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

目的：幽门螺杆菌(Hp)与上消化道疾病密切相关。建立一种准确的幽门螺杆菌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：我们建立了一种竞争聚合酶链式反应(CPCR)定量检测幽门螺杆菌的新方法，该方法以各种已知数量的含有相同模板序列的合成DNA片段为竞争目标。用该方法检测胃粘液和胃组织中Hp的含量。并与组织学、快速尿素酶试验(RUT)、细菌培养和C-尿素呼气试验(C-UBT)结果进行比较。结合胃功能，定量研究幽门螺杆菌在胃粘膜中的分布。结果：发现较多数量的…(10^5~10^9/黏液100mul)当组织学、RUT或细菌培养结果为阳性时，更多地采用CPCR方法检测胃粘液。即使在组织学、RUT或细菌培养阴性的情况下，cPCR也能在胃粘液中发现相当数量的Hp(105-10；8/&gt；100mul)。胃粘液中Hp的数量与C-UBT的结果显著相关(p&lt；0.0005)。我们还证实，该方法能够区分由于PCR不完全而导致的真阴性和假阴性，以及由于污染而导致的真阳性和假阳性。从Hp的分布来看，胃溃疡患者胃体Hp含量大于胃窦Hp含量，DU患者胃体Hp含量小于胃窦Hp含量。根除Hp后，血清胃蛋白酶原I/II比值显著升高，这一比值的变化可作为根除Hp的有用指标。结论：CPCR法可用于准确、定量地评价胃内Hp感染情况。较少

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