Majewski S I, Goodwin C S
Department of Microbiology, Royal Perth Hospital, Western Australia.
J Infect Dis. 1988 Mar;157(3):465-71. doi: 10.1093/infdis/157.3.465.
We developed a rapid extraction method to analyze the chromosomal DNA of 84 isolates of Campylobacter pylori (formerly Campylobacter pyloridis) from 70 individuals. Only three of the nine endonucleases tested gave satisfactory digestions: HindIII, EcoRI, and SacI. The latter two produced mostly larger bands, whereas HindIII produced smaller bands, which allowed clearer comparisons between isolates. The isolates from 69 Australian subjects and one from England each had a unique profile. Isolates from a husband and wife were different, as were those from a brother and sister. In pairs of isolates from 11 individuals, the second isolate was markedly different in six subjects. The profile changes were not associated with changes in antibiotic susceptibility or with loss of catalase or urease activity, which can occur during storage of C. pylori. These restriction endonuclease profiles suggest considerable subspecies variation in C. pylori. Plasmid bands were found in undigested DNA from 40 of the 84 C. pylori isolates.
我们开发了一种快速提取方法,用于分析来自70个人的84株幽门螺杆菌(原幽门弯曲菌)的染色体DNA。在所测试的9种核酸内切酶中,只有3种酶切效果令人满意:HindIII、EcoRI和SacI。后两种酶产生的条带大多较大,而HindIII产生的条带较小,这使得分离株之间的比较更加清晰。来自69名澳大利亚受试者和1名来自英国受试者的分离株各有独特的图谱。夫妻的分离株不同,兄弟姐妹的分离株也不同。在来自11个人的分离株对中,6名受试者的第二个分离株明显不同。图谱变化与抗生素敏感性变化或在幽门螺杆菌储存期间可能发生的过氧化氢酶或尿素酶活性丧失无关。这些限制性内切酶图谱表明幽门螺杆菌存在相当大的亚种差异。在84株幽门螺杆菌分离株中,有40株未消化的DNA中发现了质粒条带。