Capillary electropherograms for restriction fragment length polymorphism of Helicobacter pylori.
Rapid identification of Helicobacter pylori strains is of importance for diagnosis and then treatment of duodenal and gastric ulcers. We developed a CE approach for the analysis of RFLP of the PCR products of urease (UreAB) gene and flagellin A (FlaA) gene fragments. Prior to CE analysis, the 2.4-kbp UreAB and 1.5-kbp FlaA PCR products were digested with the restriction enzymes HaeIII and HhaI, respectively. The DNA fragments were then separated by CE in conjunction with laser-induced fluorescence detection using poly(ethylene oxide) in the presence of electroosmotic flow. The DNA fragments range in sizes 259-1831 bp and 12-827 bp for UreAB and FlaA restriction fragments, respectively. Of 27 samples, the CE approach provided five and ten different RFLP patterns of the HaeIII and HhaI digests. The RFLP of PCR products of the two genes allow great sensitivity of identification of H. pylori strains. When compared with slab gel electrophoresis, the present CE approach provides advantages of rapidity (within 6 min per run), simplicity, and automation. The preliminary results have shown great practicality of the CE approach for screening H. pylori strains.
Bair
Ming-Jong
MJ
Gastroenterology Division, Department of Internal Medicine, Mackay Memorial Hospital, Taitung Branch, Taiwan.
Chen
Chiu-Lin
CL
Chiang
Cheng-Kang
CK
Huang
Ming-Feng
MF
Hu
Cho-Chun
CC
Chang
Huan-Tsung
HT
D001426:Bacterial Proteins / Q000235:genetics; D004262:DNA Restriction Enzymes; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D005408:Flagellin / Q000235:genetics; D005798:Genes, Bacterial; D016481:Helicobacter Infections / Q000382:microbiology; D016480:Helicobacter pylori / Q000145:classification / Q000235:genetics*; D006801:Humans; D007834:Lasers; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D014510:Urease / Q000235:genetics
D001426:Bacterial Proteins; D005408:Flagellin; C067138:flaA protein, bacteria; D004262:DNA Restriction Enzymes; D014510:Urease