摘要
针对虎线粒体细胞色素b(Cyt b)基因设计特异性引物,建立并优化SYBR GreenⅠ实时荧光PCR检测鉴定体系。利用该体系分别对从虎(东北虎、孟加拉虎、印支虎)和狮、豹、熊等13种非虎哺乳动物的肌肉、血液、毛发和肉骨粉中提取的DNA进行鉴定分析。结果表明:虎血液、毛发和肌肉DNA样本均出现特异性的实时扩增曲线,平均熔解温度在(83.8±0.1)℃。狮、豹、熊等13种非虎哺乳动物DNA样本的检测结果均为阴性。检测灵敏度可达到在骨粉中检出质量分数为0.05%的虎源性成分。SYBR GreenⅠ实时定量PCR方法具有方便、经济、灵敏度高和重复性好等特性,可用于虎制品的检测鉴定。
This paper developed a convenient SYBR Green Ⅰ-based real-time PCR assay which allows identification of Panthera tigris-specific mitochondrial DNA in meat,hair,blood and bone meal.The specificity of this method was confirmed by blood,hair and meat samples from Panthera tigris altaica,P.t.tigris and P.t.corbetti as well as blood and bone meal samples from 13 non-P.tigris mammalian species.Results showed that DNA samples of blood,hair and meat from Panthera t.altaica,P.t.tigris and P.t.corbetti exhibited specifically real-time amplification curves,and the average melting temperature was（83.8±0.1）℃.The detections of DNA samples from 13 non-P.tigris mammalian species were all negative.This method could detect the presence of P.tigris material in bone meal mixture even when the concentration of P.tigris-derived meat was reduced to 0.05%.Furthermore,it did not appear to be affected by prolonged heat treatment.This method can be developed for food detection,fur identification and traditional Chinese drug identification.
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2010年第10期82-84,共3页
Journal of Northeast Forestry University
基金
公益性行业科研专项(2007GYJ037)
