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Establishment and Application of Liquid Chip Method for Rapid Detection of Seven Porcine (Sus scrofa) Reproductive Diseases |
WU Xu-Long1, XIAO Lu2,*, YANG Miao3, XIE Li3, AN Wei3, GUO Yang3, ZHANG Qin3, WANG Yin4, LIN Hua3,* |
1 Animal Epidemic Disease Detection Center, Chengdu Agricultural College, Chengdu 611130, China; 2 Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Sichuan Animal Science Academy, Chengdu 610066, China; 3 Key Laboratory of Food Safety Testing of Sichuan Province, Technology Center of Chengdu Customs, Chengdu 610041, China; 4 College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China |
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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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Received: 11 June 2022
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Corresponding Authors:
*yaanxiaolu@163.com; cdcclh@126.com
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