应用非标记探针高分辨率熔解曲线技术检测人巨细胞病毒UL97基因突变

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (3): 322-326.

应用非标记探针高分辨率熔解曲线技术检测人巨细胞病毒UL97基因突变

赵晓涛¹,夏长胜¹,孙媛媛¹,王启¹,于波²,邵勇张正¹

收稿日期:2012-01-04 修回日期:2012-01-06 出版日期:2012-03-05 发布日期:2012-02-27
通讯作者: 赵晓涛 E-mail:zhxt510@hotmail.com

Development of high-resolution melting analysis with unlabeled probe for the detection of human cytomegalovirus UL97 mutation

Received:2012-01-04 Revised:2012-01-06 Online:2012-03-05 Published:2012-02-27

摘要/Abstract

摘要： 目的建立非标记探针高分辨率熔解曲线分析技术（HRMA）检测人巨细胞病毒（HCMV）UL97基因突变。方法构建HCMV UL97野生型以及H520Q突变质粒，非标记探针HRMA分型技术检测H520Q突变。结果直接从PCR产物得到的熔解曲线峰型无法区分野生型及H520Q突变型，应用非标记探针HRM A技术可以明显增加实验的敏感性及准确性，可以成功区分从野生型到含不同量的H520Q突变的基因型。结论非标记探针HRMA 技术是一个敏感、经济的检测H520Q突变的基因分型方法。

关键词: 人巨细胞病毒（HCMV）, UL97, 高分辨率熔解曲线技术 (HRMA)

Abstract: Objective To develop a high resolution melting analysis (HRMA)-based genotypic testing for the detection of UL97 mutations of human cytomegalovirus (HCMV). Methods Wild-type UL97 and H520Q mutant were constructed. HRMA with unlabeled probe were used as genotyping method for the detection of H520Q mutation. Results The melting peaks directly from the PCR products were not able to distinguish wild-type from H520Q mutation, and the sensitivity and accuracy of HRMA were dramatically improved using unlabeled probe. HRMA with unlabeled probe successfully distinguished different contents of H520Q from wild-type. Conclusions HRMA with unlabeled probe proves to be a sensitive and cost-effective genotyping method for the detection of H520Q mutation.

Key words: human cytomegalovirus（HCMV）, UL97, high resolution melting analysis (HRMA)

赵晓涛夏长胜孙媛媛王启于波邵勇张正. 应用非标记探针高分辨率熔解曲线技术检测人巨细胞病毒UL97基因突变[J]. 基础医学与临床, 2012, 32(3): 322-326.