PCR-LDR-核酸试纸条检测法在结核分枝杆菌katG基因315位密码子突变检测中的应用

【作者】 贾子冬；

【导师】 朱金玲； 罗佳滨； 尤其敏； 胡林；

【作者基本信息】 佳木斯大学 ， 遗传学， 2011， 硕士

【摘要】 目的:改良优化一项新的分子生物学方法PCR-LDR-核酸试纸条检测法,并将其应用于结核分枝杆菌katG基因315位密码子单碱基突变的检测。为临床检测结核分枝杆菌耐药相关单碱基突变提供一种新方法,以指导临床用药。方法:设计与合成用于检测结核分枝杆菌katG基因315位密码子单碱基突变的引物和探针,通过LDR反应,使用核酸检测试纸条检测,并与PCR-RFLP法及直接测序法的检测结果相比较。结果:成功检测各单碱基突变类型及野生型,与PCR-RFLP法及直接测序法的检测结果呈高度一致性,符合率分别为97.5%和100%。结论:该方法可有效检测结核分枝杆菌katG基因315位密码子的单碱基突变,并为临床检测结核分枝杆菌耐药相关单碱基突变提供一种新的快速简便廉价的分子诊断方法。更多还原

【Abstract】 Objective:This study aims to meliorate and optimize a novel molecular diagnositic method(PLNA), prove the feasibility of this method for detecting the site specific mutations of katG gene condon 315 of Mycobacterium tuberculosis , and apply this method to the distruction of clinical drug use .Methods:Primers and probes were designed and synthesized for the detection of mutations in katG gene codon315 of Mycobacterium tuberculosis , performed the LDR reaction , and then detected use the nucleic acid detection strip , the results were compared with PCR-RFLP and direct sequencing .Results:The wildtype and mutants were successfully diagnosed , a high consistency was found compared with PCR-RFLP and direct sequencing ,the concordance rate was 97.5% and 100% respectively .Conclusion:This study provided a viable low-cost quick molecular diagnosis method for the clinical detection of mutations at katG gene codon 315 of Mycobacterium tuberculosis.更多还原