Study On The Characteristics Of Coxsackievirus A16Humoral Immunity Responses

Objetive:Coxsackie virus type16group A (CA16) and enterovirus71(EV71) are two major pathogenic agents of hand, foot and mouth disease (HFMD). The current study found that When HFMD outbreak, EV71and CA16often alternate or popular together. However, due to the CA16infection and repeatedly cross reactivity with other enteroviruses, makes the CA16complex, humoral immune response characteristics of CA16is unclear. This study to constrct CA16VP1full-length and the truncated antigen, using NEIBM ELISE method, study on the characteristics of human body against coxsackievirus A16humoral immune response.Method:Prokaryotic expression plasmid pET21b-CA16VP11-297aa, pET32a-CA16VP141-297aa, pPEPTIDE2-CA16VP11-60aa, pPEPTIDE2-CA16VP145-58aa and pET32a-CA16VP161-297aa were constructed, transformed into E.coli BL21(DE3) for expession under induction of IPTG, The expressed recombinant proteins were purified by nickel ion affinity chromatography, The expressed recombinant proteins CA16VP11-297was identified by ELISA,using NEIBM indirect ELISA method to detect anti CA16VP1antibodies with95serums of the elderly in taiyuan city(mean age:61.0±14.0years) and142serums of blood donation members in Shanghai city (average age:35.5±11.2years), and to analysis by competitive ELISA.Result:1.Successfully constructed five kinds of CA16VP1the complete and truncated recombinant plasmid and expressed in escherichia coli, obtained the five kinds of VP1and truncated recombinant proteins, including pPEPTIDE2-CA16VP11-60, pPEPTIDE2-CA16VP145-58exists in the form of soluble protein, pET32a-CA16VP161-297,pET32a-CA16VP141-297,pET21b-CA16VP11-297exists in the form of inclusion body.2. NEIBM indirect ELISA test results showed that: positive rate of reaction of taiyuan elderly serums with CA16VP11-297, CA16VP141-297, CA16VP11-60, CA16VP145-58and CA16VPl61-297antigens were76.84%,40.00%,33.68%,31.57%and30.53%; Shanghai blood donation member crowd serums anti-CA16VP11-297positive rate was59.15%..3. Competitive ELISA results of Anti-CA16VP11-297strong reaction serums of46taiyuan older showed that:CA16VP11-297, CA16VP141-297, the CA16VP11to60, CA16VP145-58and CA16VP161-297antigen inhibition rate median were73.50%,53.75%,21.00%, 20.50%and15.50%. Competitive ELISA results of Anti-CA16VP11-297strong reaction serums of84Shanghai blood donation members showed that:CA16VP11-297, CA16VP141-297, the CA16VP11-60, CA16VP145-58and CA16VP161-297antigen inhibition rate median were86.00%、86.00%、82.50%、62.00%and28.50%.4. The competition patterns and composition rario of taiyuan elderly serums and Shanghai blood donation members serums were obviously different.Conclusion:1.The elderly crowd in Taiyuan city and the adult population in Shanghai city have higher CA16and/or CA16related antibodies.2. Testing inhibition situation of CA16VP1the full-length and the truncated antigen with anti-CA16VP1of taiyuan elderly and Shanghai crowd are obviously different, suggest the two types of people infected with different CA16and/or CA16related viruses.3. Human anti-CA16antibody reaction mainly focusing on cross epitope based antigen.