Rapid detection, by PCR and reverse hybridization, of mutations in the Helicobacter pylori 23S rRNA gene, associated with macrolide resistance.
A PCR-based reverse hybridization system (research prototype kit INNO-LiPA for H. pylori resistance) was developed and evaluated for simultaneous detection of 23S ribosomal DNA point mutations, associated with macrolide resistance in Helicobacter pylori. Fifty-seven H. pylori strains (51 natural, 6 laboratory-derived artificial, 52 resistant, and 5 susceptible strains) were tested by PCR-LiPA (detecting mutations A2115-->G, G2141-->A, A2142-->G, A2142-->C, A2143-->G, A2143-->C, and A2143-->T), DNA sequencing, restriction fragment length polymorphism, and/or hybridization to oligonucleotide probes. Results were highly concordant, but PCR-LiPA appears to be more sensitive for the simultaneous detection of multiple mutants.
van Doorn
L J
LJ
Delft Diagnostic Laboratory, Delft, The Netherlands. L.J.van.Doorn@ddl.nl
Debets-Ossenkopp
Y J
YJ
Marais
A
A
Sanna
R
R
Mégraud
F
F
Kusters
J G
JG
Quint
W G
WG
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004352:Drug Resistance, Microbial; D016480:Helicobacter pylori / Q000187:drug effects* / Q000235:genetics; D018942:Macrolides; D009154:Mutation; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012338:RNA, Ribosomal, 23S / Q000235:genetics*
D000900:Anti-Bacterial Agents; D018942:Macrolides; D012338:RNA, Ribosomal, 23S