ABSTRACT
<p><b>OBJECTIVE</b>To establish a quantitative assay for enterovirus 71, this can be used in detecting the virus content during vaccine development and production.</p><p><b>METHODS</b>We established the method of quantitative assay for EV71 by using double antibody sandwich ELISA. The sensitivity, accuracy,precision and specificity of the method were evaluated.</p><p><b>RESULTS</b>We developed an ELISA method to quantitative assay for EV71. The quantitation limit of the method is 0.23 microg/ml and the quantitation scope of the method is 7.32-0.23 microg/ml, the coefficient correlation is R2 = 0.9976; The method showed good accuracy, precision and specificity. The recovery is between 90%-110% and the variation coefficient is lower than 10%.</p><p><b>CONCLUSION</b>An ELISA method was developed for the quantitative assay of EV71 virus, which can be used for the rapid quantitative determination of EV71 virus during vaccine development and production.</p>
Subject(s)
Animals , Humans , Antigens, Viral , Allergy and Immunology , Cell Line , Enterovirus , Allergy and Immunology , Enterovirus Infections , Diagnosis , Allergy and Immunology , Virology , Enzyme-Linked Immunosorbent Assay , Methods , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovirus 71.</p><p><b>METHODS</b>Two peptides, SP55 and SP70, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to establish the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test.</p><p><b>RESULTS</b>Two high titered anti-VP1 antibodies secreted by the hybridoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1:16 respectively.</p><p><b>CONCLUSION</b>Two high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.</p>