Development of a highly sensitive method for detection of clarithromycin-resistant Helicobacter pylori from human feces.
Gastric infection of clarithromycin (CAM)-resistant Helicobacter pylori is one of the major causes of failure to eradicate this organism. A noninvasive and useful method for the detection of CAM-resistant H. pylori from human feces by restriction fragment length polymorphism (RFLP)-nested polymerase chain reaction (PCR) targeting the mutation of the 23S rRNA gene that confers CAM-resistance in H. pylori was developed in this study. Our nested PCR method detected DNA of H. pylori in feces with high sensitivity and specificity compared with both an enzyme-linked immunoadsorbent assay (ELISA) of H. pylori in feces and the isolation of H. pylori from gastric biopsy. Furthermore, the results of mutation analysis of the H. pylori 23S rRNA gene amplified from feces completely correlated with both that of the H. pylori 23S rRNA gene amplified from the isolates of gastric biopsy and the susceptibility of H. pylori isolates to CAM. Therefore, our results show that this RFLP/nested PCR method is useful for the accurate diagnosis of CAM-resistant H. pylori infection from feces.
Rimbara
Emiko
E
Department of Microbiology, School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan.
Noguchi
Norihisa
N
Yamaguchi
Tai
T
Narui
Koji
K
Kawai
Takashi
T
Sasatsu
Masanori
M
D017291:Clarithromycin / Q000494:pharmacology*; D004352:Drug Resistance, Microbial / Q000235:genetics / Q000502:physiology*; D005243:Feces / Q000382:microbiology*; D016481:Helicobacter Infections / Q000175:diagnosis / Q000382:microbiology; D016480:Helicobacter pylori / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012338:RNA, Ribosomal, 23S / Q000032:analysis* / Q000235:genetics; D012680:Sensitivity and Specificity
D012338:RNA, Ribosomal, 23S; D017291:Clarithromycin