Establishment of ELISA method with recombinant Hantaviral nucleocapsid protein as antigen and its application for Hantavirus serologic typing
|School||Fourth Military Medical University|
|Keywords||Hantavirus Recombinant nucleocapsid protein Serotype DOB type Enzyme - linked immunosorbent assay|
Hemorrhagic fever with renal syndrome caused by hantavirus worldwide popular natural foci of disease . The number of cases of the disease each year in China accounted for more than 90% of the total number of cases in the world , and also an upward trend in recent years , this study aims to build a fast , accurate and easy to spread Genotyping , clinical diagnosis and epidemic areas detected . Hantavirus nucleocapsid protein amino-terminal 1 to 117 of the B cell recognition of a plurality of antigenic determinant , and can induce a high level of the humoral immune . We use this feature, the basis of the Elgh study , the prokaryotic vector expressing strains of E.coli BL-21 (DE3) changed BL21-CodonPlus, increase identification arginine , leucine and isoleucine tRNA genes , improve the yield of expression , to overcome the disadvantages of the expression yield unstable . The principle application polyhistidine chain and nickel ion chelate column containing nickel ion with a poly-histidine chain NP antigen affinity purified , has been expressed in E.coli , with the person fourth Military Medical University, a master's degree thesis disease five boron Han ketone poison u order variegate , Seoul , Dobra , Puumala and Xinnuo Bo ), nucleocapsid protein , and the recombinant The protein as attending to write original , to establish indirect EUSA law . Of the 8 were male rib Qing , 3 negative serum E : USA law and IFA law gingival comparison coincidence rate hundred. 9%. China 's Shaanxi , Shandong , Hebei and Beijing of 44i5 City 's 241 ' serum of patients with hemorrhagic fever with renal syndrome detection results of hantavirus infection mainly Hantaan and Seoul ; 4O copies polymorphism sexual type ; 4g typing antigen DOBjlgisili , reaction , suggesting that there serotypes DOB serological evidence of in the country has not yet seen such a . In short with other hantavirus detection methods ( such as immunofluorescence and plaque reduction neutralization test ) compared to using recombinant nucleocapsid protein lazy original the EUSA law faint kinds of security, economic and easy . Fast, easy way to promote the application at the grassroots level laboratory . The emergence of multi- type serotype , further research is needed to find a reasonable explanation . Meanwhile, the discovery of the new serotypes of hemorrhagic fever with renal syndrome vaccine research and disease surveillance tight corner d will have far-reaching implications .