The H9N2 subtype of low-pathogenic avian influenza viruses (LPAIV) is a widespread pathogen of poultry that can also infect humans. The characterization of viral infections is a complex process, involving clinical, pathological, and virological investigations. The aim of this study was to adapt and optimize an immunohistochemical (IHC) technique developed for LPAIVs specifically for the detection of H9N2 virus antigens in infected tissues. Twenty-one-day-old broiler chickens were inoculated with three different strains of H9N2 virus by different infection routes (i.e., intranasal-intratracheal and intravenous) or co-infected with infectious bronchitis virus (IBV) and observed for 11 days post infection. The suggested IHC protocol was modified: (i) DAB (diamino-benzidine) was substituted with AEC (3-amino-9-ethyl carbazole) as chromogen; and (ii) indirect two-step immune reactions of monoclonal primary and peroxidase-labeled anti-mouse secondary antibodies were used instead of avidin–biotin complexes. Avian influenza virus antigen appears as a red precipitate in the nuclei of affected cells but can also be identified in the cytoplasm. Mild hyperemia and congestion were observed in the trachea, air sac, and lungs of the challenged birds, and fibrinous exudate was found at the bifurcation in a few cases. Neither gross pathological nor IHC lesions were found in the control group. Using the optimized protocol and an associated scoring scheme, it was demonstrated that the H9N2 strains tested exhibited respiratory and urinary tract tropism irrespective of the route of inoculation. On day 5, viral antigen was detected in the respiratory tract and kidney in 30–50% of the samples. On day 11, no IHC signal was observed, indicating the lack of viral replication. Slight differences in viral antigen expression were found between the different H9N2 virus strains, but, in contrast to highly pathogenic avian influenza (HPAI), no viral antigen was detected in the brain and pancreas. Thus, IHC can be considered as an informative, visual addition to the toolkit for the characterization of H9N2 LPAIV infections.

A H9N2 altípusú influenzavírus növekvő elterjedtsége, gazdasági, járványtani, humánegészségügyi jelentősége miatt a szerzők irodalmi adatok alapján összefoglalják, rendszerezik az altípussal kapcsolatos aktuális ismereteket. Napjainkban a madárinfluenza-vírusok közül a H9N2 altípusúak a legelterjedtebbek a baromfifajokban. Bár korábban a vadmadárvonulások, valamint az élőállatpiacok voltak a fertőződés forrásai, az elmúlt évtizedekben a már baromfihoz adaptálódott vírusok egyre nagyobb földrajzi területen váltak endémiássá és egészítették ki a legjelentősebb baromfi légzőszervi kórokozók körét. A H9N2 altípusú vírusok kivétel nélkül alacsony pathogenitásúak, ugyanakkor egyéb kórokozókkal társfertőzésben, baromfiban már súlyos megbetegedéseket eredményeznek. A H9N2 vírusok zoonotikus szempontból kis kockázatot jelentenek, ugyanakkor számos más, már jelentős arányú elhalálozást okozó madárinfluenza-vírus belső génjeit a H9N2 vírus adta.