Kassama Yankuba, Rooney Paul J, Goodacre Royston
Institute of Biological Sciences, University of Wales, Aberystwyth, Ceredigion, SY23 3DD, United Kingdom.
J Clin Microbiol. 2002 Aug;40(8):2795-800. doi: 10.1128/JCM.40.8.2795-2800.2002.
The ability of the fluorescent amplified fragment length polymorphism (FAFLP) technique to identify bacterial isolates from urinary tract infections (UTIs) was investigated. FAFLP was carried out using the single primer combination MseI plus CT and EcoRI plus 0, and information-rich FAFLP profiles were generated from all 69 UTI isolates studied, which comprised both gram-negative and gram-positive bacteria encompassing eight genera. The genetic relatedness of these 69 bacteria was determined by cluster analysis, and this revealed eight main groups corresponding to the eight bacterial genera. Finer discrimination on the same dendrogram showed species and subspecies differentiations, thus demonstrating the potential of FAFLP for describing a wide diversity range within microbial populations. The interpretation of FAFLP profiles is often complicated because it relies upon the investigator interpreting dendrograms; this process may be subjective if the tree is complicated, particularly if it includes polytomies (unresolved nodes). Therefore, we have developed a method based on Bayes' theorem for the identification of bacteria against an FAFLP probabilistic identification matrix. Thus, FAFLP is suitable for the objective identification of causal agents of UTI, and the procedure offers great potential in the clinical laboratory.
研究了荧光扩增片段长度多态性(FAFLP)技术识别尿路感染(UTI)细菌分离株的能力。使用单引物组合MseI加CT和EcoRI加0进行FAFLP分析,从所研究的全部69株UTI分离株中生成了信息丰富的FAFLP图谱,这些分离株包括革兰氏阴性菌和革兰氏阳性菌,涵盖八个属。通过聚类分析确定了这69种细菌的遗传相关性,结果显示有八个主要类群,分别对应八个细菌属。在同一棵树状图上进行更精细的区分显示出了种和亚种的差异,从而证明了FAFLP在描述微生物群体内广泛多样性方面的潜力。FAFLP图谱的解读通常很复杂,因为它依赖于研究人员对树状图的解读;如果树状图很复杂,尤其是包含多歧分支(未解析节点)时,这个过程可能会带有主观性。因此,我们基于贝叶斯定理开发了一种方法,用于根据FAFLP概率识别矩阵鉴定细菌。因此,FAFLP适用于客观鉴定UTI的病原体,该方法在临床实验室具有很大潜力。