Rapid Detection, by PCR and Reverse Hybridization, of Mutations in the Helicobacter pylori 23S rRNA Gene, Associated with Macrolide Resistance |
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| Authors: | Leen-Jan van Doorn, Yvette J. Debets-Ossenkopp, Armelle Marais, Ricardo Sanna, Francis M graud, Johannes G. Kusters, Wim G. V. Quint |
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| Affiliation: | Delft Diagnostic Laboratory, Delft, The Netherlands. L.J.van.Doorn@ddl.nl |
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| Abstract: | 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. |
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