摘要
为了有效地检测荷斯坦奶牛凝血因子Ⅺ缺乏症,试验以制备的纯合突变、野生型基因为模板,优化PCR反应体系,制备异源双链体系,建立多聚酶链反应-变性高效液相色谱(PCR-DHPLC)的检测方法,确定最佳的检测条件,并用建立的方法对120个样本进行检测,再与传统的方法进行比较和验证。结果表明:所建立的PCR-DHPLC方法可以用于检测牛凝血因子Ⅺ缺乏症,与传统方法的检测结果一致。说明PCR-DHPLC方法是一种高通量、准确性高、特异性好的检测方法。
To effectively detect the coagulation factor XI deficiency in Holstein cows, the mutant type and wild type genes were used as the tem- plates, and the PCR reaction system was optimized. And then the hetero - duplex DNA system was prepared. The polymerase chain reaction - denaturing high - performance liquid chromatography（ PCR - DHPLC） was established to confirm the best examination condition. The 120 sam- pies were investigated by PCR - DHPLC, and then were compared and validated using traditional method. The results showed that the estab- lished PCR - DHPLC method could be used for the detection of the coagulation factor XI deficiency in Hdstein cows,and the test results using the established PCR - DHPLC method were the same as those using traditional method. It is concluded that PCR - DHPLC is a detection meth- od with high throughout, high accuracy, and good specificity.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2013年第11期102-104,191,共4页
Heilongjiang Animal Science And veterinary Medicine
基金
质检公益性行业科研专项(201210122)
关键词
凝血因子XI
携带者
变性高效液相色谱(DHPLC)
coagulation factor XI
carrier
denaturing high- performance liquid chromatography（ DHPLC ）
