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.