DEVELOPMENT OF INDIRECT AVIAN INFLUENZA ELISA TEST USING MALAYSIA LOCAL ISOLATE H5N2

ONG G.H.1, SURIANI M.N.1, SYAMSIAH A.S.1, KHAIRUL A.M.1, RAMLAN M.1, CHANDRAWATHANI P.1 AND MAIZAN M.2

ONG G.H.1, SURIANI M.N.1, SYAMSIAH A.S.1, KHAIRUL A.M.1, RAMLAN M.1, CHANDRAWATHANI P.1 AND MAIZAN M.2

Serological test such as Hemagglutination Inhibition (HI) is used to detect Avian Influenza (AI) subtype specific antibodies. Agar gel precipitin test (AGPT) and Enzyme-Linked Immunosorbent Assay (ELISA) are used to detect AI group specific antibodies. These tests are used for AI poultry monitoring purposes. Indirect (AI) ELISA has been successfully developed in Veterinary Research Institute (VRI), Ipoh, Malaysia. Two types of antigens were prepared for this development to coat them onto the microtiter wells of MaxiSorb flat bottom plate (Nunc). They are from infected allantoic fluid (AF) of chicken embryonated eggs and infected tissue culture fluid (TCF) of Madin Darby Cannie Kidney cells using local isolate of H5N2 designated A/duck/Malaysia/8443/04. Conjugate of anti-chicken Horseradish Peroxidase and chromogen of o-Phenylenediamine (oPD) and Substrate 30% Hydrogen Peroxide were used. Results were read using ELISA reader of OD490 with reference OD630.