Dissertation > Medicine, health > Clinical > Diagnostics > Laboratory diagnosis > Immunology Examination

Establishment of ELISA method with recombinant Hantaviral nucleocapsid protein as antigen and its application for Hantavirus serologic typing

Author LiuJingMei
Tutor XuDeZhong
School Fourth Military Medical University
Course Epidemiology
Keywords Hantavirus Recombinant nucleocapsid protein Serotype DOB type Enzyme - linked immunosorbent assay
CLC R446.6
Type Master's thesis
Year 2001
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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 .

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