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 x 10(8) to 2 x 10(2) copies/mu l. 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. (C) 2011 Elsevier B.V. All rights reserved.

Keywords

Infectious laryngotracheitis virus; Real-time PCR; Primer-Probe Energy Transfer; Minor Groove Binder probe

Published in

Journal of Virological Methods
2011, volume: 175, number: 2, pages: 149-155
Publisher: ELSEVIER SCIENCE BV

SLU Authors

• Hjertner, Bernt

  • Queen's University Belfast

• Belak, Sandor

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Animal and Dairy Science
Veterinary Science

Publication identifier

• DOI: https://doi.org/10.1016/j.jviromet.2011.04.020

Permanent link to this page (URI)

https://res.slu.se/id/publ/46515