From qualitative to quantitative insect metabarcoding: an in tandem multilocus mosquito identification methodology

Kassela, K.; Kouvela, A.; De Courcy Williams, M.; Konstantinidis, K.; Rosa-Freitas, M. G.; Nearchou, A.; Gatzidou, E.; Veletza, S.; Boulougouris, G. C.; Dovrolis, N.; Karakasiliotis, I.

doi:10.1101/2020.11.22.393140

bioRxiv · 10.1101/2020.11.22.393140

From qualitative to quantitative insect metabarcoding: an in tandem multilocus mosquito identification methodology

Kassela, K.·Kouvela, A.·De Courcy Williams, M.·Konstantinidis, K.·Rosa-Freitas, M. G.·Nearchou, A.·Gatzidou, E.·Veletza, S.·Boulougouris, G. C.·Dovrolis, N.·Karakasiliotis, I.

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Abstract

In the era of emergence and re-emergence of vector-borne diseases, a high throughput trap-based insect monitoring is essential for the identification of invasive species, study of mosquito populations and risk assessment of disease outbreaks. Insect DNA metabarcoding technology has emerged as a highly promising methodology for unbiased and large-scale surveillance. Despite significant attempts to introduce DNA metabarcoding in mosquito or other insect surveillance qualitative and quantitative metabarcoding remains a challenge. In the present study, we have developed a methodology of in-tandem identification and quantification using cytochrome oxidase subunit I (COI) combined with a secondary multilocus identification and quantification involving three loci of 28S ribosomal DNA. The presented methodology was able to identify individual species in pools of mosquitoes with 95.94% accuracy and resolve with high accuracy (p = 1,{chi} 2 = 2.55) mosquito population composition providing a technology capable of revolutionizing mosquito surveillance through metabarcoding. The methodology, given the respective dataset, has the potential to be applied to various small animal populations.

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Kassela, K., Kouvela, A., De Courcy Williams, M., Konstantinidis, K., Rosa-Freitas, M. G., Nearchou, A., Gatzidou, E., Veletza, S., Boulougouris, G. C., Dovrolis, N., Karakasiliotis, I.. 2020-11-22. From qualitative to quantitative insect metabarcoding: an in tandem multilocus mosquito identification methodology. https://doi.org/10.1101/2020.11.22.393140

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