Restriction fragment length polymorphism analysis of the flaA gene of Campylobacter jejuni for subtyping human, animal and poultry isolates.
Restriction fragment length polymorphism analysis of the flaA gene of Campylobacter jejuni for subtyping human, animal and poultry isolates.
复制标题
空肠弯曲杆菌 flaA 基因的限制性片段长度多态性分析，用于对人类、动物和家禽分离株进行亚型分析。
作者：
R. Owen;S. Leeton
采用EcoRI和PstI （EP flaA-profiling）双酶切法对233株空肠弯曲杆菌的鞭毛蛋白（flaA）基因进行PCR-RFLP分型。这些菌株代表了多种常见的Penner热稳定（HS）血清型，包括来自人、牛、羊、鸡和犬的分离株。FlaA扩增子直接从大多数菌株的细胞裂解物DNA中获得。RFLP分析显示等位基因变异明显，共鉴定出9种EP flaa型，其中最常见的是2型（32%）、3型（20%）、4型（12%）和6型（12%）。其他flaA-profiles分别代表不到10%的菌株。每种血清型的空肠C.菌株一般有一种或两种特异性相关的flaa型，尽管有些是几种血清型的特征。在不同宿主中发现具有相同flaa型的菌株。EP flaA-profiles重现性好、记录清晰、简单，实验方案快速、低成本、高通量。EP flaA-profiling方案为HS血清分型提供了一种优良的分子分型方法，并推荐参考菌株用于今后的流行病学调查。
233 strains of Campylobacter jejuni were subtyped by PCR-RFLP analysis of the flagellin (flaA) gene by double digestion with EcoRI and PstI (EP flaA-profiling). The strains represented a variety of common Penner heat stable (HS) serotypes and comprised isolates of human, bovine, ovine, chicken and canine origin. FlaA amplicons were obtained directly from DNA in cell lysates of most strains. RFLP analysis showed considerable allelic variation and nine EP flaA-types were identified of which the most common were type 2 (32%), type 3 (20%), type 4 (12%) and type 6 (12%). Other flaA-profiles each represented less than 10% of strains. C. jejuni strains of each serotype generally had one or two specifically associated flaA-types although some were features of several serotypes. Strains with the same flaA-type were found in different hosts. EP flaA-profiles were reproducible, clear and simple to record, and laboratory protocols were rapid and low cost with high throughput capacity. The EP flaA-profiling scheme provided an excellent molecular subtyping method to supplement HS serotyping, and reference strains are recommended to facilitate its use in future epidemiological investigations.