欢迎访问《中国临床药理学与治疗学》杂志官方网站,今天是

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (6): 647-651.

• 临床药理学 • 上一篇    下一篇

焦磷酸测序技术检测LAMA5 rs944895基因多态性方法的建立

贾政军1, 王华1, 彭向京1, 黄定梅1, 王果2   

  1. 1湖南省妇幼保健院遗传室,长沙 410008,湖南;
    2中南大学临床药理研究所,长沙 410078,湖南
  • 收稿日期:2011-05-06 修回日期:2011-05-20 发布日期:2011-07-25
  • 通讯作者: 王果,男,博士后,副教授,研究方向:遗传药理与临床药理学。 Tel: 0731-84805380 E-mail: wangguo30@yahoo.com.cn
  • 作者简介:贾政军,女,硕士,副主任医师,研究方向:临床遗传学与优生优育。 Tel: 13975867315 E-mail: jiazhengj@tom.com
  • 基金资助:
    国家自然科学基金项目(81072706);湖南省科技计划项目(2009JT3020);教育部博士点(新教师)基金项目(20090162120024);湖南省卫生厅科研基金

Development of pyrosequencing method for detection of LAMA5 rs944895

JIA Zheng-jun1, WANG Hua1, PENG Xiang-jing1, HUANG Ding-mei1, WANG Guo2   

  1. 1 The Maternal and Child Health Hospital of Hunan Province, Changsha 410008, Hunan, China;
    2 Institute of Clinical Pharmacology, Central South University, Changsha 410078, Hunan, China
  • Received:2011-05-06 Revised:2011-05-20 Published:2011-07-25

摘要: 目的: 建立LAMA5 rs944895基因多态性分析的焦磷酸测序方法,并分析该多态性在中国健康人群中的分布频率。方法: 制备201例健康人外周血gDNA,应用PyroMark ID焦磷酸测序仪进行LAMA5 rs944895基因多态位点的焦磷酸测序,并经过重复性试验和毛细管电泳测序验证该方法的准确性。结果: 建立了LAMA5 rs944895基因多态性分析的焦磷酸测序新方法,检出率100%,重复性试验和毛细管电泳测序验证准确可靠。201例健康人中A等位基因频率为 72.4%,G为 27.6%;AA、AG、GG等位基因型频率分别为 53.2%、38.3%和 8.5%,符合Hardy-Weinberg平衡(P=0.55)。结论: LAMA5 rs944895基因多态性分析的焦磷酸测序新方法具有快速、准确和高通量的优点,适合于在科研和临床应用中推广。

关键词: 焦磷酸测序, 多态性, LAMA5, 序列分析

Abstract: AIM: To establish a pyrosequencing based method for detection LAMA5 rs944895 polymorphism and to determine the frequency of this polymorphism in healthy Chinese. METHODS: After preparation of gDNA from blood of 201 subjects, LAMA5 rs944895 target fragments were amplified by PCR, LAMA5 rs944895 polymorphism was detected on PyroMark ID by pyrosequencing technology. The reliability of pyrosequencing method was validated by repeat tests and Sanger sequencing. RESULTS: We established a new pyrosequencing method to detect the LAMA5 rs944895 polymorphism in healthy Chinese. The detection rate and repetition rate were both 100%. The frequencies of A allele and G allele were 72.4% and 27.6%, respectively. And the frequencies of AA, AG and GG genotype were 53.2%, 38.3% and 8.5%, respectively. The genotype frequencies match the Hardy-Weinberg equilibrium (P=0.55). CONCLUSION: This pyrosequencing assay to detect the LAMA5 rs944895 polymorphism is proved to be a rapid, accurate and high-throughput alternative to conventional methods, and it can be a preferred option in research and clinical application.

Key words: Pyrosequencing, Polymorphism, LAMA5, Sequencing

中图分类号: