gene) of group A streptococci, reliable typing of this important human pathogen can be accomplished by the use of emm gene-specific oligonucleotide probes. Two technical modifications (a reverse dot blot and a reverse line blot hybridization assay) of a novel approach for the type-specific identification of emm genes have been developed. Both procedures involved amplification of an emm gene by polymerase chain reaction. The non-radioactively labeled amplicon was subsequently hybridized to a membrane carrying an array of immobilized emm gene-specific oligonucleotide probes, thus allowing the simultaneous analysis of the gene polymorphism in a single hybridization reaction. The feasibility of these rapid and easy to perform methods was shown for the unequivocal identification of reference strains and clinical isolates belonging to 16 different M serotypes.
由于A组链球菌M蛋白基因（emm基因）内存在等位基因变异，使用emm基因特异性寡核苷酸探针可实现对这种重要人类病原体的可靠分型。已开发出一种用于emm基因型特异性鉴定的新方法的两种技术改进方法（反向斑点印迹法和反向线印迹杂交分析）。这两种方法都涉及通过聚合酶链反应扩增emm基因。随后将非放射性标记的扩增子与携带一系列固定化emm基因特异性寡核苷酸探针的膜杂交，从而能够在单个杂交反应中同时分析基因多态性。这些快速且易于操作的方法对于明确鉴定属于16种不同M血清型的参考菌株和临床分离株的可行性已得到证明。
