摘要
目的:针对正品鹿茸与近源种鹿茸药材饮片鉴别的难点,建立一种简便、快速、准确的近源种鹿茸药材分子鉴别方法。方法:根据鹿科鹿属种动物Cytb全序列比对分析,设计1对能准确鉴别正品鹿茸(包括梅花鹿和马鹿)和其他近源种鹿茸的引物,在此基础上建立鹿茸药材位点特异性PCR鉴别方法,并对影响PCR结果的主要因素退火温度、Taq用量、循环次数以及模板用量和重复性等进行方法学考察和优化。结果:在方法学考察的基础上,在25μL PCR反应体系,退火温度为65℃的条件下,能准确扩增出约323 bp大小的阳性DNA条带,经测序验证结果表明,系正品鹿茸原动物的Cytb基因序列片段,而伪品鹿茸未扩增出条带。结论:所建立的位点特异性PCR方法,能将正品鹿茸与其他近源种鹿茸药材鉴别开来,具有较高的特异性、重复性,可广泛应用于鹿类药材的鉴别。
Objective:To establish a convenient,quick and accurate molecular method for the identification of crude drugs of antlers due to the difficult discrimination between the genuine antler and its adulterants.Method: According to the alignment analysis of full length sequences of Cytb gene from closely relate species of Cervus,one pair of allele-specific diagnostic PCR primers was designed.Factors such as annealing temperature,dosage of polymerase,times of cycles and dosage of template DNA that influence the PCR...
出处
《中国中药杂志》
CAS
CSCD
北大核心
2009年第23期3013-3016,共4页
China Journal of Chinese Materia Medica
基金
<中国药典>2010版一部标准研究项目(YD-195-1)
关键词
位点特异性PCR
近源种鹿茸药材
鉴别
Allele-specific diagnostic PCR
antlers from closely relate species of Cervus
identification
