Jauk V, Neubauer C, Szölgyényi W, Vasicek L
Clinic for Poultry and Pet Birds, Veterinary University Vienna, Austria.
Avian Pathol. 2003 Feb;32(1):33-7. doi: 10.1080/0307945021000070697.
An identification scheme based on restriction fragment length polymorphism of polymerase chain reaction products (PCR-RFLP) was developed to differentiate isolates of the genera Campylobacter, Arcobacter and Helicobacter. Based on the 16S rRNA gene of these genera, PCR amplified a 1216-bp fragment. The amplicons were digested with the restriction enzymes RsaI and EcoRV. Additional differentiation was obtained using a PCR-assay based on the hippuricase gene. Genotyping was performed on several reference strains from the National Collection of Typing Culture (NCTC), London, and on 130 field isolates. In parallel, a phenotypic differentiation was performed, in order to compare the results. In 119 cases (91.5%) the results obtained from the genotypic characterization were concordant with those from phenotypic testing. Co-infections with Campylobacter jejuni and Campylobacter coli in two samples and seven hippurate-negative C. jejuni-strains were identified by the genotypic method. Furthermore, PCR-RFLP assays identified an atypical isolate as Campylobacter fetus/hyointestinalis.
开发了一种基于聚合酶链反应产物限制性片段长度多态性(PCR-RFLP)的鉴定方法,以区分弯曲杆菌属、弓形杆菌属和幽门螺杆菌属的分离株。基于这些属的16S rRNA基因,PCR扩增出一个1216bp的片段。扩增产物用限制性内切酶RsaI和EcoRV进行消化。使用基于马尿酸酶基因的PCR检测获得了进一步的区分。对来自伦敦国家典型培养物保藏中心(NCTC)的几种参考菌株和130株现场分离株进行了基因分型。同时,进行了表型区分,以便比较结果。在119例(91.5%)中,基因分型鉴定结果与表型检测结果一致。通过基因分型方法鉴定出两个样本中存在空肠弯曲菌和结肠弯曲菌的混合感染,以及七株马尿酸盐阴性空肠弯曲菌菌株。此外,PCR-RFLP检测将一株非典型分离株鉴定为胎儿弯曲菌/猪肠弯曲菌。