Cavé H, Bingen E, Elion J, Denamur E
Laboratoire de Biochimie Génétique, Hôpital Robert Debré, Paris.
Res Microbiol. 1994 Feb;145(2):141-50. doi: 10.1016/0923-2508(94)90007-8.
Fifty-nine Escherichia coli strains isolated from 54 unrelated patients over a six-year period, as well as the reference strain of the species, were studied by analysis of randomly amplified polymorphic DNA (RAPD) to assess the usefulness of this genotyping approach in molecular epidemiology. Using a 10-mer oligonucleotide primer, 28 different RAPD fingerprints were distinguished among the 60 strains previously delineated in 36 ribotypes by EcoRI and HindIII digests. The patterns were reproducible and stable after in vitro and in vivo studies. Some strains harbouring an identical ribotype exhibited distinct RAPD fingerprints. Thus, these data illustrate the usefulness of the association of two genetic markers in assessing the relationship between strains. Interestingly, among the RAPD patterns, several bands were present only in the highly virulent carboxylesterase type B2 strains, which are more homogeneous than the carboxylesterase type B1 strains. Because of its simplicity and rapidity, RAPD analysis appears to be a highly valuable tool for studying E. coli molecular epidemiology.
对在六年时间里从54名无亲缘关系的患者中分离出的59株大肠杆菌菌株以及该菌种的参考菌株,通过随机扩增多态性DNA(RAPD)分析进行研究,以评估这种基因分型方法在分子流行病学中的实用性。使用一种10聚体寡核苷酸引物,在先前通过EcoRI和HindIII酶切鉴定为36种核糖型的60株菌株中区分出了28种不同的RAPD指纹图谱。这些图谱在体外和体内研究后具有可重复性和稳定性。一些具有相同核糖型的菌株表现出不同的RAPD指纹图谱。因此,这些数据说明了两种遗传标记联合使用在评估菌株间关系方面的实用性。有趣的是,在RAPD图谱中,有几条带仅出现在高毒力的B2型羧酸酯酶菌株中,这些菌株比B1型羧酸酯酶菌株更为同源。由于其简单性和快速性,RAPD分析似乎是研究大肠杆菌分子流行病学的一种极有价值的工具。