Premium
Capillary electropherograms for restriction fragment length polymorphism of Helicobacter Pylori
Author(s) -
Bair MingJong,
Chen ChiuLin,
Chiang ChengKang,
Huang MingFeng,
Hu ChoChun,
Chang HuanTsung
Publication year - 2008
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200700911
Subject(s) - haeiii , restriction enzyme , restriction fragment length polymorphism , flagellin , microbiology and biotechnology , capillary electrophoresis , biology , restriction fragment , helicobacter pylori , terminal restriction fragment length polymorphism , dna sequencer , polymerase chain reaction , chemistry , dna , gene , chromatography , genetics
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 Hae III and Hha I, 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 Hae III and Hha I 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.