Detection of Point Mutations Associated with Resistance of Helicobacter pylori to Clarithromycin by Hybridization in Liquid Phase

摘要: When the standard procedure for determining antibiotic susceptibility of bacteria is used, results are delayed, especially that grow slowly, such as Helicobacter pylori. Treatment this bacterium may involve clarithromycin, a compound which resistance has been associated with point mutations on 23S rRNA gene. This currently found in organisms isolated from 0 to 15% patients and jeopardizes success treatment. We have designed test involving amplification colorimetric hybridization liquid phase detect mutation at molecular level. First, four reference strains, including wild type three strains A2143C, A2143G, A2144G, were used optimize method. Amplification was carried out primers previously published. The amplified products added probe-coated microtiter wells. A DNA enzyme immunoassay hybrids. optimal conditions defined each probe. Nineteen H. pylori resistant clarithromycin 22 susceptible according phenotypic data submitted restriction BsaI BbsI, part gene sequenced order determine involved strains. new assay showed complete correlation methods, except one strain. Cross-hybridizations well application reaction other did not lead optical densities higher than cutoff values chosen receiving operating characteristic curve. method can be easily standardized gives result within day. Its directly biopsy specimens or infected gastric juice planned future.