Abstract
In this study, we examined various brain suspension concentrations and viral loads in Neuro-2a cell cultures using 20 rabies-positive bovine samples. The reproducibility of results varied: 65% showed consistent outcomes across all concentrations, while 35% disagreed in at least one. Viral titers ranged from less than 25 × 101 to 25 × 103.50 TCID50/mL, with 20% below 25 × 101 TCID50/mL. Concentrations between 5% and 20% yielded over 90% agreement in positive results, but at 30%, agreement dropped from 85% to 50%. Cell confluence was successfully maintained at 5%, 10%, and 20%, while concentrations of 30% and above led to confluence loss. Low viral loads also negatively impacted reproducibility. These results suggest that sample concentration has a direct influence on preservation of cell confluence and that low viral loads may influence the reproducibility of the rabies tissue culture infection test (RTCIT).
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Acknowledgments
We thank the Instituto Pasteur, São Paulo, Brazil, for financial support (IP 04/2014). The data presented in this manuscript were described in a monograph work by Tatiane de Cássia Pardo de Souza.
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Funding was provided by Instituto Pasteur, São Paulo, Brazil support (IP 04/2014).
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This study was approved by the Institutional Ethics Committee for Animal Use (CEUA-IP, protocol number 05/2014).
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de Souza, T.d.C.P., de Sousa e Silva, G., de Oliveira Fahl, W. et al. Evaluation of the effect of sample suspension concentration and viral load on the outcome of the rabies tissue culture infection test. Arch Virol 169, 3 (2024). https://doi.org/10.1007/s00705-023-05922-3
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DOI: https://doi.org/10.1007/s00705-023-05922-3