Detection of Helicobacter pylori in gastric mucosa of patients with gastroduodenal diseases by PCR-restriction analysis using the RNA polymerase gene (rpoB).
A novel PCR restriction analysis method using the RNA polymerase beta-subunit- coding gene (rpoB) was employed to both detect and identify Helicobacter pylori in biopsy specimens and culture isolates. The rpoB DNAs (458 bp) were specifically amplified by PCR with the Helicobacter-specific primers (HF and HR). Based on the determined rpoB sequences of the culture isolates, an H. pylori-specific restriction site, Tru9I, was found. H. pylori can be identified by observing two discernible DNA fragments (288 and 138 bp) after Tru9I digestion and agarose gel electrophoresis. The rpoB PCR and subsequent restriction analysis (PRA) enabled the specific detection and identification of H. pylori in biopsy specimens from patients with gastroduodenal diseases. The rpoB PRA conferred a compatible or a slightly higher positive rate (53.7%) than did the Campylobacter-like organism (CLO) test (50.4%) and glmM PCR (48.8%), suggesting that it is useful for diagnosing an H. pylori infection without culture in the clinical laboratory.
Lim
Chang-Young
CY
Department of Internal Medicine, Dankook University College of Medicine, Cheonan, 330-715, Korea.
Lee
Keun-Hwa
KH
Cho
Myung-Je
MJ
Chang
Myung-Woong
MW
Kim
Seok-Yong
SY
Myong
Na-Hye
NH
Lee
Woo-Kon
WK
Rhee
Kwang-Ho
KH
Kook
Yoon-Hoh
YH
D015373:Bacterial Typing Techniques; D001706:Biopsy; D003470:Culture Media; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004378:Duodenal Diseases / Q000175:diagnosis / Q000382:microbiology; D005753:Gastric Mucosa / Q000382:microbiology*; D016481:Helicobacter Infections / Q000175:diagnosis / Q000382:microbiology; D016480:Helicobacter pylori / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013272:Stomach Diseases / Q000175:diagnosis / Q000382:microbiology
D003470:Culture Media; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit