Marshall D G, Coleman D C, Sullivan D J, Xia H, O'Moráin C A, Smyth C J
Moyne Institute of Preventive Medicine, Department of Microbiology, School of Dental Science, Trinity College, University of Dublin, Republic of Ireland.
J Appl Bacteriol. 1996 Nov;81(5):509-17. doi: 10.1111/j.1365-2672.1996.tb03540.x.
The ability of oligonucleotide probes containing short repetitive sequence motifs to differentiate between isolates of Helicobacter pylori was investigated. Genomic DNA preparations from H. pylori were digested with the restriction enzyme HindIII, electrophoresed in agarose gels and transferred to nylon filters. Five separate oligonucleotide probes were tested for hybridization sequentially to fingerprint the digested DNA from a panel of 29 clinical isolates and one type strain of H. pylori, and their relative discriminatory abilities were assessed. Four probes, (GACA)4, (GT)8, (GTG)5 and (GGAT)4, were each shown to yield highly informative hybridization band profiles allowing differentiation of H. pylori isolates. The DNA fingerprints of individual isolates obtained with each probe were distinct and reproducible. Direct comparison with ribotyping revealed that oligonucleotide fingerprinting had far superior discriminatory power. Computer-assisted similarity analysis of (GGAT)4-generated hybridization profiles of pairwise combinations of H. pylori isolates revealed that there was no correlation between ribotype and oligonucleotide fingerprint patterns. The results of this study demonstrate that oligonucleotide probes containing microsatellite sequences provide a new and powerful tool for isolate discrimination of H. pylori.
研究了含有短重复序列基序的寡核苷酸探针区分幽门螺杆菌分离株的能力。用限制性内切酶HindIII消化幽门螺杆菌的基因组DNA制剂，在琼脂糖凝胶中进行电泳并转移到尼龙滤膜上。依次测试了五个单独的寡核苷酸探针用于杂交，以对来自29株临床分离株和一株幽门螺杆菌标准菌株的消化DNA进行指纹分析，并评估它们的相对鉴别能力。四个探针，(GACA)4、(GT)8、(GTG)5和(GGAT)4，均显示产生了高度信息丰富的杂交带谱，能够区分幽门螺杆菌分离株。用每个探针获得的各个分离株的DNA指纹是独特且可重复的。与核糖体分型的直接比较表明，寡核苷酸指纹分析具有远为优越的鉴别能力。对幽门螺杆菌分离株两两组合的(GGAT)4产生的杂交图谱进行计算机辅助相似性分析表明，核糖体分型与寡核苷酸指纹图谱之间没有相关性。本研究结果表明，含有微卫星序列的寡核苷酸探针为幽门螺杆菌分离株的鉴别提供了一种新的强大工具。
