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Method Validation of Quantitative Real-time RTPCR for Newcastle Disease Virus (NDV) and Comparison of the Method with Other Quantitative Methods

초록

영어

Newcastle disease virus (NDV) has come into the spotlight because of its oncolytic property for cancer therapy and usefulness as a viral vector for vaccine development (1, 2). For the clinical use, the NDV production is performed in cell-based system rather than traditional egg-based system for compliance with GMP/GLP (4). In this study, the rapid detection and quantitation method for the NDV produced in cell-based system was developed. The SYBR Green I-based real-time RT-PCR was designed with conventional cheap RT-PCR kit by targeting F gene of the NDV LaSota strain. The developed method was validated by the contents of specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantitation (LOQ) and robustnesss (4-7). The validation results are satisfied with the predetermined acceptance criteria. The validated method was applied to quantitate the virus samples produced in cell-based production system. And the method was compared with other quantitating methods (3, 7). The comparison results showed that the real-time RT-PCR method would be well adequate to detect and quantitate the whole virus particles containing both infective and uninfective rapidly.

저자정보

  • Juno JANG Dept. of Biotechnology, Korea University, Seoul, 136-701.
  • Sung-Hwan HONG Dept. of Biotechnology, Korea University, Seoul, 136-701.
  • Ik-Hwan KIM Dept. of Biotechnology, Korea University, Seoul, 136-701.

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