Based on high-throughput qPCR (HT-qPCR) quantitative detection technology, it was used to quantify 96 virulence factor genes (VFGs) of four typical zoonotic pathogenic bacteria (K. pneumoniae, A. baumannii, E. coli and S. enterica), including 120 pairs of primers, and participated in the encoding of five major virulence factor types (Toxin, Adherence, Secretion system, Immune evasion/invasion、Iron uptake）。 VFG Chip has the advantages of high throughput, low sample size, low cost, short cycle time, and simple operation, and can be widely applied to various types of samples, providing a powerful tool for the comprehensive quantification of VFGs in the environment, thus providing a new idea for assessing the health risk of zoonotic pathogens in the environment.