摘要
[目的]建立荷斯坦牛凝血因子Ⅺ的PCR检测方法。[方法]参照GenBank上登录的牛FⅪ(FactorⅪ)基因序列,设计并合成1对特异性引物,应用PCR方法对河南省周边牛场576头荷斯坦牛进行检测,并对所有个体扩增出的DNA片段测序。[结果]PCR检测的576头奶牛中有2头FⅪ携带者和1头FⅪ缺陷症患者,与DNA序列测定结果一致,等位基因突变率为0.3%,携带率为0.3%,患病率为0.2%。[结论]建立了荷斯坦牛FⅪ的PCR检测方法,该方法具有简便、快速、特异性好等优点,适合荷斯坦牛的大规模检测。
[Objective] This study established a method to detect Factor Ⅺ by polymerase chain reaction analysis.[Method]A pair of primers was designed and synthesized according to sequences of FⅪ gene in Holstein calves,published in Genbank. Polymerase chain reaction was used to analyze FⅪ deficiency of 576 Holstein calves in Henan,and the result was verified by DNA sequencing. [Result] We detect 576 cows,which include two carriers and one F Ⅺ deficiency,and the result was consistent with the DNA sequencing. The frequency of the FⅪ mutant allele was 0.3%,the carrier was 0.3%,the prevalence was 0.2%.[Conclusion]A method detecting FⅪ by polymerase chain reaction analysis was established. This method is not only simple and convenient,but also has a high accuracy and low cost,which is more suitable for large-scale FⅪ investigation.
