鼠源细胞TaqMan实时荧光定量PCR检测方法的建立

Establishment of a TaqMan Real-time PCR Assay for Detection of Murine Derived Cells

关键词:  鼠源细胞  荧光定量 PCR  线粒体16S rRNA  检测。

Key words:  murine cells  fluorogenic quantitative PCR  mitochondrial 16S rRNA  detection.

摘要:

为对鼠源细胞进行鉴别检测，以鼠线粒体16S rRNA 基因序列为靶位点设计特异性引物及探针，建立实时荧光定量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，针对鼠源细胞基因组荧光定量PCR扩增曲线良好，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45.3拷贝。本试验建立的荧光定量PCR检测方法能够有效地对鼠源细胞进行快速检测，为细胞质量控制提供了有效方法。

Abstract:

To establish a fluorogenic quantitative PCR method, which could identify the cells of murine origin, specific primers along with probe were designed with the mitochondrial 16S rRNA gene sequence of murine, and the specificity alongwith sensitivity of the method were evaluated. The established detection method could specially detect murine 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 45.3 copies. The established fluorescence quantitative PCR detection method can effectively detect murine derived cells rapidly, providing an effective method for cell quality control.