title: Development and comparison of a Primer-Probe Energy Transfer based assay and a 5' conjugated Minor Groove Binder assay for sensitive real-time PCR detection of infectious laryngotracheitis virus.
abstract: In this study the design and development of two real-time PCR assays for the rapid, sensitive and specific detection of infectious laryngotracheitis virus (ILTV) DNA is described. A Primer-Probe Energy Transfer (PriProET) assay and 5' conjugated Minor Groove Binder (MGB) method are compared and contrasted. Both have been designed to target the thymidine kinase gene of the ILTV genome. Both PriProET and MGB assays are capable of detecting 20 copies of a DNA standard per reaction and are linear from 2×10(8) to 2×10(2)copies/μl. Neither PriProET, nor MGB reacted with heterologous herpesviruses, indicating a high specificity of the two methods as novel tools for virus detection and identification. This study demonstrates the suitability of PriProET and 5' conjugated MGB probes as real-time PCR chemistries for the diagnosis of respiratory diseases caused by ILTV.
authors.affiliation: School of Biological Sciences, Queen's University Belfast, Belfast BT9 7BL, Northern Ireland, United Kingdom. m.mcmenamy@qub.ac.uk
authors.forename: M J
authors.initials: MJ
authors.lastname: McMenamy
authors.forename: J
authors.initials: J
authors.lastname: McKillen
authors.forename: B
authors.initials: B
authors.lastname: Hjertner
authors.forename: I
authors.initials: I
authors.lastname: Kiss
authors.forename: A
authors.initials: A
authors.lastname: Yacoub
authors.forename: M
authors.initials: M
authors.lastname: Leijon
authors.forename: C
authors.initials: C
authors.lastname: Duffy
authors.forename: S
authors.initials: S
authors.lastname: Belák
authors.forename: M
authors.initials: M
authors.lastname: Welsh
authors.forename: G
authors.initials: G
authors.lastname: Allan
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D001352:Herpesvirus 1, Gallid / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004279:DNA, Viral