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Clarithromycin resistance and point mutations in the 23S rRNA gene in Helicobacter pylori isolates from Malaysia
Author(s) -
HO SinLoong,
TAN Eng Lai,
SAM Chook Koon,
GOH KheanLee
Publication year - 2010
Publication title -
journal of digestive diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.684
H-Index - 51
eISSN - 1751-2980
pISSN - 1751-2972
DOI - 10.1111/j.1751-2980.2010.00423.x
Subject(s) - 23s ribosomal rna , clarithromycin , helicobacter pylori , restriction fragment length polymorphism , point mutation , restriction enzyme , amplicon , biology , microbiology and biotechnology , polymerase chain reaction , drug resistance , gene , mutation , genetics , ribosome , rna
OBJECTIVE: To determine the prevalence of primary clarithromycin resistance amongst Helicobacter pylori (H. pylori) strains in Malaysian patients with gastroduodenal diseases, by using restriction fragment length polymorphism (RFLP) in domain V of 23S rRNA. METHODS: Gastric biopsies were obtained from H. pylori positive patients undergoing gastroscopy. DNA extraction was followed by PCR amplification using the primers Hp23‐1 and Hp23‐2 flanking a region of 425bp within the bacterial 23S rRNA peptidyltranferase (Hp23S fragment). Analysis of the 23S rRNA gene mutations is based on the generation of restriction sites for two restriction enzymes: Bbs I and Bsa I, which correspond to the base substitutions characteristic of clarithromycin resistance from A to G at positions 2142 and 2143, respectively. RESULTS: Gastric biopsy samples were obtained from 107 patients. A fragment of size 425bp corresponding to that expected from amplification of domain V of 23S rRNA was PCR‐amplified from only 105 samples. The amplicon was subsequently subjected to restriction by Bbs I and Bsa I. Only 1 sample (0.95%) had the Bbs I mutation (base substitution at A2142G) and 2 samples (1.90%) the Bsa I mutation (base substitution at A2143G). Thus 3 of 105 (2.9%) samples harbored clarithromycin resistant strains. CONCLUSION: In our experience, PCR‐RFLP is a rapid and precise method to detect the resistance of H. pylori to clarithromycin. Using this method, a low prevalence of clarithromycin resistance was detected in our local Malaysian strains. This augurs well for the continued use of clarithromycin as a first line drug in the treatment and eradication of H. pylori infection.