The recognition of the role of Helicobacter pylori in gastric diseases has led to the widespread use of antibiotics in the eradication of this pathogen. The most advocated therapy, triple therapy, often includes clarithromycin. It is well known that clarithromycin resistance is one of the major causes of eradication failure. The development of a rapid noninvasive technique that could easily be performed on fecal samples and that could also provide information about the antibiotic resistance of this microorganism is therefore advisable. Previous findings have demonstrated that clarithromycin resistance is due to a single point mutation in the 23S rRNA. All the mutations described have been associated with specific restriction sites, namely BsaI (A2143G), MboII (A2142C/G), and HhaI (T2717C). On this basis we have developed a new method, a seminested PCR, allowing screening for clarithromycin resistance of H. pylori directly on stool samples. This method furnished a 783-bp fragment of the 23S rRNA, which was subsequently digested by MboII, BsaI, and HhaI, in order to identify single point mutations associated with clarithromycin resistance. Of a total of 283 stool samples examined, 125 were H. pylori positive and two of them were shown to contain clarithromycin-resistant strains due to the presence of a mutation at position 2717, whereas no PCR products contained mutations at position 2142 or 2143. In order to evaluate the reliability of the new system, we compared the results of restriction analysis of the PCR products with the MICs shown by the H. pylori isolates by culturing gastric biopsies from the same patients.

Fontana, C., Favaro, M., Pietroiusti, A., Pistoia, E.s., Galante, A., Favalli, C. (2003). Detection of clarithromycin-resistant Helicobacter pylori in stool samples, 41(8), 3636-3640 [10.1128/JCM.41.8.3636-3640.2003].

Detection of clarithromycin-resistant Helicobacter pylori in stool samples

FONTANA, CARLA;FAVARO, MARCO;PIETROIUSTI, ANTONIO;PISTOIA, ENRICO SALVATORE;GALANTE, ALBERTO;FAVALLI, CARTESIO
2003-01-01

Abstract

The recognition of the role of Helicobacter pylori in gastric diseases has led to the widespread use of antibiotics in the eradication of this pathogen. The most advocated therapy, triple therapy, often includes clarithromycin. It is well known that clarithromycin resistance is one of the major causes of eradication failure. The development of a rapid noninvasive technique that could easily be performed on fecal samples and that could also provide information about the antibiotic resistance of this microorganism is therefore advisable. Previous findings have demonstrated that clarithromycin resistance is due to a single point mutation in the 23S rRNA. All the mutations described have been associated with specific restriction sites, namely BsaI (A2143G), MboII (A2142C/G), and HhaI (T2717C). On this basis we have developed a new method, a seminested PCR, allowing screening for clarithromycin resistance of H. pylori directly on stool samples. This method furnished a 783-bp fragment of the 23S rRNA, which was subsequently digested by MboII, BsaI, and HhaI, in order to identify single point mutations associated with clarithromycin resistance. Of a total of 283 stool samples examined, 125 were H. pylori positive and two of them were shown to contain clarithromycin-resistant strains due to the presence of a mutation at position 2717, whereas no PCR products contained mutations at position 2142 or 2143. In order to evaluate the reliability of the new system, we compared the results of restriction analysis of the PCR products with the MICs shown by the H. pylori isolates by culturing gastric biopsies from the same patients.
2003
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore MED/44 - MEDICINA DEL LAVORO
English
Con Impact Factor ISI
clarithromycin; ribosome RNA; virus RNA; adult; aged; antibiotic resistance; article; bacterial infection; bacterial strain; bacterium isolate; controlled study; disease association; feces; Helicobacter pylori; human; human tissue; major clinical study; minimum inhibitory concentration; nucleotide sequence; point mutation; polymerase chain reaction; preschool child; priority journal; reliability; restriction mapping; sampling; screening test; stomach biopsy; stomach disease; tissue culture; treatment failure; virus mutation; Anti-Bacterial Agents; Base Pairing; Base Sequence; Clarithromycin; DNA Primers; Drug Resistance, Bacterial; Feces; Gene Amplification; Helicobacter Infections; Helicobacter pylori; Humans; Polymerase Chain Reaction; Restriction Mapping; Sensitivity and Specificity; Bacteria (microorganisms); Helicobacter; Helicobacter pylori
Fontana, C., Favaro, M., Pietroiusti, A., Pistoia, E.s., Galante, A., Favalli, C. (2003). Detection of clarithromycin-resistant Helicobacter pylori in stool samples, 41(8), 3636-3640 [10.1128/JCM.41.8.3636-3640.2003].
Fontana, C; Favaro, M; Pietroiusti, A; Pistoia, Es; Galante, A; Favalli, C
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/50119
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