Richner S M, Meiring J, Kirby R
Department of Biochemistry and Microbiology, Rhodes University, Grahamstown, South Africa.
Electrophoresis. 1997 Aug;18(9):1570-6. doi: 10.1002/elps.1150180915.
This population genetics study was done to determine the degree of genetic diversity amongst clinical isolates of Mycobacterium tuberculosis in one of the largest provinces of South Africa. Three hundred and fifty-nine individual cultures were obtained from single patients over a nine-month period. Bacterial DNA was extracted and amplified with two arbitrary ten-mer primers, using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) technique. RAPD markers were separated by agarose gel electrophoresis, visualised with ethidium bromide, and then analysed for similarity using the GelCompar programme. The isolates were seen to fall into two major population groups. A high degree of genetic diversity was seen, with a total of 350 unique strains occurring amongst the 359 isolates. The majority of drug-resistant isolates grouped together in the smaller population group. The genetic data were also correlated with geographical locality. It was found that a slight preponderance of isolates that grouped together closely, vis. that were either genetically identical or very similar (similarity index of approximately 97%), were from areas that were considered to be geographically distant. Antibiotic resistance patterns were also examined, revealing a total of 16 different resistance profiles among the 68 drug-resistant isolates. RAPD-PCR has proved to be a cost- and time-effective technique, suitable for such a large-scale population genetics study. It has provided useful information on the genetic diversity of an organism which is responsible for an increasing amount of morbidity and mortality in South Africa's second-largest province.
这项群体遗传学研究旨在确定南非最大省份之一结核分枝杆菌临床分离株之间的遗传多样性程度。在九个月的时间里,从单个患者身上获取了359个单独的培养物。提取细菌DNA,使用随机扩增多态性DNA - 聚合酶链反应(RAPD - PCR)技术,用两种任意的十聚体引物进行扩增。RAPD标记通过琼脂糖凝胶电泳分离,用溴化乙锭染色观察,然后使用GelCompar程序分析相似性。分离株被分为两个主要群体。观察到高度的遗传多样性,在359个分离株中共有350个独特菌株。大多数耐药分离株聚集在较小的群体中。遗传数据也与地理位置相关。研究发现,紧密聚集在一起的分离株(即基因相同或非常相似,相似性指数约为97%)略微倾向于来自被认为地理位置遥远的地区。还检查了抗生素耐药模式,在68个耐药分离株中发现了总共16种不同的耐药谱。RAPD - PCR已被证明是一种经济高效的技术,适用于如此大规模的群体遗传学研究。它为一种在南非第二大省导致发病率和死亡率不断上升的生物体的遗传多样性提供了有用信息。