Xuan Shi-Hai, Zhou Yu-Gui, Shao Bo, Cui Ya-Lin, Li Jian, Yin Hong-Bo, Song Xiao-Ping, Cong Hui, Jing Feng-Xiang, Jin Qing-Hui, Wang Hui-Min, Zhou Jie
Department of Clinical Laboratory, The People's Hospital of Dongtai City, Dongtai 224200, PR China.
Center of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001, PR China.
J Med Microbiol. 2009 Nov;58(Pt 11):1443-1448. doi: 10.1099/jmm.0.010785-0. Epub 2009 Jul 23.
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.
大环内酯类药物，如克拉霉素（CAM），是用于根除幽门螺杆菌的多种联合疗法的关键组成部分。然而，幽门螺杆菌对CAM的耐药性正在增加，并且在幽门螺杆菌根除治疗中成为一个严重问题。幽门螺杆菌对CAM的耐药性主要归因于23S rRNA基因肽基转移酶编码区域的点突变（A2142G/C、A2143G）。在本研究中，开发了一种基于酶比色法的DNA芯片，用于分析23S rRNA基因的单核苷酸多态性，以确定幽门螺杆菌阳性患者中与CAM相关耐药性的突变发生率。比色DNA芯片的结果通过直接DNA测序得到证实。在63个样本中，A2143G突变的发生率为17.46%（11/63）。比色DNA芯片的结果与DNA测序结果在96.83%的结果（61/63）中一致。比色DNA芯片甚至在DNA浓度为1.53×10²拷贝微升⁻¹时，也能在每个位点检测到野生型和突变型信号。因此，比色DNA芯片是一种可靠的检测方法，可直接从胃组织中快速准确地检测幽门螺杆菌23S rRNA基因中导致与CAM相关耐药性的突变。
