Hoshina S, Kahn S M, Jiang W, Green P H, Neu H C, Chin N, Morotomi M, LoGerfo P, Weinstein I B
Comprehensive Cancer Center, Columbia University, College of Physicians and Surgeons, New York, NY 10032.
Diagn Microbiol Infect Dis. 1990 Nov-Dec;13(6):473-9. doi: 10.1016/0732-8893(90)90079-b.
Helicobacter pylori is an organism thought to play an important causative role in gastritis and peptic ulcer diseases. We have designed an RNA dot blot assay for the detection of H. pylori, using as probe a synthetic oligonucleotide complementary to its 16S rRNA. We have also used oligonucleotide primers, complementary to conserved sequences within bacterial ribosomal 16S genes, to amplify a H. pylori ribosomal 16S DNA fragment via the polymerase chain reaction (PCR). After determining the DNA sequence of this amplified H. pylori fragment, primers were designed for specific PCR amplification of H. pylori ribosomal 16S DNA sequences. Samples from clinical endoscopic biopsies were PCR amplified with universal 16S ribosomal primers to detect the presence of bacteria and with H. pylori-specific primers to uniquely detect H. pylori. Finally, by comparing the H. pylori-specific PCR assay to commonly used diagnostic tests, we demonstrate that the molecular technique of PCR amplification shows promising applications for the clinical detection of H. pylori.
幽门螺杆菌是一种被认为在胃炎和消化性溃疡疾病中起重要致病作用的生物体。我们设计了一种RNA斑点印迹分析法来检测幽门螺杆菌，使用与它的16S rRNA互补的合成寡核苷酸作为探针。我们还使用了与细菌核糖体16S基因内保守序列互补的寡核苷酸引物，通过聚合酶链反应（PCR）扩增幽门螺杆菌核糖体16S DNA片段。在确定了这个扩增的幽门螺杆菌片段的DNA序列后，设计引物用于特异性PCR扩增幽门螺杆菌核糖体16S DNA序列。临床内镜活检样本用通用的16S核糖体引物进行PCR扩增以检测细菌的存在，并用幽门螺杆菌特异性引物进行PCR扩增以特异性检测幽门螺杆菌。最后，通过将幽门螺杆菌特异性PCR检测方法与常用诊断测试进行比较，我们证明PCR扩增的分子技术在幽门螺杆菌的临床检测中显示出有前景的应用。
