Diagnosis of newcastle disease virus infections and monitoring of immune responses using enzyme immunoassays
1990
Burgess, G.W. (James Cook Univ., of North Queensland (Australia). Graduate School of Tropical Veterinary Science and Agriculture) | Lamichhane, C.M. | Malcolm, K.M.
Detection of Newcastle disease viral antigens can assist with the identification of new isolates. In addition the specific reactions of the antigens with selected monoclonal antibodies can be used to accurately type the strains of virus. Antigen detection studies in this laboratory to date have concentrated on the use of immunoperoxidase techniques. Antigens have been detected in suspension using ELISA. They have also been detected in cell cultures, allantoic cells, embryonic tissues and in tissues from adult birds using substrates which are precipitated in the infected cells. An antigen detection ELISA uses globulin derived from hyperimmune rabbit serum to capture the antigen to the plastic plate and monoclonal antibodies followed by peroxidase conjugated anti-mouse globulin as the indicator system. This assay is capable of detecing Indonesian strains of NDV directly from homogenates of the organs of experimentally infected chickens with a 64-128 fold increase in sensitivity when compared to the haemagglutination assay. The assay in its present form may be performed in two and a half hours using precoated plates, with the results being read by eye. A technique for staining of NDV antigens in cell cultures has been used extensively to screen monoclonal antibodies in this laboratory. The assay uses a bovine kidney cell culture (MDBK) infected with NDV. When lentogenic strains are grown, the media must contain a proteolytic enzyme. Indirect immunoperoxidase has been used to study the distribution of NDV antigen in the tissues of avian embryos. Avian embryos were infected both with Australian lentogenic strains of virus and panels of viruses in both Indonesia and Nepal. The embryos were fixed overnight in formalin and tissues as well as allantoic membrane were embedded in wax.
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