Kormilitsyna M I, Meshcheriakova I S, Mikhaĭlova T V
Mol Gen Mikrobiol Virusol. 2013(3):22-5.
Typing of Francisella strains collection by means of PCR on the basis of tul4 and RDI genes was carried out. The identification of the species and subspecies of the 112 strains of Francisella tularensis was reashed. The PCR on DNA-targets loci of type IV pili genes: pilA, pilE2, pilE3, pilE4, pilE5, pilF, pilT, pilD and pilQ for differentiation of F. tularensis strains on virulence was carried out. It was demonstrated the possibility of differentiation of F. tularensis strains in PCR (primers A-B) on the basis of the revelation of the gene pilA in virulent strains of third subspecies F. tularensis and F. tularensis subsp. novicida. This gene pilA was not detected in the vaccine strain 15/10 and its variants, as well as in the most of avirulent F. tularensis subsp. holarctica strains. However, the fragment gene pilA was found in the attenuated strains F. tularensis subsp. tularensis and mediasiatica. It was not revealed any differences on other targets of pili genes between of F. tularensis strains, with the exception of the strain F. tularensis subsp. novicida Utah 112, which had not a fragment of the gene pilE2. The use of PCR to target the locus of the pilA gene allows to discriminate virulent F. tularensis subsp. holarctica strains from the vaccine 15/10, its variants and avirulent strains.
通过基于tul4和RDI基因的PCR对弗朗西斯菌菌株进行分型。完成了对112株土拉弗朗西斯菌的种和亚种鉴定。针对IV型菌毛基因的DNA靶点位点(pilA、pilE2、pilE3、pilE4、pilE5、pilF、pilT、pilD和pilQ)进行PCR,以区分土拉弗朗西斯菌菌株的毒力。结果表明,基于在土拉弗朗西斯菌第三亚种和土拉弗朗西斯菌新凶手亚种的强毒株中发现pilA基因,可在PCR(引物A - B)中区分土拉弗朗西斯菌菌株。在疫苗株15/10及其变体以及大多数无毒力的土拉弗朗西斯菌全北区亚种菌株中未检测到该pilA基因。然而,在减毒株土拉弗朗西斯菌土拉亚种和中亚细亚亚种中发现了pilA基因片段。除土拉弗朗西斯菌新凶手亚种犹他112株外,土拉弗朗西斯菌菌株在菌毛基因的其他靶点上未发现差异,该株没有pilE2基因片段。使用PCR靶向pilA基因位点可区分有毒力的土拉弗朗西斯菌全北区亚种菌株与疫苗株15/10、其变体和无毒力菌株。