摘要
为对猴源细胞进行鉴别检测,以猴线粒体16SrRNA基因序列为靶标设计特异性引物及探针,建立了TaqMan荧光定量PCR检测方法,并评价该方法的特异性及敏感性。结果显示,所建立的检测方法特异性好,针对猴源细胞基因组扩增曲线良好,针对其他物种来源细胞基因组及生物制品原辅材料未出现特异性扩增曲线;敏感性高,基因拷贝数检出限度为10拷贝/μL,对Vero、MA104和Marc-145细胞的检出限度均可达到10个细胞。本研究建立的TaqMan荧光定量PCR检测方法能够有效地对猴源细胞进行快速检测,为猴源细胞鉴别检测提供了技术支持。
To establish a taqman real-time PCR method,which could identify the cells of monkey origin,specific primers along with probe were designed with the mitochondrial 16S rRNA gene sequence of monkey,and the specificity alongwith sensitivity of the method were evaluated.The established detection method could specially detect monkey cells without detecting any other animal cells along with raw and secondary materials of biological products.The detection limit of 16S rRNA gene copy number is 10 copies/μL,and the detection limit of Vero,MA104 and Marc-145 is 10 cells each.The established real-time PCR detection method can effectively detect monkey derived cells rapidly,providing an effective method for cell quality control.
作者
苏佳
白洪旭
赵炜
秦义娴
毛娅卿
薛青红
陈晓春
吴华伟
SU Jia;BAI Hong-xu;ZHAO Wei;QIN Yi-xian;MAO Ya-qing;XUE Qing-hong;CHEN Xiao-chun;WU Hua-wei(China Institute of Veterinary Drug Control,Beijing 100081,China)
出处
《中国兽药杂志》
2025年第8期8-14,共7页
Chinese Journal of Veterinary Drug
基金
中国兽医药品监察所兽药行业公益性重点专项“兽用生物制品外源病毒污染及非法添加检测技术研究”(GY202406)。
作者简介
苏佳,兽医师,博士,从事兽用生物制品检验及相关研究工作;通讯作者:陈晓春,E-mail:chunxiao1981@126.com;通讯作者:吴华伟,E-mail:314174205@qq.com。
