Ji Jingjing, Li Jiahao, Wu Minghui, Wang Bin, Lu Yu-Nan, Zhang Chuhan, Shan Yanke, Liu Fei
Sanya Institute of Nanjing Agricultural University, Joint International Research Laboratory of Animal Health and Food Safety of Ministry of Education, Single Molecule Biochemistry & Biomedicine Laboratory (Sinmolab), Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China.
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China.
Mikrochim Acta. 2025 Jul 23;192(8):510. doi: 10.1007/s00604-025-07373-w.
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 for FCV and 1:1 × 10 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 = 0.992, FPV and FHV: R = 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.
开发了一种多重胶体金免疫层析检测法（mCGIA），用于同时检测猫血清中的猫杯状病毒（FCV）、猫泛白细胞减少症病毒（FPV）和猫疱疹病毒（FHV）抗体。表达并纯化的重组FCV-VP1、FPV-VP2和FHV-gD蛋白表现出很强的抗原反应性。mCGIA试纸条实现了快速（<15分钟）和特异性检测，与其他猫病毒抗体无交叉反应。该检测法对FCV抗体的低视觉检测限（vLOD）为1:1×10，对FPV和FHV抗体为1:1×10。稳定性测试显示，在37℃储存4周后，检测能力保留90%。重复性良好，批内和批间变异系数（CVs）分别小于7%和13%。基于智能手机的便携式读数器能够进行定量检测，线性范围宽（FCV为1:1000至1:100000，FPV和FHV为1:250至1:10000）且线性良好（FCV：R = 0.992，FPV和FHV：R = 0.991）。与使用164份临床血清样本的间接ELISA相比，该检测法的符合率分别为98.17%（FCV）、98.78%（FPV）和97.56%（FHV）。这种新型的mCGIA平台为血清学监测、疫苗评估和辅助临床诊断提供了一种快速、经济高效的工具，增强了猫的健康管理。
