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You J.,Shanghai Biosci Biotechnology Co. | Huang L.,Shanghai Biosci Biotechnology Co. | Zhuang J.,Shanghai Biosci Biotechnology Co. | Mou Z.,Shanghai Biosci Biotechnology Co.
Food Science and Biotechnology | Year: 2014

A multiplex real-time PCR method for discriminating deer and common domestic species, including cattle, goat, horse, donkey, pig, and chicken was developed. Species-specific primer pairs were designed and used to produce different size DNA fragments with diverse melting temperature (T m) values. The specificity and sensitivity of these primer pairs were separately confirmed using simplex real-time PCR analysis. Multiplex real-time PCR analysis was performed using combined primers, yielding distinct melting curve profiles for each species. The sensitivity limit of the multiplex PCR method was evaluated. Trace DNA of other species in deer DNA could be identified. Common deer products, including blood, meat, and antler were tested using this multiplex PCR method and different species of deer and domestic animals were identified. The rapid multiplex real-time PCR assay described herein is a specific, sensitive, and reliable method for high-throughput authentication of deer and domestic animal products. © 2014 The Korean Society of Food Science and Technology and Springer Science+Business Media Dordrecht. Source


Zhou C.,Shanghai Biosci Biotechnology Co. | Gao X.,Shanghai Biosci Biotechnology Co. | He S.,Shanghai Biosci Biotechnology Co. | Zhuang J.,Shanghai Biosci Biotechnology Co. | And 2 more authors.
Hematology | Year: 2016

Objectives: Monoclonal anti-human blood group A (51A8) and B (63B6) antibody reagents were prepared using the serum-free technique. The aims of this research were to characterize the serum-free reagents and prove their reliabilities in routine use. Methods: Experiments including antigen–antibody agglutination testing, stability testing, SDS–PAGE, protein and IgM quantification, flow cytometry, and variable domain sequencing were performed to characterize the anti-A (51A8) and anti-B (63B6) reagents. Over 12 000 samples were tested using these reagents as routine blood grouping reagents. Results: Serum-free anti-A (51A8) and anti-B (63B6) reagents were stable in longitudinal and accelerated testing, and their high purity was shown in SDS–PAGE and IgM quantification. These reagents have high specificity to red blood cells in serologic agglutination testing and flow cytometric analysis. A1 and A2 subgroup antigens can be distinguished clearly by patterns of flow cytometric histograms. No discrepancy was found in clinical trials of 12 000 samples. Discussion: To reduce the risk of being affected by any animal additives, a serum-free culture system was applied to get mass-production of monoclonal anti-A/B antibodies. The high specificity and the high purity of the reagents were verified by the lab experiments. Conclusion: Lab research and clinical trial showed that serum-free monoclonal anti-A (51A8) and anti-B (63B6) reagents meet the requirements of routine blood grouping reagents. Moreover, these reagents featured ultra-high purity that is missing in other commercial counterparts, and therefore are recommended as more environment-friendly reagents. © 2016 Informa UK Limited, trading as Taylor & Francis Group Source

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