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目的:建立一种优于传统方法的、简单快速和灵敏度高的细胞种属鉴别及种属间细胞交叉污染的PCR检测方法。方法:以细胞色素b(cytochrome b)作为研究内容,设计并优化针对10种不同动物种属的引物,进行单种属引物和混合种属引物PCR反应的比较研究。结果:本文设计出可通过混合引物单PCR方式高效、敏感地检测10种常见动物种属细胞,并有效判断种属间细胞交叉污染的PCR检测方法。结论:本方法明显优于传统的细胞种属鉴定及种属间细胞交叉污染的方法,能显著提高对细胞资源库细胞、生产用细胞基质及治疗性细胞产品的质控能力。
OBJECTIVE: To establish a PCR assay that is superior to the traditional methods to identify cell species and cross-species cell-to-cell cross-contamination easily and quickly. Methods: Using cytochrome b as the research content, we designed and optimized the primers for 10 species of different animals, and compared the PCR reactions of single species primers and mixed species primers. Results: In this study, we designed a PCR assay that can effectively and sensitively detect 10 kinds of common animal species cells by hybrid primer single-PCR and effectively determine the cross-contamination of cells between species. Conclusion: This method is obviously superior to the traditional methods of identification of cell species and cross-contamination of interspecies cells, which can significantly improve the quality control ability of cell resource bank, production cell matrix and therapeutic cell products.