Co-detection of Helicobacter pylori and of its resistance to clarithromycin by PCR.
Our aim was to develop a rapid molecular test based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and making it possible to detect Helicobacter pylori directly from gastric biopsy samples, and to test its susceptibility to clarithromycin. A 629-bp fragment of the 23S rRNA gene of H. pylori was amplified by PCR and the mutations responsible for clarithromycin resistance were detected with Bsa1 and Bbs1 restriction endonucleases. Thirty-five gastric samples were tested in parallel by standard microbiologic methods (culture and clarithromycin susceptibility testing with E-test strips) and by PCR-RFLP. The 10 culture-negative samples were also PCR-negative. Sixteen out of the 25 culture-positive samples (64%) were PCR-positive. RFLP analysis could be done in 12 cases and the results were in agreement with those of the E-test: susceptibility in five cases, resistance in seven (six A2144G mutations and one A2143G mutation).
Sevin
E
E
Service de Bactériologie-Virologie-Hygiène, Hôpital Henri Mondor, Créteil, France.
Lamarque
D
D
Delchier
J C
JC
Soussy
C J
CJ
Tankovic
J
J
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015373:Bacterial Typing Techniques; D017291:Clarithromycin / Q000494:pharmacology*; D004275:DNA, Ribosomal / Q000032:analysis; D004352:Drug Resistance, Microbial / Q000235:genetics; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000145:classification / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification*; D006801:Humans; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011706:Pyloric Antrum / Q000382:microbiology*; D012338:RNA, Ribosomal, 23S / Q000235:genetics
D000900:Anti-Bacterial Agents; D004275:DNA, Ribosomal; D012338:RNA, Ribosomal, 23S; D017291:Clarithromycin