Monoclonal antibodies (MCA) produced against infectious
laryngotracheitis virus (ILTV) of chickens reacted in
western blotting experiments with several different ILTV
protein bands in the absence of tunicamycin which inhibits
carbohydrate synthesis. Most of the MCA lost their
reactivity in western blotting experiments when extracts of
tunicamycin-treated ILTV CELC were used, suggesting their
specificity for carbohydrate-based epitopes. In an indirect
immunofluorescence test most of the MCA bound primarily to
cytoplasmic antigens except some MCA which bound primarily
to nuclear antigens. Additivity ELISA was also performed to
study whether MCA are against the same epitope or different
epitopes.
The polymerase chain reaction (PCR) was developed as a
diagnostic technique for detection of ILTV using primers
made from a portion of the ILTV thymidine kinase gene. The
647-basepair amplified ILTV PCR product was labeled to
create a non-radioactive, biotinylated DNA probe.
Hybridization was performed using the probe to detect ILTV.
Both PCR and hybridization detected ILTV, and neither
hybridization nor PCR gave positive results with any other
pathogen. Hybridization was specific for ILTV, However,
slight hybridization occurred with CELC DNA when relatively
relaxed conditions were used.
In another experiment, diagnostic tests to detect ILTV
in tracheas of experimentally-infected chickens, including
the indirect fluorescent antibody test (IFAT),
immunoperoxidase (IP), virus isolation (VI), histopathology,
PCR, and hybridization, were performed and compared. Using
virus isolation as a reference, the sensitivity and
specificity of the tests were calculated. The IP test and
IFAT performed better than any other test used in this
study. / Graduation date: 1995
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/34948 |
| Date | 22 November 1994 |
| Creators | Abbas, Ferhat, 1962- |
| Contributors | Andreason, James R. Jr |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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