title: Enzymic colorimetry-based DNA chip: a rapid and accurate assay for detecting mutations for clarithromycin resistance in the 23S rRNA gene of Helicobacter pylori.
abstract: Macrolide drugs, such as clarithromycin (CAM), are a key component of many combination therapies used to eradicate Helicobacter pylori. However, resistance to CAM is increasing in H. pylori and is becoming a serious problem in H. pylori eradication therapy. CAM resistance in H. pylori is mostly due to point mutations (A2142G/C, A2143G) in the peptidyltransferase-encoding region of the 23S rRNA gene. In this study an enzymic colorimetry-based DNA chip was developed to analyse single-nucleotide polymorphisms of the 23S rRNA gene to determine the prevalence of mutations in CAM-related resistance in H. pylori-positive patients. The results of the colorimetric DNA chip were confirmed by direct DNA sequencing. In 63 samples, the incidence of the A2143G mutation was 17.46 % (11/63). The results of the colorimetric DNA chip were concordant with DNA sequencing in 96.83 % of results (61/63). The colorimetric DNA chip could detect wild-type and mutant signals at every site, even at a DNA concentration of 1.53 x 10(2) copies microl(-1). Thus, the colorimetric DNA chip is a reliable assay for rapid and accurate detection of mutations in the 23S rRNA gene of H. pylori that lead to CAM-related resistance, directly from gastric tissues.
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Shi-Hai
authors.initials: SH
authors.lastname: Xuan
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Yu-Gui
authors.initials: YG
authors.lastname: Zhou
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Bo
authors.initials: B
authors.lastname: Shao
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Ya-Lin
authors.initials: YL
authors.lastname: Cui
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Hong-Bo
authors.initials: HB
authors.lastname: Yin
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Song
authors.affiliation: Center of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001, PR China.
authors.forename: Hui
authors.initials: H
authors.lastname: Cong
authors.affiliation: Biochip Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, PR China.
authors.forename: Feng-Xiang
authors.initials: FX
authors.lastname: Jing
authors.affiliation: Biochip Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, PR China.
authors.forename: Qing-Hui
authors.initials: QH
authors.lastname: Jin
authors.affiliation: Center of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001, PR China.
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Wang
authors.affiliation: Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000328:Adult; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D017291:Clarithromycin / Q000494:pharmacology*; D003124:Colorimetry / Q000379:methods; D004269:DNA, Bacterial / Q000032:analysis; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D020459:Genes, rRNA; D016480:Helicobacter pylori / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D006801:Humans; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010458:Peptidyl Transferases / Q000235:genetics*; D017354:Point Mutation; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors; D055815:Young Adult
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D012338:RNA, Ribosomal, 23S; D010458:Peptidyl Transferases; D017291:Clarithromycin