Optimization and validation of multiplex PCR method for cell species identification
10.13200/j.cnki.cjb.004655
- VernacularTitle:细胞种属鉴别多重PCR法的优化及验证
- Author:
Xueling WU
1
Author Information
1. Cell Collection and Research Center, National Institutes for Food and Drug Control, Beijing 102629, China
- Publication Type:Journal Article
- Keywords:
Cell species;
Identification;
Multiplex PCR method;
Mitochondria
- From:
Chinese Journal of Biologicals
2026;39(02):180-188
- CountryChina
- Language:Chinese
-
Abstract:
Objective To further optimize and validate the original cell species identification multiplex PCR method to make it applicable to different testing agencies and laboratories.Methods The primer sequences of pigs, Chinese hamsters, African green monkeys and rats, the methods of nucleic acid extraction, the ratios of primers and reference cell mixed genomic DNA, the PCR procedures of amplification and the DNA template dosage were optimized. The sensitivity and cross contamination limits of the optimized method were validated, and reviewed and verified across different laboratories.Results The optimized method was successfully used for cell identification and detection of 10 species, with a sensitivity of 50-5 000 cells and a detection level of cross contamination of 1∶100-1∶10 000. Four different laboratories could use this method to successfully detect cells.Conclusion The optimized cell species identification multiplex PCR method has good specificity and ability to detect cell cross contamination, and can be used for cell species identification in different laboratories with good robustness, which will provide effective technical means for quality control of cells used in production and testing.
