高分辨熔解曲线非标记探针基因分型技术检测恶性疟原虫青蒿素耐药相关K13基因突变方法学的建立  

Establishment of a methodology for detecting artemisinin-Resistant K13 gene mutationsin Plasmodium falciparum using High-Resolution Melting curve unlabeled probe genotyping

作　　者：陈伟忠 CHEN Weizhong(Department of Clinical Laboratory,Chaozhou People′s Hospital,Chaozhou,Guangdong 521000,China)

机构地区：[1]广东省潮州市人民医院检验科,广东潮州521000

出　　处：《检验医学与临床》2021年第13期1908-1910,1914,共4页Laboratory Medicine and Clinic

基　　金：广东省医学科学技术研究基金项目(A2019198)。

摘　　要：目的建立一种通过高分辨熔解曲线(HRM)-非标记探针基因分型分析方法,检测恶性疟原虫青蒿素耐药相关K13基因C580Y突变位点。方法通过构建野生型和C580Y突变型质粒,设计相应引物、探针,应用HRM-非标记探针技术检测C580Y突变。结果成功检测出标本中含有C580Y突变的基因型及野生基因型。检测野生型和突变型的探针峰的温度之差为5℃。结论HRM-非标记探针技术是一种能够快速、准确、廉价、反应通量大的检测方法,适用于疫区恶性疟原虫青蒿素耐药相关K13基因突变的大批量检测。Objective To establish a High-Resolution Melting(HRM)analysis with an Unlabeled Probe for artemisinin resistance related mutation site C580Y of K13 gene in Plasmodium falciparum.Methods The wild-type and C580Y mutant plasmids were constructed,and the corresponding primer probes were designed.The C580Y mutation was detected by HRM analysis with an Unlabeled Probe.Results Genotypes containing C580Y mutation and wild genotypes were successfully detected in the samples.The difference in temperature between the wild-type and mutant probe peaks was 5℃.Conclusion The high resolution melting analysis with unlabeled probes technique is a fast,accurate,inexpensive,and high-throughput assay,suitable for the large-scale detection of K13 gene mutations associated with Plasmodium falciparum artemisinin resistance in endemic areas.

关 键 词：恶性疟原虫 HRM 基因突变 

分 类 号：Q343.12[生物学—遗传学]