New SNP Markers to Identify Culex Complex Species (Diptera: Culicidae) by High-Resolution DNA Melting Analysis

The members from Culex pipiens complex are the primary vector of various arboviruses including West Nile virus and Eastern equine encephalitis in the United States. Within Cx. pipiens complex, there are three biotypes that differ largely in habitat, bloodmeal preference, mating behavior, and overwin...

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Bibliographic Details
Published in:Journal of medical entomology 2018-10, Vol.55 (6), p.1446-1452
Main Authors: Kim, Sungshil, Lee, Rahmi, Sim, Cheolho
Format: Article
Language:English
Subjects:
a high-resolution melting analysis
Amplification
Biomarkers
Biotypes
Culex pipiens f. molestus
Culex pipiens f. pipiens
Culex quinquefasciatus
Culicidae
Deoxyribonucleic acid
Diptera
DNA
Eastern equine encephalitis
Encephalitis
Genes
Genetic markers
High resolution
Mating behavior
Melting
MOLECULAR BIOLOGY/GENOMICS
Nucleotides
Overwintering
Polymerase chain reaction
Single-nucleotide polymorphism
Vector-borne diseases
Viruses
West Nile virus
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Summary:The members from Culex pipiens complex are the primary vector of various arboviruses including West Nile virus and Eastern equine encephalitis in the United States. Within Cx. pipiens complex, there are three biotypes that differ largely in habitat, bloodmeal preference, mating behavior, and overwintering strategy. The three biotypes are Cx. pipiens form molestus (F.) (Diptera: Culicidae), Cx. pipiens f. pipiens (L.) (Diptera: Culicidae), and Cx. pipiens quinquefasciatus (Diptera: Culicidae) (also known as Cx. quinquefasciatus). It is clear that additional genetic markers, molecular markers based on single nucleotide polymorphisms (SNPs), were identified from the Cx. pipiens complex species. Genomic DNA was extracted from adult females of f. pipiens and f. molestus and was amplified by polymerase chain reactions. Thirty out of the 100 primer pairs showed amplification and were used for SNP identification. All thirty genes contained biotype-specific SNPs: 10 were located on the Cx. pipiens complex genetic map from previous genetic studies. We also tested a high-resolution DNA melting analysis as a biotype identification method by examining the SNPs in the two genes (CPIJ005487 and CPIJ002074). Our method provides a high confidence for biotype determination among the three Cx. pipiens complex mosquitoes. Data deposition: NCBI SNP Database (ssSNP accession nos.: ss947848635–ss947848878)
ISSN:0022-2585
1938-2928
DOI:10.1093/jme/tjy120