Detection of Helicobacter Species from Korean Patients with Gallstones Using Polymerase Chain Reactions

Lee, Dong Ki; Baik, Soon Ku; Kwon, Sang Ok; Kim, Hyun Soo; Kim, Jong Bae; Kim, Geun Hee; Kim, Hong; Ha, Soo Hyun

Abstract

BACKGROUND/AIMS: Helicobacter pylori and bile-resistant Helicobacter spp. Which inhabit in animals were reported to be detected in human samples and to be associated with the hepatobiliary tract diseases of such as cholelithiasis and cholecystitis. Because the rate of H. pylori infection is above 90% in Korea, it is important to investigate the prevalence of Helicobacter spp. In biliary tract. We investigated the prevalence of Helicobacter spp. In gastric biopsy, gastric juice, gallbladder tissue, bile juice, and gallstone samples from Korean patients with cholelithiasis.
METHODS
Clinical biliary specimens of gallstone (n=51), bile juice (n=43), and gallbladder tissue (n=29) were collected from 68 Korean patients with symptomatic gallstone, using aseptic technique during cholecystectomy. Concomitantly, gastric mucosa (n=60) and gastric juice samples (n=42) were obtained before cholecystectomy. DNA from these specimens was extracted and amplified with the Helicobacter genus-specific and species-specific novel primers for 16S rRNA gene. For confirmation of the H. pylori, H. hepaticus, and H. felis specific PCR-amplified DNA, Southern blot hybridization was performed.
RESULTS
The gallbladder tissue specimens were positive for H. pylori in 4, for H. hepaticus in 1, and for H. felis in 2 samples. The bile juice specimens were positive for H. pylori in 12, for H. hepaticus in 1, and for H. pullorum in 2 samples. The gallstone specimens were positive only for H. pylori in 11 samples. Interestingly, 12 gastric biopsy, 5 gastric juice, and 1 bile juice sample which was positive in H. hepaticus-specific PCR were also positive in PCR with H. pylori-specific primers. This result was completely correlated with the result of Southern blot hybridization using corresponding internal probes of hpp225 and hhp961, respectively.
CONCLUSIONS
The results of this study may imply that H. pylori and H. hepaticus can be co-infected in patients with biliary diseases.