Assessment of Aedes albopictus reference genes for quantitative PCR at different stages of development

Members of the Aedes genus of mosquitoes are widely recognized as vectors of viral diseases. Ae.albopictus is its most invasive species, and are known to carry viruses such as Dengue, Chikugunya and Zika. Its emerging importance puts Ae.albopictus on the forefront of genetic interaction and evolutio...

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Bibliographic Details
Published in:PloS one 2018-03, Vol.13 (3), p.e0194664-e0194664
Main Authors: Dzaki, Najat, Azzam, Ghows
Format: Article
Language:English
Subjects:
Aedes aegypti
Aedes albopictus
Algorithms
Aquatic insects
Biological evolution
Biology and Life Sciences
Blood
Cell culture
Cell cycle
Dengue
Dengue fever
Developmental stages
Embryos
Evolutionary genetics
Females
Gene expression
Genes
ILK protein
Insects
Instars
Introduced species
Invasive species
Lepidoptera
Macaca mulatta
Males
Medicine and Health Sciences
Molecular biology
Mosquitoes
Normalizing
Research and Analysis Methods
Stability analysis
Vector-borne diseases
Vectors
Viral diseases
Viruses
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Summary:Members of the Aedes genus of mosquitoes are widely recognized as vectors of viral diseases. Ae.albopictus is its most invasive species, and are known to carry viruses such as Dengue, Chikugunya and Zika. Its emerging importance puts Ae.albopictus on the forefront of genetic interaction and evolution studies. However, a panel of suitable reference genes specific for this insect is as of now undescribed. Nine reference genes, namely ACT, eEF1-γ, eIF2α, PP2A, RPL32, RPS17, PGK1, ILK and STK were evaluated. Expression patterns of the candidate reference genes were observed in a total of seventeen sample types, separated by stage of development and age. Gene stability was inferred from obtained quantification data through three widely cited evaluation algorithms i.e. BestKeeper, geNorm, and NormFinder. No single gene showed a satisfactory degree of stability throughout all developmental stages. Therefore, we propose combinations of PGK and ILK for early embryos; RPL32 and RPS17 for late embryos, all four larval instars, and pupae samples; eEF1-γ with STK for adult males; eEF1-γ with RPS17 for non-blood fed females; and eEF1-γ with eIF2α for both blood-fed females and cell culture. The results from this study should be able to provide a more informed selection of normalizing genes during qPCR in Ae.albopictus.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0194664