title: Development of a SYBR Green quantitative polymerase chain reaction assay for rapid detection and quantification of infectious laryngotracheitis virus.
abstract: Infectious laryngotracheitis is an acute viral respiratory disease of chickens with a worldwide distribution. Sensitive detection of the causative herpesvirus is particularly important because it can persist in the host at a very low copy number and be transmitted to other birds. Quantification of viral genome copy number is also useful for clinical investigations and experimental studies. In the study presented here, a quantitative polymerase chain reaction (qPCR) assay was developed using SYBR Green chemistry and the viral gene UL15a to detect and quantify infectious laryngotracheitis virus (ILTV) in ILTV-inoculated chicken embryos or naturally infected birds. The specificity of the assay was confirmed using a panel of viral and bacterial pathogens of poultry. The sensitivity of the assay was compared with two conventional PCR assays, virus titration and an antigen-detecting enzyme-linked immunosorbent assay. The qPCR developed in this study was highly sensitive and specific, and has potential for quantification of ILTV in tissues from naturally and experimentally infected birds and embryos.
authors.affiliation: Asia-Pacific Centre for Animal Health, School of Veterinary Science, The University of Melbourne, Victoria, 3010, Australia.
authors.forename: Alireza
authors.initials: A
authors.lastname: Mahmoudian
authors.forename: Naomi C
authors.initials: NC
authors.lastname: Kirkpatrick
authors.forename: Mauricio
authors.initials: M
authors.lastname: Coppo
authors.forename: Sang-Won
authors.initials: SW
authors.lastname: Lee
authors.forename: Joanne M
authors.initials: JM
authors.lastname: Devlin
authors.forename: Philip F
authors.initials: PF
authors.lastname: Markham
authors.forename: Glenn F
authors.initials: GF
authors.lastname: Browning
authors.forename: Amir H
authors.initials: AH
authors.lastname: Noormohammadi
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002645:Chickens; D004279:DNA, Viral; D003959:Diamines; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D001352:Herpesvirus 1, Gallid / Q000302:isolation & purification*; D009930:Organic Chemicals; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology*; D011804:Quinolines; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D052160:Benzothiazoles; D004279:DNA, Viral; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I