The aim of this work was to identify reliable reference genes for expression studies in adult Haemonchus contortus. Eleven candidate genes were identified and the stability of their expression was assessed in adult males and females of two genetically divergent H. contortus isolates: drug-susceptible (ISE) and multi-drug-resistant (WR). Five genes with the most stable expression patterns were further assessed for suitability as reference genes in anthelmintic-treated H. contortus adults versus non-treated controls. We identified important differences in the expression of a number of candidate genes in anthelmintic-treated samples, confirming the need for careful validation of control genes for such experiments. We propose the use of multiple reference genes for expression studies in this species and found gpd, ama and far most suitable for adult H. contortus.

Department of Biochemical Sciences Charles University Prague Faculty of Pharmacy Hradec Králové Czech Republic

Department of Pharmacology and Toxicology Charles University Prague Faculty of Pharmacy Hradec Králové Czech Republic

Institute of Biodiversity Animal Health and Comparative Medicine College of Medical Veterinary and Life Sciences University of Glasgow Glasgow G61 1QH Scotland UK

Max Planck Institute for Chemical Ecology Department of Entomology Jena D 07745 Germany

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$a Reliable reference gene selection for quantitative real time PCR in Haemonchus contortus / $c L. Lecová, M. Růžičková, R. Laing, H. Vogel, B. Szotáková, L. Prchal, J. Lamka, I. Vokřál, L. Skálová, P. Matoušková,

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$a The aim of this work was to identify reliable reference genes for expression studies in adult Haemonchus contortus. Eleven candidate genes were identified and the stability of their expression was assessed in adult males and females of two genetically divergent H. contortus isolates: drug-susceptible (ISE) and multi-drug-resistant (WR). Five genes with the most stable expression patterns were further assessed for suitability as reference genes in anthelmintic-treated H. contortus adults versus non-treated controls. We identified important differences in the expression of a number of candidate genes in anthelmintic-treated samples, confirming the need for careful validation of control genes for such experiments. We propose the use of multiple reference genes for expression studies in this species and found gpd, ama and far most suitable for adult H. contortus.

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$a Růžičková, Michaela $u Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.

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$a Laing, Roz $u Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G61 1QH, Scotland, UK.

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$a Vogel, Heiko $u Max Planck Institute for Chemical Ecology, Department of Entomology, Jena D-07745, Germany.

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$a Szotáková, Barbora $u Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.

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$a Prchal, Lukáš $u Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.

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$a Lamka, Jiří $u Department of Pharmacology and Toxicology, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.

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$a Vokřál, Ivan $u Department of Pharmacology and Toxicology, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.

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$a Skálová, Lenka $u Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.

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$a Matoušková, Petra $u Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic. Electronic address: matousp7@faf.cuni.cz.

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