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

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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