Abstract：With the development and progress of the pig industry in China, higher expection are required for the differential diagnosis and big data analysis of diseases. Here, a novel method was established, which promoted the development and application of high-throughput detection technology in animal pathogen detection. In detail, specific primers and probes were designed for the Pseudorabies virus (PRV)、Japanese encephalitis virus (JEV)、Classical swine fever virus (CSFV)、Porcine circovirus type 2 (PCV-2)、Porcine reproductive and respiratory syndrome virus (PRRSV)、Porcine parvovirus (PPV)、African classical swine fever virus (ASFV) 7 pathogens of swine (Sus scrofa) reproductive diseases. After optimizing the reaction conditions, coupling the probe with microspheres, and hybridization capture reaction, a liquid chip detection method to detect 7 pig reproductive diseases at the same time was successfully established and applied to clinical practice. The results showed that the specific target gene could be captured and detected in the liquid environment, the detection limit of 7 specific pathogens was 103 copies/μL. The method had good specificity and no cross reaction with other pathogens. The established method was used to detect the 65 samples collected from large-scale pig farms in Sichuan province. A total of 49 positive samples were detected, with a positive rate of 75.38% (49/65). The positive rate of mixed infection of 2 or more pathogens was 23.08% (15/65). The results of single PCR detection were consistent with the established method, which confirmed that the established liquid chip detection method can be applied to clinical detection. This study provides a powerful technical reserve for rapid differential diagnosis of multiple pathogens of swine disease, and provides a reference for the further development of high-throughput detection technology.
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