Li C, Ferguson D A, Ha T, Chi D S, Thomas E
Department of Internal Medicine (Gastroenterology), James H. Quillen College of Medicine, East Tennessee State University, Johnson City 37684.
J Clin Microbiol. 1993 Aug;31(8):2157-62. doi: 10.1128/jcm.31.8.2157-2162.1993.
HindIII-digested DNA fragments derived from an EcoRI-digested 6.5-kb fragment of chromosomal DNA prepared from Helicobacter pylori ATCC 43629 (type strain) were cloned into the pUC19 vector. A 0.86-kb insert was identified as a potential chromosomal DNA probe. The specificity of the probe was evaluated by testing 166 non-H. pylori bacterial strains representing 38 genera and 91 species which included aerobic, anaerobic, and microaerophilic flora of the upper and lower gastrointestinal tracts. None of the 166 non-H. pylori strains hybridized with this probe (100% specificity), and the sensitivity of this probe was also 100% when H. pylori isolates from 72 patients with gastritis and with the homologous ATCC type strain were tested by dot blot hybridization. The capability of this probe for differentiating between strains of H. pylori was evaluated by Southern blot hybridization of HaeIII-digested chromosomal DNA from 68 clinical isolates and the homologous ATCC type strain of H. pylori. Fifty-one unique hybridization patterns were seen among the 69 strains tested, demonstrating considerable genotypic variation among H. pylori clinical isolates. We propose that this probe would be of significant value for conducting epidemiologic studies.
从幽门螺杆菌ATCC 43629(模式菌株)制备的经EcoRI酶切的6.5 kb染色体DNA片段中获得的经HindIII酶切的DNA片段,被克隆到pUC19载体中。一个0.86 kb的插入片段被鉴定为潜在的染色体DNA探针。通过检测代表38个属和91个种的166株非幽门螺杆菌菌株来评估该探针的特异性,这些菌株包括上、下胃肠道的需氧菌、厌氧菌和微需氧菌菌群。166株非幽门螺杆菌菌株中没有一株与该探针杂交(特异性为100%),当通过斑点印迹杂交检测来自72例胃炎患者的幽门螺杆菌分离株和同源ATCC模式菌株时,该探针的灵敏度也为100%。通过对68株临床分离株和幽门螺杆菌同源ATCC模式菌株经HaeIII酶切的染色体DNA进行Southern印迹杂交,评估该探针区分幽门螺杆菌菌株的能力。在所检测的69株菌株中观察到51种独特的杂交模式,表明幽门螺杆菌临床分离株之间存在相当大的基因型变异。我们认为该探针对于进行流行病学研究具有重要价值。