利用PCR-SSCP检测荷斯坦奶牛瓜氨酸血症

Detecting Citrullinemia in Holstein Calves using PCR-SSCP analysis

【目的】建立荷斯坦奶牛瓜氨酸血症的PCR-SSCP检测方法。【方法】根据GenBank公布的荷斯坦奶牛精氨酸琥珀酸合成酶基因(Ass)第5外显子核苷酸序列(登录号:M2619),设计并合成1对特异性引物,应用PCR-SSCP方法检测北京市周边牛场173头荷斯坦奶牛瓜氨酸血症的隐性基因,并利用PCR-RFLP和DNA序列测定验证检测结果的准确性。【结果】PCR-SSCP检测的173头奶牛中有1头是瓜氨酸血症携带者,与PCR-RFLP和DNA序列测定结果一致,携带率为0.58%。【结论】建立了荷斯坦奶牛瓜氨酸血症的PCR-SSCP检测方法,该方法简单快捷、准确性高、成本低廉,适合荷斯坦奶牛的大规模检测。

【Objective】 The study established a method to detect Citrullinemia by polymerase chain reaction-single strand conformation polymorphism analysis.【Method】 A pair of primers were designed and synthesized according to nucleotide sequences of the exon 5 for argininosuccinate synthesize gene in Holstein Calves,published in Genbank and under accession number：M2619.Polymerase chain reaction-single strand conformation polymorphism was used to analyze exon 5 for argininosuccinate synthesize of 173 Holstein Calves in Beijing, and the results were verified by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing. [Result] One of the 173 Holstein Calves was Citrullinemia carrier,and the carrying rate was 0.58%. This result was consistent with that of the polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing. [Conclusion] A method detecting Citrullinemia by polymerase chain reaction-single strand conformation polymorphism 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 Citrullinemia investigation.