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Bacteriology

Rapid Detection of Point Mutations Conferring Resistance to Fluoroquinolone in gyrA of Helicobacter pylori by Allele-Specific PCR

Toshihiro Nishizawa, Hidekazu Suzuki, Ayako Umezawa, Hiroe Muraoka, Eisuke Iwasaki, Tatsuhiro Masaoka, Intetsu Kobayashi, Toshifumi Hibi
Toshihiro Nishizawa
1Department of Internal Medicine
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Hidekazu Suzuki
1Department of Internal Medicine
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  • For correspondence: hsuzuki@sc.itc.keio.ac.jp
Ayako Umezawa
2Center for Integrated Medical Research, Keio University School of Medicine
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Hiroe Muraoka
3Mitsubishi Kagaku Bio-Clinical Laboratories
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Eisuke Iwasaki
1Department of Internal Medicine
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Tatsuhiro Masaoka
4Department of Gastroenterology, Eiju Hospital, Tokyo, Japan
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Intetsu Kobayashi
3Mitsubishi Kagaku Bio-Clinical Laboratories
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Toshifumi Hibi
1Department of Internal Medicine
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DOI: 10.1128/JCM.01997-06
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    FIG. 1.

    Representative electrophoresis patterns of gyrA quinolone resistance-determining region (QRDR) PCR products and allele-specific PCR products amplified with mixed primers for determination of C261A, C261G, G271A, G271T and A272G mutant gyrA genotypes of H. pylori. The theoretical fragment sizes according to the gyrA sequence are mentioned in the text.

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  • TABLE 1.

    Oligonucleotide sequences of primers used in AS-PCR method to determine gyrA mutation of H. pylori at positions 261, 271, and 272

    Primer directionMutationPrimer namePrimer sequenceaPCR product size (bp)
    ForwardC261AF261A1CCCCCATGGCGAGAAaG262
    C261GF261G1CCCCCATGGCGAGAAgG262
    G271AF271A5GCGATAACGCGGTTTAGaA254
    F271A9GCGATAATGCGGTTTAGaA254
    G271TF271T9GGCGATAATGCGGTTAATtA255
    A272GF272G1GCGATAACGCGGTTTAGGgT254
    F272G9GCGATAATGCGGTTTAGGgT254
    ReversegyrA RGTTAGGCAGACGGCTTGGTARAATA
    • ↵ a The penultimate nucleotide to distinguish between wild-type and mutant sequences is shown in lowercase. The 1-bp mismatch at another nucleotide is underlined.

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Rapid Detection of Point Mutations Conferring Resistance to Fluoroquinolone in gyrA of Helicobacter pylori by Allele-Specific PCR
Toshihiro Nishizawa, Hidekazu Suzuki, Ayako Umezawa, Hiroe Muraoka, Eisuke Iwasaki, Tatsuhiro Masaoka, Intetsu Kobayashi, Toshifumi Hibi
Journal of Clinical Microbiology Feb 2007, 45 (2) 303-305; DOI: 10.1128/JCM.01997-06

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Rapid Detection of Point Mutations Conferring Resistance to Fluoroquinolone in gyrA of Helicobacter pylori by Allele-Specific PCR
Toshihiro Nishizawa, Hidekazu Suzuki, Ayako Umezawa, Hiroe Muraoka, Eisuke Iwasaki, Tatsuhiro Masaoka, Intetsu Kobayashi, Toshifumi Hibi
Journal of Clinical Microbiology Feb 2007, 45 (2) 303-305; DOI: 10.1128/JCM.01997-06
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KEYWORDS

Anti-Infective Agents
DNA Gyrase
Drug Resistance, Bacterial
fluoroquinolones
Helicobacter pylori
Point Mutation
polymerase chain reaction

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