PCR-RFLP法检测骨钙素等位基因

Osteocalcin genotypes detected by polymerase chain reaction-restriction fragment length po- lymorphism

摘要: 目的: 为骨质疏松症的多基因综合研究提供一种简便、实用、准确的骨钙素(osteocalcin,OC)等位基因检测方法。方法: 采用聚合酶链反应(PCR)特异性扩增OC基因序列,扩增产物用限制性内切酶HindⅢ酶切,经聚丙烯酰胺凝胶电泳(PAGE)酸性银染后,观察酶切位点的限制酶片段长度多态性(RFLP)图谱。结果: 应用PCR RFLP法检测了77名健康汉族人OC基因型,hh、Hh、HH基因型分布频率分别为6 4%(49例)、32%(2 5例)、4%(3例)。结论: 该方法简便、准确、重复性好,适用于一般实验室及流行病学调查。

Abstract: Objective: To establish a simple,accurate and practical method for Osteocalcin （OC） genotype investigation Methods: OC gene was amplified by polymerase chain reaction（PCR） PCR products were digested with HindⅢ and subjected to PAGE and stained by silver nitrate The patterns of restriction fragment length polymorphism （RFLP） of OC gene were distinguished Results: The frequencies of genotype hh,Hh,HH were 64%,32% and 4% respectively in 77 healthy individuals Conclusion: PCR RFLP method is considered a simple and practical technique for obtaining accurate results in this research This method is suitable for routine laboratories and epidemiology studies on OC genotype determination