Simultaneous detection of five notifiable viral diseases of cattle by single-tube multiplex real-time RT-PCR

Journal of Virological Methods
Kerstin WernikeMartin Beer

Abstract

Multiplexed real-time PCR (qPCR) assays enable the detection of several target genes in a single reaction, which is applicable for simultaneous testing for the most important viral diseases in samples obtained from ruminants with unspecific clinical symptoms. Here, reverse transcription qPCR (RT-qPCR) systems for the detection of bovine viral diarrhoea virus (BVDV) and bluetongue virus (BTV) were combined with an internal control system based on the beta-actin gene. Additionally, a background screening for three further major pathogens of cloven-hoofed animals reportable to the World Organisation for Animal Health, namely foot-and-mouth disease virus, epizootic haemorrhagic disease virus, and Rift Valley fever virus, was integrated using the identical fluorophore for the respective RT-qPCR assays. Every pathogen-specific assay had an analytical sensitivity of at least 100 genome copies per reaction within the multiplex approach, and a series of reference samples and clinical specimens obtained from cattle, but also from small ruminants, were detected reliably. The qPCR systems integrated in the background screening were even not influenced by the simultaneous amplification of very high BVDV and BTV genome copy numbers. The newl...Continue Reading

References

Aug 15, 1997·Nucleic Acids Research·J BrownieS Little
Apr 3, 1999·Systematic and Applied Microbiology·S T TayL R Krumholz
Mar 9, 2002·Nucleic Acids Research·Ian M MackayAndreas Nitsche
Mar 11, 2004·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·I M Mackay
Sep 17, 2005·Animal Health Research Reviews·Volker MoennigAnn Lindberg
Feb 8, 2006·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·R N GunsonW F Carman
Jan 2, 2007·Journal of Virological Methods·J F ToussaintK De Clercq
Dec 21, 2007·Journal of Virology·Donna M TscherneCharles M Rice
Mar 29, 2008·Schweizer Archiv für Tierheilkunde·M HofmannB Thür
Nov 4, 2008·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·R N GunsonW F Carman
Mar 17, 2009·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·William C WilsonJames O Mecham
Jul 14, 2010·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Yong-Il ChoKyoungjin J Yoon
Sep 9, 2010·Expert Review of Vaccines·Stéphan ZientaraGiovanni Savini
Dec 4, 2010·Journal of Virological Methods·Paul Francis Horwood, Timothy John Mahony
Mar 5, 2011·Preventive Veterinary Medicine·Patrick PresiDagmar Heim
May 17, 2011·Preventive Veterinary Medicine·N James Maclachlan
Jun 15, 2011·Research in Veterinary Science·G SaviniM Domingo
Jun 15, 2011·Viruses·Tetsuro Ikegami, Shinji Makino
Aug 26, 2011·Australian Veterinary Journal·Ae KessellPa Windsor
Sep 13, 2011·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Bernd HoffmannMartin Beer
Oct 19, 2011·Veterinary Microbiology·Corinne SailleauStéphan Zientara
Mar 2, 2012·Emerging Infectious Diseases·Bernd HoffmannMartin Beer
Dec 25, 2012·Research in Veterinary Science·P G ToyeC A L Oura
Aug 21, 2013·Preventive Veterinary Medicine·T J D Knight-Jones, J Rushton

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Citations

Jun 21, 2016·Transboundary and Emerging Diseases·T J D Knight-JonesW Vosloo
Apr 27, 2018·Journal of Clinical Microbiology·Eman AnisRebecca P Wilkes
Apr 17, 2019·PLoS Neglected Tropical Diseases·Emmanuel H HikufeAdrianatus Maseke
Aug 31, 2019·Transboundary and Emerging Diseases·Velizar BumbarovOran Erster
Apr 20, 2018·MSphere·Ann M GuggisbergAudrey R Odom John
Jul 5, 2018·Transboundary and Emerging Diseases·Kore SchlottauBernd Hoffmann
Jan 23, 2021·Journal of Zoo and Wildlife Medicine : Official Publication of the American Association of Zoo Veterinarians·Jeremy P OrangeJason K Blackburn
Dec 2, 2021·Transboundary and Emerging Diseases·Jørgen Steen Agerholm, Kerstin Wernike

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