Pathogen detection
Samples collected from patients and wild mice were detected for common rodent and tick-borne zoonotic pathogens. PCR was used to assay bacterial pathogens with total nucleic acid such as Rickettsia, Coxiella, anaplasmosa and Borrelia burgdorferi. Hantavirus, Seoul virus, novel Bunyavirus, Semliki Forest virus, Dabieshan orthohantavirus, new Alongshan virus (tick-borne flavivirus found for the first time in Liaoning Province) and other viral pathogens were detected by PCR with retrotranscriptional products. PCR reaction system: 20ul, reaction procedure: 94℃ 5min, 94℃ 30s, 54℃ 20s, 72℃ 30s, 72℃ 10min. The sequence of primers mentioned above is detailed in Appendix 1. The amplified PCR products were subjected to agarose gel electrophoresis, and the positive products were sent to Sangong Bioengineering (Shanghai) Co., Ltd. for sequencing.