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J Med Microbiol ; 58(Pt 11): 1443-1448, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19628643

RESUMO

Macrolide drugs, such as clarithromycin (CAM), are a key component of many combination therapies used to eradicate Helicobacter pylori. However, resistance to CAM is increasing in H. pylori and is becoming a serious problem in H. pylori eradication therapy. CAM resistance in H. pylori is mostly due to point mutations (A2142G/C, A2143G) in the peptidyltransferase-encoding region of the 23S rRNA gene. In this study an enzymic colorimetry-based DNA chip was developed to analyse single-nucleotide polymorphisms of the 23S rRNA gene to determine the prevalence of mutations in CAM-related resistance in H. pylori-positive patients. The results of the colorimetric DNA chip were confirmed by direct DNA sequencing. In 63 samples, the incidence of the A2143G mutation was 17.46 % (11/63). The results of the colorimetric DNA chip were concordant with DNA sequencing in 96.83 % of results (61/63). The colorimetric DNA chip could detect wild-type and mutant signals at every site, even at a DNA concentration of 1.53 x 10(2) copies microl(-1). Thus, the colorimetric DNA chip is a reliable assay for rapid and accurate detection of mutations in the 23S rRNA gene of H. pylori that lead to CAM-related resistance, directly from gastric tissues.


Assuntos
Antibacterianos/farmacologia , Claritromicina/farmacologia , Farmacorresistência Bacteriana/genética , Helicobacter pylori/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Peptidil Transferases/genética , RNA Ribossômico 23S/genética , Adulto , Colorimetria/métodos , DNA Bacteriano/análise , Genes de RNAr , Helicobacter pylori/enzimologia , Helicobacter pylori/genética , Humanos , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Mutação Puntual , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA , Fatores de Tempo , Adulto Jovem
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