MGB双探针实时荧光PCR检测牛白细胞粘附缺陷病方法的建立

MGB Two Probe Assay for Detecting Bovine Leukocyte Adhesion Deficiency in Cattle by Real-Time PCR

从福建某奶牛场250头进口荷斯坦牛采血提取牛血液DNA,根据已知牛染色体上CD18编码基因序列383位碱基由A变为G而引起牛白细胞粘附缺陷病设计特异性野生型和突变型探针,建立实时荧光定量PCR检测方法用于牛白细胞粘附缺陷病的基因分型检测。结果检出2头母牛为牛白细胞粘附缺陷病携带者,检出率为0.8%,没有发现患病牛。结果证明本研究建立的实时荧光定量PCR检测方法是一种敏感性和特异性很高的筛选奶牛牛白细胞粘附缺陷病有害基因的新方法。

A MGB two probe assay based on real-time PCR was developed and applied to detect bovine leukocyte adhesion deficiency in Holstein cows. A total of 250 Holstein cows imported from foreign countries were genotyped. Two cows were identified as BLAD carriers, corresponding to the carrier frequency 0.8%, there was no patient cow. The results showed that MGB two probe assay based on real-time PCR is a new sensitive and specific assay for genotyping BLAD.