Evaluation Of The Recombinant Vaccinia Virus Immunogeniccity For MERS-CoV S And Preliminary Explored The Serun Antibody Detection Method

In 2012, a novel coronavirus (CoV) named middle east respiratory syndrome coronavirus, isolated from a man who presented with acute pneumonia in SaudiArabia. The common presenting symptoms of MERS include fever, cough,dyspnea,myalgia, chest pain,other clinical symptoms,infection complication with ARDS and acute ren-al failure. Up to 1 June 2015, Globally a total number of 1153 laboratory confir-med cases have been reported to the WHO, including 431 fatalities,mortality rate as high as 37%.MERS-CoV is the novel coronavirus infected with human lower respiratory tract after SARS-CoV, severely threated human health.MERS-CoV is an enveloped positive-sense single-stranded RNA virus MERS-CoV, belongs to lineage C of the genus Betacoronavirus (CoV) in the family Cor-onaviridae.MERS-CoV encoded structural protein include Spike protein,Nucleocaps-id protein,Membrane protein and Envelpoe protein. The spike protein is a specificity glycoprotein of MERS-CoV, no cross-reaction with other coronaviruses,assembled as trimers that constitute the typical crown-like peplomers on the surface of MERS-CoV;The spike protein is a povital role in host cell receptor recognization and virus entry host cell,and include important neutralize epitope that can induce robust neutra-lize antibody.Zoonotic transmission and human to human transmission indicated that it’s necessary to establish a specificity,sensitivity and convenient diagnostic method for MERS-CoV,and develop a vaccine prevent human from MER-SCoV infection.To solve above problems,the vaccina virus Tiantan strain was selected as virus vector to construct the recombination vaccinia virus expressing S1、SQ of MERS-CoV,evaluated the immunogenicity of the recombination virus. The recombinat virus was selected to express S1 antigen in primary CEF cells,establish indirect ELISA of MERS-CoV sera antibody dection based on S1 antigen,provide experimental data for diagnostic reagent and vaccine.The major results are summarized as follow:First, we constructed the recombination vaccinia virus expressing S1、SQ of MERS-CoV, identificated the recombination virus by PCR, indirect IFA and western blot.the results indicated that S1,SQ were accurately inserted into the vaccinia virus vector.Second, evaluated the immunogenicity the recombination virus expressing MERS-CoV S1,SQ. The result indicated that recombinat virus induced humoral immune response and cellular immune response in mice.Third, abundantly expressed S1 antigen, preliminary established an optimal condition of expression MERS-CoV diagnostic antigen and an indirect ELISA of MERS-CoV sera dection method. The results show that the purify of S1 antigen above as high as 90%, has preferable specificity.In conclusion,we expressed and prepared MERS-CoV S1 antigen, established an indirect ELISA method of MERS-CoV sera antibody detection; In future MERS-CoV may be occur in our country, The results will provide technical support and be the basis of the diagnose.Evaluation the immunogenicity of RVVMERS-S1, RVVM-ERS-SQ show that recombination virus can induced immune response, provided important data for the choice of target antigen in the future.