Comparative Sensitivity Analysis Between two methods for Species Differentiation and Interspecies Cross Contamination in Animal Cell culture

Background and Aims : The present article focuses on design and development of a highly sensitive and convenient approach for rapid detection of animal species and cross contaminations quickly during cell cultures as most important document for manufacturing working cell bank system. This test is one of the four most important documents during implementing the banking system. By using this modified test also one of the major risks in cell culture laboratories, cross- contamination and misidentifications of cell lines will be also under test. Materials and Methods : A PCR _RFLP assay was optimized based on the use of a pair of primers that anneal to a portion the cytochrome b gene in all the species. The amplification product was digested with a panel of six restriction enzyme and the pattern derived was resolved on 3% high resolution agarose gel for 2 species, human & primate. As a control test we used from conventional method iso enzyme assay. Results : This protocol produced a unique restriction pattern and the origin of this animal cells resulted to be confirmed by this analysis. Its sensitivity in detecting interspecies cross contamination was at least 100 pg DNA/reaction, which was sufficient for detection of each species of DNA. Conclusion : For the last few years the RAZI vaccine and serum research institute in IRAN has been involved in developing such an ideal Iranian vaccine based on local Iranian material. The method developed in this study will provide a useful tool for the authentication of animal species comparable and most time consuming, cost benefit to other conventional analysis. Using the method (PCR-RFLP) developed during this research not only significant differences between human and non human also cross- contamination between different cell lines can simply distinguish.