Development Of Quantitative Methods For Detection Of The Nucleic Acid And Protein Of Equine Arteritis Virus

Equine viral arteritis (EVA) was a disease that caused by equine arteritis virus (EAV). To develop a method for EAV detection, antigen capture enzyme-linked immunosorbent assay (AC-ELISA) was developed on the base of the monoclonal antibodies 2B3 and 2B9 on protein level, and the results showed that the detection limit of the AC-ELISA method for the EAV Bucyrus strain was 3.86 TCID50, and no cross reaction to regular viruses of equids with high specificity. The triplicate tests of the AC-ELISA method were showed good stability. Meanwhile, the real-time RT-PCR assay based on Eva Green was developed on nucleotide level to be used to detect EAV with a pair of specific primers for ORF7 by EAV Bucyrus strain. The results showed that the Ct value and viral load of EAV had linearity from 102 copies/μL to 107 copies/μL by the developed assay. The detection limit of the RT-PCR assay was 101.5 TCID50/mL for virus and 10 copies/μL for the standard plasmid. The CVs of intra-and inter-assay for the assay were all lower than 2% with good reproducibility.In all, the two methods were developed base for the research of EAV on etiology.