Wada S, Matsuda M, Kikuchi M, Kodama T, Takei I, Ogawa S, Takahashi S, Shingaki M, Itoh T
Department of Biosciences, College of Environmental Health, Azabu University, Sagamihara, Japan.
Cytobios. 1994;80(321):109-16.
The genotype of genome DNA from seventeen clinical isolates of Helicobacter pylori by pulsed-field gel electrophoresis (PFGE) and arbitrarily primed polymerase chain reaction (AP-PCR) was determined. Three restriction enzymes, Apa I, Kpn I and Not I, were found to produce distributions of DNA fragments which were useful for analysis of the chromosome-sized DNA from H. pylori NCTC11637T by PFGE. Many of the isolates of H. pylori could not be genotyped by PFGE after digestion reaction with Apa I, Kpn I, and Not I. When AP-PCR with a ten-nucleotide primer was performed using the chromosomal genomic DNA from the isolates as templates, fifteen distinctly different profiles were obtained from the seventeen isolates. Thus, genotyping of the isolates was possible where PFGE profiles had not previously been informative. The results suggest that AP-PCR is more suitable for genotyping of clinical isolates of H. pylori in Japan than PFGE.
通过脉冲场凝胶电泳（PFGE）和任意引物聚合酶链反应（AP-PCR）测定了17株幽门螺杆菌临床分离株基因组DNA的基因型。发现三种限制性内切酶Apa I、Kpn I和Not I产生的DNA片段分布，可用于通过PFGE分析幽门螺杆菌NCTC11637T的染色体大小的DNA。许多幽门螺杆菌分离株在与Apa I、Kpn I和Not I进行消化反应后，无法通过PFGE进行基因分型。当使用分离株的染色体基因组DNA作为模板，用一个十核苷酸引物进行AP-PCR时，从17株分离株中获得了15种明显不同的图谱。因此，在PFGE图谱以前无信息的情况下，对分离株进行基因分型是可能的。结果表明，在日本，AP-PCR比PFGE更适合用于幽门螺杆菌临床分离株的基因分型。
