Methodology

Determination of suitable housekeeping genes for normalisation of quantitative real time PCR analysis of cells infected with human immunodeficiency virus and herpes viruses

Sarah Watson¹ , Sarah Mercier¹ , Chris Bye² , John Wilkinson³ , Anthony L Cunningham¹ and Andrew N Harman¹

¹  Centre for Virus Research, Westmead Millennium Institute, Sydney, Australia

²  The Howard Florey Institute, University of Melbourne, Melbourne, Australia

³  Biotron Limited, Centre for Immunology, St. Vincent's Hospital, Sydney, Australia

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Virology Journal 2007, 4:130doi:10.1186/1743-422X-4-130

Published:	3 December 2007

Abstract

The choice of an appropriate housekeeping gene for normalisation purposes has now become an essential requirement when designing QPCR experiments. This is of particular importance when using QPCR to measure viral and cellular gene transcription levels in the context of viral infections as viruses can significantly interfere with host cell pathways, the components of which traditional housekeeping genes often encode. In this study we have determined the reliability of 10 housekeeping genes in context of four heavily studied viral infections; human immunodeficiency virus type 1, herpes simplex virus type 1, cytomegalovirus and varicella zoster virus infections using a variety of cell types and virus strains. This provides researchers of these viruses with a shortlist of potential housekeeping genes to use as normalisers for QPCR experiments.