Absolute quantification of Bovine Viral Diarrhea Virus (BVDV) RNA by the digital PCR technique

7th Brazilian Congress on Metrology

Flatschart, R.B.^1 ; Almeida, D.O.^1 ; Heinemann, M.B.^2 ; Medeiros, M.N.^1 ; Granjeiro, J.M.^1 ; Folgueras-Flatschart, A.V.^1

INMETRO-Instituto Nacional de Metrologia, Qualidade e Tecnologia, Duque de Caxias-RJ, Brazil^1

EV/UFMG-Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte-MG, Brazil^2

关键词: Absolute quantification;    Biopharmaceuticals;    Bovine viral diarrhea virus;    Calibration standard;    Detection and enumerations;    Digital pcr;    Real-time PCR;    Reference material;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/575/1/012038/pdf
DOI  :  10.1088/1742-6596/575/1/012038

来源: IOP

【 摘 要 】

The quality control of cell lines used in research and industry is critical to ensure confidence in experimental results and to guarantee the safety of biopharmaceuticals to consumers. The BVDV is a common adventitious agent in many cell lines. We preliminarly evaluate the use of Digital Droplet PCR (ddPCR) for the detection and enumeration of genome copies of BVDV in cell culture and on FBS. The application of a commercial Real-Time PCR kit with the ddPCR technique was successful on different matrices. The technique allowed the absolute quantification of the genome without the use of calibration standards, suggesting its promising application on the development of reference materials for quantification of nucleic acids.