Gramley W A, Asghar A, Frierson H F, Powell S M
Departments of Medicine, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA.
J Clin Microbiol. 1999 Jul;37(7):2236-40. doi: 10.1128/JCM.37.7.2236-2240.1999.
Stool, gastric biopsy, and serum samples were collected from 22 subjects. DNA from stool was extracted, amplified, and hybridized with primers specific for the 16S rRNA gene of Helicobacter pylori. DNA from gastric biopsy specimens was analyzed similarly for comparison. Universal primers were used to confirm successful extraction of DNA from samples. Histologic, serologic, and DNA analyses were scored in a blinded fashion. Universal primer amplification verified successful DNA extraction from all stool and gastric tissue specimens. The gastric tissue DNA assay was positive for H. pylori in 11 of the 22 subjects, correlating completely with histologic and serologic results. Stool DNA was positive for H. pylori by our molecular assay in 8 of these 11 H. pylori-positive subjects. All subjects who were negative by histologic, serologic, and gastric tissue DNA analyses were also negative by stool DNA analysis. Compared to histology, serology, and gastric tissue DNA analyses, the sensitivity of our stool DNA assay was 73%, with a specificity of 100%.
从22名受试者中收集粪便、胃活检组织和血清样本。提取粪便中的DNA,进行扩增,并与幽门螺杆菌16S rRNA基因的特异性引物杂交。对胃活检标本的DNA进行类似分析以作比较。使用通用引物确认从样本中成功提取了DNA。组织学、血清学和DNA分析采用盲法评分。通用引物扩增证实从所有粪便和胃组织标本中成功提取了DNA。在22名受试者中,11名的胃组织DNA检测显示幽门螺杆菌呈阳性,与组织学和血清学结果完全相符。在这11名幽门螺杆菌阳性受试者中,8名受试者的粪便DNA经我们的分子检测呈幽门螺杆菌阳性。所有组织学、血清学和胃组织DNA分析均为阴性的受试者,其粪便DNA分析也为阴性。与组织学、血清学和胃组织DNA分析相比,我们的粪便DNA检测的敏感性为73%,特异性为100%。