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Related Experiment Videos

Effective mosquito and arbovirus surveillance using metabarcoding.

J Batovska1, S E Lynch1, N O I Cogan1,2

  • 1Agriculture Victoria, AgriBio, Centre for AgriBioscience, Bundoora, Vic, Australia.

Molecular Ecology Resources
|April 19, 2017
PubMed
Summary

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Metabarcoding, a DNA sequencing technique, effectively identifies mosquito species and Ross River virus (RRV) in pooled samples. This method shows promise for large-scale vector surveillance and disease screening.

Area of Science:

  • Entomology
  • Molecular Biology
  • Epidemiology

Background:

  • Effective vector and arbovirus surveillance necessitates scalable and accurate screening methods.
  • Next-generation sequencing (NGS) offers high-throughput potential for modernizing vector surveillance.
  • Metabarcoding combines NGS with DNA barcoding for simultaneous species and pathogen detection.

Purpose of the Study:

  • To establish and validate a metabarcoding protocol for characterizing mosquito pools.
  • To screen mosquito pools for the presence of Ross River virus (RRV).
  • To assess the sensitivity and limitations of the metabarcoding approach for surveillance.

Main Methods:

  • Mosquito pools (up to 100 individuals) were analyzed using DNA metabarcoding.
  • Mitochondrial cytochrome c oxidase subunit I (COI) and RRV E2 gene regions were amplified.
Keywords:
cytochrome c oxidase subunit ICulicidaeDNA barcodingbulk samplepooled samplesvirus

Related Experiment Videos

  • Illumina MiSeq sequencing and bioinformatic analysis against a reference database were performed.

    • Main Results:

    • Metabarcoding successfully identified all mosquito species and RRV in tested samples.
    • The protocol detected species and RRV from a single mosquito in a pool of 1000 individuals.
    • Factors like primers, PCR cycles, and sample size influence species quantification.

    Conclusions:

    • Metabarcoding provides a sensitive and scalable alternative for vector and arbovirus surveillance.
    • The protocol is effective for identifying multiple mosquito species and specific viruses simultaneously.
    • Recommendations are provided for optimizing metabarcoding in future surveillance studies.