Submitted to: American Association of Avian Pathologists
Publication Type: Proceedings
Publication Acceptance Date: 3/5/2014
Publication Date: 7/21/2014
Citation: Spatz, S.J., Garcia, M., Yu, Q., Zsak, L. 2014. Development of a high throughput TaqMan assay for the detection of infectious laryngotracheitis virus in vector vaccinated chickens. In: Proceedings of the American Association of Avian Pathologists meeting, July 25-29, 2014, Denver, Colorado. p. 885.
Technical Abstract: Infectious laryngotracheitis virus (ILTV) causes an acute, highly contagious upper-respiratory disease of chickens. Sensitive detection of the causative alphaherpesvirus is important in clinical investigations and experimental studies. In particular, it is essential to quantify the viral genome copy number of the challenge virus in vaccine studies. A reduction in viral load is a critical parameter in defining the efficacy of an ILTV vaccine. The object of this study was to develop and evaluate a real time quantitative polymerase chain reaction (qPCR) for the detection of infectious laryngotracheitis virus in tracheal and eye swabs from vectored and modified live vaccinated chickens. Purified deoxyribonucleic acid (DNA) and sets of oligonucleotides specific for the glycoprotein C gene of ILTV and the chicken collagen gene along with FAM- and VIC-labeled probes were used in a uniplex 96 well format. DNA was also purified in a 96 well format from suspended tracheal and eye swabs using magnetic bead technology. The sensitivity and reproducibility of the ILTV real-time PCR assay revealed its suitability for detection and quantitation of ILTV in the samples from both experimentally and clinically ILTV infected chickens.