Comparing Multiplex PCR and Rapid Urease Test in the Detection of H. pylori in Patients on Proton Pump Inhibitors.

Background. This study was conducted to assess the diagnostic value of a multiplex PCR assay to detect H. pylori infection and to further evaluate the negative results from the CLOtest on patients with and without PPI treatment. Methods. This study is a retrospective cohort that included 457 patients with symptoms of dyspepsia, who underwent upper endoscopy at Evanston and Glenbrook Northshore Hospital from June 2003 to October 2007. A total of 556 samples were reported with some patients having more than one test over the time period. The CLOtest was performed first on the gastric specimen and from that specimen, the DNA was isolated and the one-step multiplex PCR was performed. Results. By M-PCR testing, H. pylori was detected in 143 (52.2%) of 274 cases in the control group and 130 (46.1%) of 282 cases in patients on PPI treatment (P = 0.1746). The CLOtest detected the presence of H. pylori in 4 (1.4%) of 282 cases from the same group receiving PPI treatment and 29 (10.6%) of 274 cases from the group not taking a PPI (P ≤ 0.0001). Conclusion. Our PCR is sensitive enough to detect the presence of H. pylori despite being on PPI treatment.

Detection of Helicobacter pylori gastritis by PCR: correlation with inflammation scores and immunohistochemical and CLOtest findings.

We developed a polymerase chain reaction (PCR) assay to detect Helicobacter pylori in gastric and/or gastroesophageal biopsy specimens of adults with dyspepsia, compared the method with immunohistochemical analysis and CLOtest (Ballard Medical Products, Draper, UT), and correlated the results of each test with the histologic features of infection. H pylori was identified in 36 (60%) of 60 patients irrespective of biopsy site and testing method. In the gastric biopsy specimens, PCR detected H pylori in 29 (52%) of 56 cases, including 11 (100%) of 11 immunohistochemically and/or CLOtest-positive cases. PCR-positive gastric biopsy specimens correlated with a higher average cumulative inflammatory score compared with PCR-negative specimens (P = .001). In gastroesophageal biopsy specimens, PCR detected H pylori in 15 (34%) of 44 cases, including 1 (20%) of 5 immunohistochemically positive specimens. PCR-positive gastroesophageal junction biopsy specimens did not correlate with a higher average cumulative inflammatory score. Overall, PCR detected an additional 23 cases negative by immunohistochemical analysis and/or CLOtest. This PCR assay identified a significant number of H pylori infections that would not be detected by immunohistochemical analysis and/or CLOtest.

Inhibition of bacterial adherence on the surface of stents and bacterial growth in bile by bismuth dimercaprol.

Bacterial infection and biofilm formation on the surface of biliary stents is believed to be one of the main factors in stent occlusion. This study explored the role of the new reagent, bismuth dimercaprol, in preventing bacterial adherence and bacterial biofilm formation on the surface of biliary stents. Sterile porcine bile preparations, infected separately with Escherichia coli, Klebsiella pneumoniae, Enterobacter, and Enterococcus, were used as the perfusion media in an in vitro perfusion system. The bacterial growth in the media and the bacterial adherence on the surface of stents were tested when different concentrations of bismuth dimercaprol were used in the perfusion media. BisBAL (5 microM) did not inhibit the growth of any of the tested bacterial species. It did, however, significantly decrease the amount of bacteria adhering to the surface of stents for all bacterial strains except Escherichia coli. Bismuth dimercaprol (20 microM) significantly inhibited the growth of Escherichia coli, Klebsiella pneumoniae, and Enterobacter and, thereby, significantly decreased the amount of these bacteria adhering to the surface of stents. The unique bactericidal and anitbiofilm activities of bismuth thiols might contribute to delaying the process of biliary stent occlusion if the effective concentrations of bismuth thiols could be delivered to the target sites. The feasibility of this application of bismuth thiols deserves further investigation.

Bile glycoprotein mucin in sludge occluding biliary stent.

Endoscopic biliary stenting is a common treatment for the palliation of obstructive jaundice caused by inoperable malignant hepatobiliary tumors and benign strictures. The biliary stent, however, often becomes nonfunctional as a result of occlusion. In this study, we sought to confirm that bile glycoprotein mucin was a factor in stent occlusion and to investigate its possible role in biliary-stent blockage. The high-molecular-weight glycoprotein fraction was isolated from stent sludge with the use of gel filtration and a cesium chloride density gradient. This fraction was analyzed for amino-acid and carbohydrate compositions and was identified by means of immunoblotting with a specific monoclonal antibody against human gallbladder mucin. Furthermore, the distribution of bile glycoprotein mucin in stent sludge was immunologically demonstrated with fluorescent antibody, and the relationship between bile glycoprotein mucin and bacteria (demonstrated with DAPI stain) was observed. The high-molecular-weight glycoprotein extracts isolated from 11 patients' stent sludge showed strongly positive immunoreactivity with the monoclonal antibody against human gallbladder mucin. Immunofluorescence studies showed that very bright fluorescent signals of bile glycoprotein mucin often appeared on the surface of pigmented deposits, at the periphery of clumps of bacteria and along the inner wall of stents. In nonpigmented sludge, we noted fluorescent signals of bile glycoprotein mucin dispersed among clumps of bacteria. Bile glycoprotein mucin is a constituent of stent sludge. It may play a role in stent occlusion by affecting bacterial adherence to the stent surface or by promoting stent-sludge accumulation as one kind of cement among substances such as calcium bilirubinate and clumps of bacteria.

Evaluation of role of bile flow patterns in process of in vitro stent occlusion.

Stent placement above the sphincter of Oddi might have advantages over stent placement across the sphincter of Oddi in prolonging stent patency in the treatment for malignant obstructive jaundice. To evaluate the role of bile flow patterns corresponding to biliary stent positioning in the process of stent occlusion in an in vitro bile perfusion model, one group of polyethylene stents was perfused continuously and another group of stents was perfused with additional flushing three times a day, simulating gallbladder emptying. After 8 weeks, the flow rates through the perfused stents were measured for evaluating the extent of stent occlusion indirectly. The results showed that bile flow rate of stents with additional flushing was significantly higher than the continuously perfused stents (P < or = 0.01). It was demonstrated that after 18 hr of perfusion, additional flushing obviously decreased bacterial adherence to stent when compared to continuously perfused stents. In conclusion, flushing of bile may decrease the build-up of substance in vitro and thus improve stent flow rates, for which decreasing bacterial adherence to stents may be responsible.

Role of bile mucin in bacterial adherence to biliary stents.

Biliary stent placement is a well-established method of relieving obstructive jaundice. However, a frequent complication is occlusion of the stent caused by bacterial biofilm formation and sludge accumulation. In this study we investigated the possible effect of bile mucin on bacterial adherence to biliary stents at the initial stage of biofilm formation. By means of an in vitro bile-perfusion system, polyethylene stents were perfused with pig gallbladder bile infected with Escherichia coli. The concentrations of mucin in the pig bile were adjusted with purified mucin. The amount of bacteria adhering to the inner surface of the stents was measured and compared for stents perfused with bile containing various concentrations of mucin. Furthermore, we conditioned the stent inner surface with purified pig bile mucin and observed the effect of the conditioning on subsequent bacterial adherence. In addition, a common method for assaying bacterial adhesion with polystyrene microtiter plates was also used in this study. The results demonstrated that more bacteria adhered to the inner surface of stents perfused with bile containing 5 mg/mL mucin than of those perfused with bile containing 0.5 and 0 mg/mL mucin. Increased bacterial adherence was demonstrated on the stent surfaces conditioned with purified mucin compared with that seen on the nonconditioned stent surfaces. The optical densities indicating bacterial adhesion in the microtiter plate wells precoated with mucin were higher than those in non-coated plate wells. The in vitro results indicate that when a biliary stent is implanted in vivo, mucin in bile may condition the stent inner surface, modulate subsequent bacterial adherence to the surface, and participate in stent occlusion.