Sacchi C T, de Lemos A P, Casagrande S T, Mori A M, de Almeida C L
Bacteriology Division, Adolfo Lutz Institute, São Paulo, Brazil.
Rev Inst Med Trop Sao Paulo. 1995 Jul-Aug;37(4):291-6. doi: 10.1590/s0036-46651995000400002.
In the present study we report the results of an analysis, based on ribotyping of Corynebacterium diphtheriae intermedius strains isolated from a 9 years old child with clinical diphtheria and his 5 contacts. Quantitative analysis of RFLPs of rRNA was used to determine relatedness of these 7 C.diphtheriae strains providing support data in the diphtheria epidemiology. We have also tested those strains for toxigenicity in vitro by using the Elek's gel diffusion method and in vivo by using cell culture method on cultured monkey kidney cell (VERO cells). The hybridization results revealed that the 5 C.diphtheriae strains isolated from contacts and one isolated from the clinical case (nose case strain) had identical RFLP patterns with all 4 restriction endonucleases used, ribotype B. The genetic distance from this ribotype and ribotype A (throat case strain), that we initially assumed to be responsible for the illness of the patient, was of 0.450 showing poor genetic correlation among these two ribotypes. We found no significant differences concerned to the toxin production by using the cell culture method. In conclusion, the use of RFLPs of rRNA gene was successful in detecting minor differences in closely related toxigenic C.diphtheriae intermedius strains and providing information about genetic relationships among them.
在本研究中,我们报告了一项分析结果,该分析基于从一名患有临床白喉的9岁儿童及其5名接触者中分离出的中间型白喉棒状杆菌菌株的核糖分型。对rRNA的限制性片段长度多态性(RFLP)进行定量分析,以确定这7株白喉棒状杆菌菌株的亲缘关系,为白喉流行病学提供支持数据。我们还通过Elek凝胶扩散法在体外以及在培养的猴肾细胞(VERO细胞)上使用细胞培养法在体内测试了这些菌株的产毒性。杂交结果显示,从接触者中分离出的5株白喉棒状杆菌菌株以及从临床病例(鼻部病例菌株)中分离出的1株菌株,与所使用的所有4种限制性内切酶的RFLP模式相同,核糖型为B。我们最初认为导致患者患病的这种核糖型与核糖型A(喉部病例菌株)之间的遗传距离为0.450,表明这两种核糖型之间的遗传相关性较差。通过细胞培养法,我们发现毒素产生方面没有显著差异。总之,使用rRNA基因的RFLP成功检测出密切相关的产毒性中间型白喉棒状杆菌菌株之间的微小差异,并提供了它们之间遗传关系的信息。