Matsheka M I, Lastovica A J, Elisha B G
Department of Medical Microbiology, University of Cape Town, Cape Town, South Africa.
J Clin Microbiol. 2001 Oct;39(10):3684-9. doi: 10.1128/JCM.39.10.3684-3689.2001.
A 1.6-kb DNA fragment isolated from a Campylobacter concisus genomic library gave C. concisus-specific restriction fragment length patterns when it was used as a probe in hybridization studies. All of the strains tested, including type strains and clinical isolates, contained a 0.5-kb HindIII fragment that hybridized to the probe. DNA sequencing of the 1.6-kb fragment identified three open reading frames (ORFs). One of the ORFs encodes the carboxy terminus of GyrB, and the translational products of ORF2 and ORF3 showed similarity to hypothetical proteins, previously identified in Campylobacter jejuni. DNA-DNA hybridization studies with a fragment internal to ORF3 showed that this sequence was responsible for the signal observed with the 0.5-kb HindIII fragment. A rapid PCR assay was developed and evaluated. Primers that annealed to the extremities of the 1.6-kb fragment were used to obtain an amplicon of the correct size from both reference and clinical strains of C. concisus.
从简明弯曲杆菌基因组文库中分离出的一个1.6kb DNA片段,在杂交研究中用作探针时,产生了简明弯曲杆菌特异性的限制性片段长度模式。所有测试菌株,包括模式菌株和临床分离株,都含有一个与该探针杂交的0.5kb HindIII片段。对该1.6kb片段进行DNA测序,鉴定出三个开放阅读框(ORF)。其中一个ORF编码GyrB的羧基末端,ORF2和ORF3的翻译产物与空肠弯曲杆菌中先前鉴定的假设蛋白具有相似性。用ORF3内部的一个片段进行DNA-DNA杂交研究表明,该序列是0.5kb HindIII片段所观察到信号的原因。开发并评估了一种快速PCR检测方法。使用与1.6kb片段末端退火的引物,从简明弯曲杆菌的参考菌株和临床菌株中均获得了正确大小的扩增子。