title: Reliable reference gene selection for quantitative real time PCR in Haemonchus contortus.
abstract: 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.
authors.affiliation: Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Lecová
authors.affiliation: Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Růžičková
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G61 1QH, Scotland, UK.
authors.forename: Roz
authors.initials: R
authors.lastname: Laing
authors.affiliation: Max Planck Institute for Chemical Ecology, Department of Entomology, Jena D-07745, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Vogel
authors.affiliation: Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Szotáková
authors.affiliation: Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Lukáš
authors.initials: L
authors.lastname: Prchal
authors.affiliation: Department of Pharmacology and Toxicology, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Lamka
authors.affiliation: Department of Pharmacology and Toxicology, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Ivan
authors.initials: I
authors.lastname: Vokřál
authors.affiliation: Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Skálová
authors.affiliation: Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy, Hradec Králové, Czech Republic. Electronic address: matousp7@faf.cuni.cz.
authors.forename: Petra
authors.initials: P
authors.lastname: Matoušková
mesh_terms: D000818:Animals; D000871:Anthelmintics / Q000494:pharmacology; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D006189:Haemonchus / Q000187:drug effects / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards
chemical_list: D000871:Anthelmintics
keywords: Anthelmintic; Haemonchus contortus; Reference gene; qPCR