Elviss Nicola C, Lawson Andrew J, Owen Robert J
Helicobacter Reference Unit, Laboratory of Enteric Pathogens, Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK.
Int J Antimicrob Agents. 2004 Apr;23(4):349-55. doi: 10.1016/j.ijantimicag.2003.09.015.
Helicobacter pylori clarithromycin (Cla) resistance dramatically reduces efficacy of eradication therapy. In this study, 3'-mismatched reverse primer PCR (3M-PCR), real-time PCR (LightCycler), and PCR-RFLP assays were investigated to determine their sensitivity for detecting clarithromycin resistance associated with 23S rDNA mutations (A2142G, A2142C, and A2143G). For 84.8% (123/145) of isolates, the same allelic type was detected by each method although methods differed in efficiency of detecting mutations in cultures either containing mixtures of two alleles (24 isolates), or that were dual allelic variants (two isolates). The novel 3M-PCR assay format was the most sensitive, detecting all alleles at > or =0.02 ng/microl in DNA mixtures, and thus provides more precise information to guide clinical management of patients at risk of treatment failure.
幽门螺杆菌对克拉霉素（Cla）的耐药性会显著降低根除治疗的疗效。在本研究中，对3'-错配反向引物PCR（3M-PCR）、实时PCR（LightCycler）和PCR-RFLP分析方法进行了研究，以确定它们检测与23S rDNA突变（A2142G、A2142C和A2143G）相关的克拉霉素耐药性的敏感性。对于84.8%（123/145）的分离株，尽管不同方法检测含有两个等位基因混合物的培养物（24株分离株）或双等位基因变体（2株分离株）中的突变的效率有所不同，但每种方法检测到的等位基因类型相同。新型3M-PCR分析方法最为敏感，能在DNA混合物中检测到浓度≥0.02 ng/μl的所有等位基因，从而为指导有治疗失败风险患者的临床管理提供更精确的信息。
