Molecular characterization of invasive and noninvasive Campylobacter jejuni and Campylobacter coli isolates
Molecular characterization of invasive and noninvasive Campylobacter jejuni and Campylobacter coli isolates
复制标题
DOI：
10.1128/jcm.39.4.1353-1359.2001
复制
发表时间：
2001-04-01
影响因子：
9.4
通讯作者：
Pickering, LK
中科院分区：
文献类型：
--
作者：
Carvalho, ACT;Ruiz-Palacios, GM;Pickering, LK
空肠弯曲杆菌是全球细菌性腹泻的最常见原因之一，也是食源性疾病的主要细菌原因。对上皮细胞的粘附和侵袭是弯曲菌腹泻的重要致病机制。应用随机扩增多态性DNA技术（RAPD）对腹泻患儿和非腹泻患儿的侵袭性和非侵袭性弯曲菌进行了分子特征分析。一个独特的RAPD图谱与1.6 lib的DNA带之间的侵袭性（63%）比非侵袭性（16%）弯曲杆菌菌株（P = 0.000005）观察到显着更频繁。将1.6kb的条带命名为入侵相关标记（IAM）。使用专门设计的引物，518 bp的IAM基因座的片段扩增85%的侵入性和20%的非侵入性菌株（P = 0.0000000）。PCR-限制性片段长度多态性分析，扩增整个IAM位点的分子分型显示HindIII限制性片段多态性模式主要与侵袭性菌株。虽然RAPD指纹图谱的聚类分析显示菌株之间的遗传多样性，两个主要的集群被确定。第一簇包括致病性和侵袭性明显更高的分离株，而第二簇则将大多数非致病性、非侵袭性分离株分组。这些数据表明，大多数的侵袭性弯曲杆菌菌株可以区分从非侵袭性菌株的RAPD分析和FCR使用特异性引物扩增的IAM位点的片段。
Campylobacter jejuni is one of the most common causes of bacterial diarrhea worldwide and is the primary bacterial cause of food-borne illness. Adherence to and invasion of epithelial cells are the most important pathogenic mechanisms of Campylobacter diarrhea, Molecular characterization of invasive and noninvasive Campylobacter isolates from children with diarrhea and symptom-free children was performed by random amplified polymorphic DNA techniques (RAPD). A distinct RAPD profile with a DNA band of 1.6 lib was observed significantly more frequently among invasive (63%) than among noninvasive (16%) Campylobacter isolates (P = 0.000005). The 1.6-kb band was named the invasion-associated marker (IAM). Using specifically designed primers, a fragment of 518 bp of the iam locus was amplified in 85% of invasive and 20% of noninvasive strains (P = 0.0000000). Molecular typing with a PCR-restriction fragment length polymorphism assay which amplified the entire iam locus showed a HindIII restriction fragment polymorphism pattern associated mainly with invasive strains. Although cluster analysis of the RAPD fingerprinting showed genetic diversity among strains, two main clusters were identified. Cluster I comprised significantly more pathogenic and invasive isolates, while cluster II grouped the majority of nonpathogenic, noninvasive isolates. These data indicate that most of the invasive Campylobacter strains could be differentiated from noninvasive isolates by RAPD analysis and FCR using specific primers that amplify a fragment of the iam locus.