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題 名 | Genotyping of Helicobacter Pylori Isolates by Random Amplified Polymorphic DNA and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism=AP-PCR與PCR-RFLP對幽門旋曲桿菌分型 |
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作 者 | 林振文; 王淑芬; 鄭庚申; | 書刊名 | 中臺灣醫學科學雜誌 |
卷 期 | 3:2 1998.06[民87.06] |
頁 次 | 頁101-107 |
分類號 | 414.83 |
關鍵詞 | 幽門旋曲桿菌; 聚合酵素鏈反應; 限制酵素切割片斷分析圖; 隨意引子放大片段分析圖; PCR; RFLP; RAPD; Helicobacter pylori; Polymerase chain reaction; Restriction fragment length polymorphism; Randomly amplified polymorphic DNA; |
語 文 | 英文(English) |
中文摘要 | 幽門旋曲桿菌被認為菌株多樣化可能與幽門旋曲桿多種臨床病徵有關。幽門旋曲桿菌urease基因之多樣化可提供區別菌株之處。本論文中,我們從二十二位病人分離出幽門旋曲桿菌並抽取染色體DNA,再調查RFLP對PCR放大╱820-bp ureC基因之分型分析,並與RAPD對菌株染色體分析比較。HindIII及EcoRI切割22位病患消化道切片檢體分離菌株之PCR放大820-bp ureC基因之分型分析只有1種RFLP型式,而Sau3A分型分析有6種不同RFLP型式。Sau3A之第1種RFLP型式有9株,為22分離菌株的40.9%。我們以RAPD對22分離菌株之染色體之分型分析可區分為21種RAPD型式。Sau3A之RFLP分型分析圖比RAPD對染色體之分型分析圖簡單、容易判讀。RFLP對PCR產物之分型能力好且可將分離菌分群。比較幽門旋曲桿菌基因之多樣化,RFLP及RAPD可得到快速及可信賴的結果來了解幽門旋曲桿菌之傳染途徑及致病機轉。 |
英文摘要 | Helicobacter pylori is thought to show strain diversity. The differences in the pathology of H. pylori may have to do with this diversity. We saw the use of polymerase chain reaction (PCR) to amplify the ureC in H. pylori and the examination of restriction fragment length polymorphism (RELP) within the ureC region as a means to type clinical isolates of H. pylori, where point mutations in the sequence diversity of the ureC gene can be found. Likewise, we saw the use of randomly amplified polymorphic DNA(RAPD) to type genotypic variation in the gene order on physical maps, mosaicism in conserved gene, non-conserved genes and extragenetic elements. In this study, H. pylori genomic DNA was prepared from strains isolated from 22 unrelated patients and examined by PCR-based typing of H. pylori isolates using RAPD analysis of genomic DNA and RFLP analysis of ureC gene with HindIII, EcoRI and Sau3A. From the restriction enzyme digestion of an 820-bp fragment from within the ureC gene in H. pylori strains isolated from the 22 patients, we found only one pattern with HindIII or EcoRI, but 6 different patterns with Sau3A. Nine strains (40.9%) were grouped RFLP pattern 1 with Sau3A digestion of ure C region. We found 21 different RAPD patterns of genomic DNA. The H. pylori RFLP patterns of ureC gene gave much simpler profiles than those produced by RAPD analysis of gemonic DNA. In summary, H. pylori isolates have great genetic diversity among strains, which can be exploited to type isolates. RAPD on the whole bacterial genome can accurately distinguish between isolates. RFLP analysis with Sau3A of the ureC gene also had a good discriminatory power and group isolates into clusters. Comparing the genetic diversity of H. pylori isolated from different patients, these methods yield fast and reliable results on the transmission and pathogenicity of H. pylori. |
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