鹿胎DNA检测试剂盒的研制与评价  

Development and Evaluation of DNA Detection Kit for Fetus Cervi

作　　者：艾金霞[1] 李明成[1] 夏薇[1] 苑广信[2] 高峰[2] 高丽君[1] 张丽华 

机构地区：[1]北华大学医学检验学院,吉林吉林132013 [2]北华大学药学院,吉林吉林132013 [3]吉林市雷博科技有限公司,吉林吉林132013

出　　处：《中国药学杂志》2018年第4期306-311,共6页Chinese Pharmaceutical Journal

基　　金：吉林省科技发展计划资助项目(20160204004NY;20160307030YY;20170307001YY);吉林市科技创新发展计划资助项目(20162006)

摘　　要：目的研制一种鹿胎DNA检测试剂盒,对试剂盒的特异性、稳定性、敏感性和重复性进行评价,并对市售鹿胎药材进行质量考察。方法对经典鹿胎DNA提取及PCR鉴定方法进行改良,研制标准DNA检测试剂盒,用试剂盒对鹿胎正品、伪品进行DNA提取和PCR扩增,紫外分光光度法测纯度,对市售鹿胎样品进行验证。结果鹿胎DNA检测试剂盒反复冻融处理20次后依然有效,可于-20℃保存1年;样品DNA原液稀释200倍仍可检测;22个鹿胎样品中15个为正品,7个为不合格品。试剂盒特异性为100%;批内、批间变异系数分别为2.32%和2.56%。结论鹿胎DNA检测试剂盒的核酸提取量及纯度满足鉴定需求,且特异性好、灵敏度高、稳定性强,适用于鹿胎与伪品的快速检测。OBJECTIVE To develop a DNA detection kit for identification of Fetus Cervi, evaluate the quality indexes including specificity, stability, sensitivity and repeatability, and inspect the qualities of commercial Fetus Cervi samples. METHODS The Fetus Cervi DNA test kit was developed and modified using the classical DNA extraction and PCR identification method. The genomic DNAs of Fetus Cervi and counterfeit goods were extracted by the kit and PCR technique was performed to identify the authenticity. The purity of DNA was detected by UV spectrophotometer. Finally, commercially available Fetus Cervi samples were verified. RESULTS The kit was proved to be effective after 20 times of repeated frozen-thawing and it could be stored at - 20 ℃ for 1 year. The DNA could be detected when the primary solution was diluted by 200 times. The specificity test confirmed that the 15 samples were authentic Fetus Cervi, and 7 were counterfeit. The specificity of the kit was 100%. The repetitive experiments showed that tile average intra-assay CV% and inter-assay CV% of the kit were 2.32% and 2. 56% , respectively. CONCLUSION The amount and purity of the nucleic acids extracted by the DNA detection kit can meet the requirement for identification of Fetus Cervi, and the kit has good specificity, high sensitivity and good stability, so it is suitable for the rapid detection of Fetus Cervi.

关 键 词：鹿胎 DNA提取 DNA检测试剂盒 性能评价 

分 类 号：R284[医药卫生—中药学]