DEVELOPMENT OF MONOCLONAL FLUORESCENT ANTIBODY AGAINST COXSACKIE A16 VIRUS AND ITS APPLICATION IN CELL PLATFORM FOR VIRUS ISOLATION AND IDENTIFICATION

Jihong Rao*, Shuhua Zhang*, Kang Wang*, Mengjuan Yang*, Shumin Ge*, #, Liying Zhang**

*School of Life Science and Technology, Changchun University of Science and Technology - **College of Animal Technology Science, Jilin University, 130062, China

Objective: To establish a direct fluorescent antibody detection assay for use as an auxiliary experimental method in the isolation and identification of Coxsackievirus A16 (CA16). 

Methods: We purified 4E3 by chromatographic column. The antibody was combined with fluorescein isothiocyanate (FITC) by stirring. This titre and purified antibodies were identified by indirect ELISA and SDS-PAGE, respectively. The most suitable working concentration and specificity of the labelled fluorescent antibody was determined by direct immunofluorescence and gradient dilution methods, respectively. 

Result: SDS-PAGE indicted that purified antibodies had been harvested and that the titre of antibody reached 1:12800. The assay results shown that the optimal working concentration of the direct FITC antibody was diluted to 1:32. This direct FITC antibody showed no cross-reactivity with other common viruses, such as Eenterovirus 71(EV71) or Coxsackie virus B3(CB3), indicating that this antibody was specific.

Conclusion: The results indicate that the direct immunofluorescent antibody can be used for the rapid detection of CA16 on the cell platform. This method can provide relevant intuitive detection data, and is thus of great significance for virus isolation and identification.