Enzymic colorimetry-based DNA chip: a rapid and accurate assay for detecting mutations for clarithromycin resistance in the 23S rRNA gene of Helicobacter pylori.
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.
Xuan
Shi-Hai
SH
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Zhou
Yu-Gui
YG
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Shao
Bo
B
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Cui
Ya-Lin
YL
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Li
Jian
J
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Yin
Hong-Bo
HB
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Song
Xiao-Ping
XP
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Cong
Hui
H
Center of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001, PR China.
Jing
Feng-Xiang
FX
Biochip Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, PR China.
Jin
Qing-Hui
QH
Biochip Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, PR China.
Wang
Hui-Min
HM
Center of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001, PR China.
Zhou
Jie
J
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
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
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D012338:RNA, Ribosomal, 23S; D010458:Peptidyl Transferases; D017291:Clarithromycin