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使用(GTG)5寡核苷酸作为雷氏弯曲菌分型的随机扩增多态性DNA引物。

The use of (GTG)5 oligonucleotide as an RAPD primer to type Campylobacter concisus.

作者信息

Matsheka M I, Lastovica A J, Zappe H, Elisha B G

机构信息

Department of Medical Microbiology, University of Cape Town, Cape Town, South Africa.

出版信息

Lett Appl Microbiol. 2006 Jun;42(6):600-5. doi: 10.1111/j.1472-765X.2006.01900.x.

Abstract

AIM

DNA fingerprinting using (GTG)(5) oligonucleotide as a primer in a random amplified polymorphic DNA (RAPD) assay was assessed by typing isolates of Campylobacter concisus strains, collected over a period of 8 years.

METHODS AND RESULTS

RAPD analysis using the (GTG)(5) oligonucleotide as a primer was used to type 100 isolates of C. concisus comprising mostly isolates from children with diarrhoea. Using this method, 86% of the isolates were found to be genotypically diverse. Of these heterogeneous isolates, 25 of the strains were also shown to be genetically distinct in a previous study using pulsed field gel electrophoresis. The remaining isolates (14) could be classified into five profile groups based on the DNA fingerprinting patterns. The assay successfully identified epidemiologically linked strains from the unrelated genetically diverse pool of strains.

CONCLUSIONS

Laboratory RADP typing using the (GTG)(5) primer proved to be useful in distinguishing related strains of C. concisus from a large pool of unrelated strains of this organism.

SIGNIFICANCE AND IMPACT OF THE STUDY

RAPD typing using (GTG)(5) is a simple method that could be used to investigate the epidemiology of C. concisus. The results suggest that homologous lineages of C. concisus may exist within an otherwise heterogeneous species complex. However, these data need to be confirmed using a more robust typing method.

摘要

目的

通过对8年期间收集的简明弯曲杆菌菌株分离株进行分型,评估以(GTG)₅寡核苷酸为引物的随机扩增多态性DNA(RAPD)分析用于DNA指纹图谱分析的情况。

方法与结果

使用以(GTG)₅寡核苷酸为引物的RAPD分析对100株简明弯曲杆菌分离株进行分型,这些分离株大多来自腹泻儿童。使用该方法,发现86%的分离株基因型不同。在这些异质分离株中,25株在先前使用脉冲场凝胶电泳的研究中也显示出基因差异。其余分离株(14株)可根据DNA指纹图谱分为五个谱型组。该分析成功地从无关的基因多样的菌株库中鉴定出了具有流行病学关联的菌株。

结论

使用(GTG)₅引物进行实验室RAPD分型被证明有助于从大量无关的该生物体菌株中区分出相关的简明弯曲杆菌菌株。

研究的意义和影响

使用(GTG)₅进行RAPD分型是一种可用于调查简明弯曲杆菌流行病学的简单方法。结果表明,在其他方面为异质的物种复合体中可能存在简明弯曲杆菌的同源谱系。然而,这些数据需要使用更可靠的分型方法进行确认。

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