Kajian teknik imunositokimia untuk deteksdi virus Avian Influenza pada preparat apus darah burung Puyuh (Coturnix japonica)

SOLFAINE, Rondius, Prof. drh. Hastari Wuryastuty, M.Sc., Ph.D

2008 | Tesis | S2 Sain Veteriner

Pada akhir tahun 2003 sampai sepanjang tahun 2004 telah terjadi wabah avian influenza di Indonesia dan jutaan ayam petelur/pedaging komersial, ayam lokal dan burung puyuh mengalami kematian. Beberapa metode diagnosa untuk deteksi antibodi (hemagglutination inhibition/enzyme-linked immunosorbent assay) dan deteksi antigen (isolasi virus pada telur ayam bertunas dan polymerase chain reaction) telah dikembangkan untuk diagnosa secara cepat dan akurat terhadap virus avian influenza. Tujuan dari penelitian ini adalah mengkaji metode imunositokimia streptavidin-biotin (SB) untuk memvisualisasi lokasi seluler virus avian influenza dalam preparat apus darah burung puyuh dengan antibodi monoklonal spesifik terhadap nukleoprotein virus influenza tipe A. Hasil penelitian menunjukkan bahwa metode deteksi imunositokimia SB secara konsisten dapat mendeteksi virus influenza tipe A dalam preparat apus darah burung puyuh yang secara klinis atau secara serologis diduga menderita avian influenza. Hasil pewarnaan positif terhadap nukleoprotein virus diindikasikan sebagai agregat merah kecoklatan. Berdasarkan hasil penelitian dapat disimpulkan bahwa metode imunositokimia streptavidin-biotin dapat diaplikasikan untuk mendeteksi virus avian influenza dalam preparat apus darah burung puyuh.

At the end of the year of 2003 through the year of 2004, fatal avian influenza (AI) outbreaks occurred in Indonesia and millions commercial layer/broiler, local poultry and quails had been killed during the outbreaks. Several diagnostic methods either using indirect detection (hemagglutination inhibition/enzyme-linked immunosorbent assay) and direct antigen detection (virus isolation and polymerase chain reaction) methods have been developed and used for early and accurate diagnosis of AI. The objective of this study was to utilize immunocytochemistry method of streptavidin-biotin (SB) in order to visualize cellular location of AIV in the quail blood smears using monoclonal antibody specific for AIV type A nucleoprotein. The results show that immunocytochemistry detection method of SB consistently detects AIV type A in blood smears of quails that clinically and serologically suffering from AI. The immunocytochemistry SB positif staining of nucleoprotein are indicated by reddish-brown discoloration. Based on the results it can be concluded that immunocytochemistry SB method can be applied for detecting AIV in blood smears.

Kata Kunci : Avian influenza,Preparat apus darah,Imunositokimia streptavidin biotin,SB,immunocytochemistry streptavidin-biotin (SB), blood smears, avian influenza