Selection of reference genes for quantitative PCR analysis in poultry redmite (Dermanyssus gallinae)

摘要

Poultry red mites (PRMs, Dermanyssus gallinae) are harmful ectoparasitesthat affect farmed chickens and cause serious economic losses in the poultry industryworldwide. Acaricides are used for PRM control; however, some PRMs have developedacaricide-resistant properties, which have indicated the need for different approaches forPRM control. Therefore, it is necessary to elucidate the biological status of PRMs todevelop alternative PRM control strategies. Quantitative polymerase chain reaction (qPCR)allows analysis of the biological status at the transcript level. However, reference genesare preferable for accurate comparison of expression level changes given the largevariation in the quality of the PRM samples collected in each farm. This study aimed toidentify candidate reference genes with stable expression levels in the different bloodfeeding states and life stages of PRMs. First, we selected candidates based on thefollowing criteria: sufficient expression intensity and no significant expressiondifference between fed and starved states. We selected and characterized seven candidatereference genes. Among them, we evaluated the gene expression stability between thestarved and fed states using RefFinder; moreover, we compared their expression levels ineach life-stage and identified two reference genes, Elongation factor1-alpha (ELF1A)-like and apolipophorins-like.Finally, we evaluated the utility of the candidates as reference genes, and the use ofELF1A-like and apolipophorins-like successfullynormalized ATP synthase subunit g -like gene expression. Thus,ELF1A-like and apolipophorins-like could be suitablereference genes in PRMs.