Rapid and simultaneous detection of antibodies against three feline viruses using colloidal gold immunochromatographic assay with smartphone integration
A multiplex colloidal gold immunochromatographic assay (mCGIA) was developed for the simultaneous detection of feline calicivirus (FCV), feline panleukopenia virus (FPV), and feline herpesvirus (FHV) antibodies in feline serum. The expressed and purified recombinant FCV-VP1, FPV-VP2, and FHV-gD proteins exhibited strong antigenic reactivity. The mCGIA test strip achieved rapid (< 15 min) and specific detection, showing no cross-reactivity with other feline viral antibodies. The assay achieved a low visual limit of detection (vLOD) of 1:1 × 10 5 for FCV and 1:1 × 10 4 for FPV and FHV antibodies. Stability testing revealed 90% retention of detection capability after 4 weeks of storage at 37 °C. Reproducibility was robust, with intra- and inter-assay coefficients of variation (CVs) of less than 7% and 13%, respectively. The smartphone-based portable reader enabled quantitative detection with wide linear ranges (from 1:1000 to 1:100000 for FCV, 1:250 to 1:10000 for FPV and FHV) and excellent linearity (FCV: R 2 = 0.992, FPV and FHV: R 2 = 0.991). Compared with indirect ELISA using 164 clinical serum samples, the assay showed 98.17% (FCV), 98.78% (FPV), and 97.56% (FHV) concordance rates, respectively. This novel mCGIA platform provides a rapid, cost-effective tool for sero-surveillance, vaccine evaluation, and assisting clinical diagnosis, enhancing feline health management. Graphical
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