The Study On The Effect Of Helicobacter Pylori In The Formation Of Pigment Stone By NP-PCR Technology

Objective: To explore the effect of bacteria, especially to helicobacter pylori in the formation of pigment stone with negative bacterial culture by analyzing bacteria DNA in the hepatolithiasis and intrahepatic tissue. To study the relation of infection of helicobacter pylori and the content of pigment by checking the content of pigment in the intrahepatic pigment stone.Methods: Collecting pigment stone of patients who suffered from hepatolithiasis in aseptic conditions, 48 cases entered the study, and we used nested primers polymerase chain reaction (NP-PCR) technique to amplify bacterial gene fragments extracted from intrahepatic pigment stone. In addition, comparative 16s ribosomal RNA sequence analysis was used for elucidation of bacterial identification. Enrlich diazo reaction test was used to measure the content of pigment in stones. Meanwhile, we collected 30 cases of biliary tract tissues of patients who suffered from hepatolithiasis which were supposed research group and 20 cases of normal biliary tract tissues which were supposed control group in aseptic conditions. Then, the technology of agarose gel electrophoresis was used to analyze DNA fragment and DNA sequence was determined.Results: Bacterial DNA was found in the stones of 42 patients (87.5%) . Helicobacter pylori DNA was found in the stones of 12 patients (25%). E.coli-related DNA fragments were found in 10 cases (20.83%) . Propionibacteria type DNA was found in 8 cases (16.67%) . While, pseudomonas aeruginosa gene fragments were found in 5 cases (10.42%) . A more heterogenous sequence collection was found in 7 patients (14.58%) . In addition, in 6 cases of stones which was passive in the inspection of bacteria DNA, the content of pigment was 83. 4 ±5.3. In 42 cases of stones which was positive in the inspection of bacteria DNA, the content of pigment was 152.0±12.8 . There were obvious differences (P<0. 01). And the content of pigment was 250.0 ± 12.2 in stones which was positive in the inspection of Helicobacter pylori DNA, the content of pigment was 163. 0 ± 5. 3 in stones which was positive in the inspection of E.coli DNA, the content of pigment was 172. 0±7.5in stones which was positive in the inspection of Propionibacteria DNA, the content of pigment was 169. 0 ± 7. 2 in stones which was positive in the inspection of pseudomonas aeruginosa DNA. There were magnificent differences as far as Helicobacter pylori was concerned compared with other kinds (P<0. 05) . Besides them, the positive rate of Helicobacter pylori DNA was 40% in biliary tract tissues of hepatolithiasis, while the positive rate of Helicobacter pylori DNA was 10% in normal biliary tract tissues . It goes without saying that there were remarkable differences between two groups (P<0. 01).Conclusions: It goes without saying that pigment stone could be assigned to the multiple bacterial infections. Moreover, there are obvious differences between the content of pigment and bacterial infections. As soon as helicobacter pylori is concerned, it is of much importance to the nuclei of origin of intrahepatic pigment stone.