Demkin V V, Rydkina E B, Likhoded L Y, Ignatovich V F, Genig V A, Balayeva N M
Gamaleya Research Institute of Epidemiology and Microbiology, Moscow, Russia.
Acta Virol. 1994 Apr;38(2):65-70.
Southern blot analysis of HindIII-cleaved rickettsial DNA was used for genotypic characterization of the typhus group (TG) species (R. prowazekii, R. typhi, R. canada) and a few species of the spotted fever group (SFG) rickettsiae (R. sibirica, R. conorii, R. akari). Four different DNA probes were employed. PBH11 and PBH13 probes were morphospecific HindIII fragments of R. prowazekii DNA. MW218 probe contained the gene for 51 K antigen and MW264 probe contained the citrate synthase gene of R. prowazekii. All the probes hybridized with the tested TG and SFG rickettsial DNAs, forming from 1 to 5 bands, but they did not with R. tsutsugamushi or C. burnetii DNAs. All the probes demonstrated specific hybridization patterns with TG species and R. akari. PBH11, PBH13 and MW264 probes clearly distinguished R. sibirica and R. conorii from the other tested rickettsiae, but not from each other. However, these two species differed slightly with MW218 probe. Several strains of each species were analyzed in this way and except for strains of R. conorii identical intraspecies patterns were obtained. These data lead us to consider the obtained hybridization patterns as criteria for genotypic identification.
采用HindIII酶切立克次氏体DNA的Southern印迹分析,对斑疹伤寒群(TG)物种(普氏立克次氏体、鼠型斑疹伤寒立克次氏体、加拿大立克次氏体)和斑点热群(SFG)立克次氏体的几个物种(西伯利亚立克次氏体、康氏立克次氏体、小蛛立克次氏体)进行基因分型鉴定。使用了四种不同的DNA探针。PBH11和PBH13探针是普氏立克次氏体DNA的形态特异性HindIII片段。MW218探针包含51K抗原基因,MW264探针包含普氏立克次氏体的柠檬酸合酶基因。所有探针均与受试的TG和SFG立克次氏体DNA杂交,形成1至5条带,但与恙虫病东方体或贝氏柯克斯体DNA不杂交。所有探针均显示出与TG物种和小蛛立克次氏体的特异性杂交模式。PBH11、PBH13和MW264探针能清楚地将西伯利亚立克次氏体和康氏立克次氏体与其他受试立克次氏体区分开来,但无法区分这两者。然而,这两个物种在MW218探针检测下略有差异。以这种方式分析了每个物种的几个菌株,除了康氏立克次氏体菌株外,均获得了种内一致的模式。这些数据使我们将获得的杂交模式视为基因分型鉴定的标准。