荷斯坦奶牛瓜氨酸血症PCR-DHPLC检测方法建立与应用  

The development and application of PCR-DHPLC for detecting Holstein bovine Citrullinemia

作　　者：辛学谦 郑小龙 张晓文 王群 姜帆 孙涛 梁成珠 岳志芹 XIN Xue-qian;ZHENG Xiao-long;ZHANG Xiao-wen;WANG Qun;JIANG Fan;SUN Tao;LIANG Cheng-zhu;YUE Zhi-qin(Technology Center of Qingdao Customs District,Qingdao 266002,China)

机构地区：[1]青岛海关技术中心

出　　处：《中国兽医杂志》2019年第5期77-79,I0006,共4页Chinese Journal of Veterinary Medicine

基　　金：“十三五”国家重点研发计划子课题(2016YFD050110401)

摘　　要：为建立一种快速高通量的荷斯坦奶牛瓜氨酸血症(CN)的分子检测方法,根据CN是由于精氨酸琥珀酸(ASS)合成酶第86位精氨酸突变为终止密码子的遗传学基础,设计了1对引物对3种不同基因型的基因材料PCR扩增,利用Navigator software软件分析确定检测温度、洗脱浓度及梯度等条件,建立检测方法。对55份进境荷斯坦奶牛血样进行检测,发现有1例为隐性基因携带者,将PCR产物送测序,结果与DHPLC检测结果一致。研究表明,该方法可用于荷斯坦奶牛脊椎畸形综合征的检测,是一种有效的新方法。Citrullinemia(CN)is caused by a transition of C→T at codon 86 within exon 5 in the gene coding for argininosuccinate synthetase enzyme(ASS).To develop a rapid and high-flux molecular detection method for CN,one pair of specific primers was designed according to the gene sequence of bovine ASS gene.The PCR fragments of three different genotypes were analyzed by DHPLC under the conditions predicted by the Navigator software.55 blood samples from imported Holstein bovine were detected by the established protocol.There was one CN carrier in 55 samples.The result of sequencing of the PCR product of the CNcarrier was right.This study provids a more reliable and useful method for detecting CN and also offers a theoretical basis for molecular diagnosis in Holstein bovine.

关 键 词：瓜氨酸血症 变性高效液相色谱 荷斯坦奶牛 

分 类 号：S851.34[农业科学—预防兽医学]