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Molecular and immunogenic characterization of BHK-21 cell line adapted CVS-11 strain of rabies virus and future prospect in vaccination strategy

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Abstract

Development of a cost effective quality vaccine is a key issue in rabies control programme in developing countries. With this perspective, in the present study, challenge virus standard (CVS)-11 strain of rabies virus was adapted to grow in BHK-21 cells, characterized, compared with other viruses including global vaccine strains and field isolates from Indian subcontinent and China at molecular level. This cell adapted virus was evaluated for the production of cost effective veterinary vaccine. The maximum virus titre achieved was 107 fluorescent focus unit (FFU)/mL at 10th passage level. There was no nucleotide difference in the nucleoprotein (N) and glycoprotein (G) genes after adaptation in cell line. Phylogenetic analysis showed that adapted virus was grouped with global vaccine strains, closest being with other CVS strains but distinct from the Indian field isolates. Global vaccine strains including cell adapted CVS-11 virus have 83–87 % identity at nucleotide level of G gene with Indian field viruses. Growth kinetics of cell culture adapted virus showed that the optimum virus titer (around 107 FFU/mL) could be obtained at around 48 h post infection by co-cultivation method using 0.1 multiplicity of infection inoculums at 37 °C. These findings can be used for up scaling of vaccine production. The protective efficacy of test vaccine produced using 106.95 FFU/mL cell culture harvest showed 1.17 IU/mL relative potency by NIH test. Further, adapted virus was found to be suitable for use in rapid fluorescent focus inhibition test.

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Acknowledgments

Authors are thankful to Dr V. K. Chaturvedi and Dr A. K. Tiwari and Director, IVRI for providing the necessary facilities. This work was in part funded by ICAR-Indian Veterinary Research Institute under the Project Code IXX08803.

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Authors and Affiliations

  1. Division of Biological Products, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, India

    Arunkumar C. Patel, Santhamani Ramasamy & Rabindra Prasad Singh

  2. Division of Biological Standardization, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, India

    Vikramaditya Upmanyu

  3. Division of Veterinary Biotechnology, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, India

    Praveen Kumar Gupta

  4. Division of Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, India

    Rajendra Singh

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  1. Arunkumar C. Patel
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Correspondence to Rabindra Prasad Singh.

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Patel, A.C., Upmanyu, V., Ramasamy, S. et al. Molecular and immunogenic characterization of BHK-21 cell line adapted CVS-11 strain of rabies virus and future prospect in vaccination strategy. VirusDis. 26, 288–296 (2015). https://doi.org/10.1007/s13337-015-0285-5

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  • DOI: https://doi.org/10.1007/s13337-015-0285-5

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