NEW METHOD FOR QUANTITATIVE MEASUREMENT OF HELICOBACTER PYLORI IN GASTRIC MUCUS AND GASTRIC MUCOSA.

定量测量胃粘液和胃粘膜中幽门螺杆菌的新方法。

• 批准号: 06670535
• 负责人: KANEKO Eizo
• 金额: $ 1.34万
• 依托单位: HAMAMATSU UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670535/
• 关键词: HELICOBACTER PYRORI; COMPETITIVE-PCR; PEPTIC ULCER; ヘリコパクターピロリ; competitive PCR法; Helicobacter pylori; プロトンポンプインヒビター; H.pyloriの除菌; pepsinogen I; II

Objectives : Helicobacter pylori (Hp) is closely related to upper gastrointestinal diseases. Establishment of a precise diagnostic device for Hp is of great value. The aim of this study was establishment of a method for quantitatively detecting the bacteria. Methods : We developed a new methods for quantitative detection of Hp by competitive polymerase chain reaction (cPCR) using various known amounts of synthetic DNA fragments containing the same primer template sequences as the target competing for primer binding and amplification. The quantity of Hp in gastric mucus and gastric tissue was measured by this method. The results obtained with this method in gastric mucus were compared with results of histology, rapid urease test (RUT), bacterial culture, and ^<13>C-urea breath test (^<13>C-UBT). The distribution of Hp in the gastric mucosa was also studied quantitatively in the conection with gastric function. Results : Relatively large quantity (10^5-10^9/mucus 100mul) of Hp were found … More by cPCR in gastric mucus when results of histology, RUT or bacterial culture were positive. Even when results of histology, RUT or bacterial culture for Hp were negative, a considerable quantity of Hp in gastric mucus (10^5-10^<8/> mucus 100mul) was often found by cPCR.The quantity of Hp in gastric mucus is significantly correlated with results of ^<13>C-UBT (p<0.0005). We also confirmed that this method was able to distinguish the true negative from the false negative due to incomplete PCR,and the true positive from the false positive due to contamination. As for the distribution of Hp, quantity of Hp in corpus was larger than that in antrum in patients with GU.Quantity of Hp in corpus was smaller than that of antrum in patients with DU.The pepsinogen I/II ratio in the serum significantly increases by eradication of Hp and this change of the ratio was confirmed to be an useful index for the eradicaiton. Conclusions : These results suggest that the cPCR method is very useful for precise and quantitative evaluation of Hp infection in the stomach. Less

目的：幽门螺杆菌（Hp）与上消化道疾病密切相关。建立精确的Hp诊断装置具有重要的价值。本研究的目的是建立一种定量检测细菌的方法。方法：采用竞争性聚合酶链反应（cPCR）定量检测Hp的新方法，利用不同数量的合成DNA片段，这些DNA片段含有相同的引物模板序列作为目标，竞争引物结合和扩增。用该方法测定胃粘液和胃组织中Hp的含量。将本法在胃粘液中得到的结果与组织学、快速脲酶试验（RUT）、细菌培养和^<13> c -尿素呼气试验（^<13>C-UBT）的结果进行比较。定量研究Hp在胃黏膜的分布与胃功能的关系。结果：胃液中Hp数量较多（10^5 ~ 10^9/ 100mul），组织学、RUT或细菌培养结果均为阳性时，cPCR检测较多。即使组织学、RUT或细菌培养结果为阴性，cPCR也经常在胃粘液中发现相当数量的Hp（10^5-10^<8/>粘液100mul）。胃粘液中Hp的数量与^<13>C-UBT结果显著相关（p<0.0005）。我们还证实，该方法能够区分因PCR不完全导致的真阴性和假阴性，以及因污染导致的真阳性和假阳性。在Hp的分布上，GU患者的Hp在体中的数量大于在胃窦中的数量。脓毒症患者体中Hp的数量少于胃窦。血清胃蛋白酶原I/II比值因Hp的根除而显著升高，该比值的变化被证实为根除Hp的有用指标。结论：cPCR方法可用于胃Hp感染的精确定量评价。少

项目成果

Furuta T: "Helicoba cfer pylori-preseut studus in Japan and a newer diagnasfic method applying compatitive PCR" Chinese J. Gastroeufeol. Hepafol.5(in press). (1996)

Furuta T：“日本幽门螺杆菌预研究和应用相容性 PCR 的新诊断方法”中国 J. Gastroeufeol。

二見 肇: "Competitive PCR法を用いた消化性潰瘍患者における粘膜組織内Helicobacter pyloviの定量" 日本消化器内視鏡学会雑誌. 37. 114- (1995)

Hajime Futami：“使用竞争性PCR方法定量消化性溃疡患者粘膜组织中的幽门螺杆菌”日本胃肠内窥镜学会杂志37. 114-（1995）。

T.Furuta: "Eftect of Laniscprizale with or without Hijh-Pole Aivoxicillin on Helscobcaten pylori in peptic ulcer patients" Suprelsion of Acid Secvetion by Lansoprazale and Ies Effect or lt pylor. 2. 73-75 (1994)

T.Furuta：“Laniscprizale 联合或不联合 Hijh-Pole 艾伏西林对消化性溃疡患者幽门螺杆菌的影响”兰索拉扎抑制胃酸分泌和幽门螺杆菌效应。

T.Furuta: "Quantificontion of Helicobacar pylni in gastsic inncus 7 conpetitive PCR wig synthetic DNA fragments" 10th Wovcl Cogrecs of Grastroencerology. 148- (1994)

T.Furuta：“幽门螺杆菌在胃内的定量 7 竞争性 PCR 假发合成 DNA 片段”第 10 届 Wovcl Cogrecs of Gratroncerology。

Furuta T et al: "Percentage changes in serum pepsinogen are useful as indexes of eradication of Helicobacter pylori" Amer J Gastroenterl. (submitted).

Furuta T 等人：“血清胃蛋白酶原的百分比变化可用作根除幽门螺杆菌的指标”Amer J Gastroenterl。