Advance Publication by J-STAGE Japanese Journal of Infectious Diseases A novel reporter gene assay for pyrogen detection

This study explored the feasibility of a novel reporter gene assay (RGA) for pyrogen detection using RAW264.7 cells stably transfected with the NF-κB reporter gene as a pyrogenic marker. The RGA could detect different types of pyrogens, including the lipopolysaccharide of gram-negative bacteria, the lipoteichoic acid of gram-positive bacteria, and the zymosan of fungi, and a good dose-effect relationship was observed in terms of NF-κB activity. The limits of detection of the RGA to those pyrogens were 0.03 EU/ml, 0.001 μg/ml, and 1μg/ml, respectively. The method had good precision and accuracy and could be applied to many products bevacizumab, etanercept, group A and group C meningococcal conjugate vaccine, diphtheria, tetanus, pertussis (acellular, component), poliomyelitis (inactivated) vaccine, and imject alum adjuvant]. The results of this study suggest that the novel RGA has a wide pyrogen detection spectrum and is sufficiently sensitive, stable, and accurate for various applications.