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J Med Microbiol 58 (2009), 1443-1448; DOI: 10.1099/jmm.0.010785-0
© 2009 Society for General Microbiology
ISSN 0022-2615

Enzymic colorimetry-based DNA chip: a rapid and accurate assay for detecting mutations for clarithromycin resistance in the 23S rRNA gene of Helicobacter pylori

Shi-Hai Xuan1, Yu-Gui Zhou1, Bo Shao1, Ya-Lin Cui1, Jian Li1, Hong-Bo Yin1, Xiao-Ping Song1, Hui Cong2, Feng-Xiang Jing3, Qing-Hui Jin3, Hui-Min Wang2 and Jie Zhou1

1 Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China

2 Center of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001, PR China

3 Biochip Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, PR China

Correspondence
Feng-Xiang Jing
jingfengx{at}163.com
Hui-Min Wang
zj6669{at}163.com

Received March 4, 2009
Accepted July 16, 2009

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 1.53x102 copies µl–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.






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