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Detecting wMel Wolbachia in field-collected Aedes aegypti mosquitoes using loop-mediated isothermal amplification (LAMP).

Gonçalves DDS., Hooker DJ., Dong Y., Baran N., Kyrylos P., Iturbe-Ormaetxe I., Simmons CP., O'Neill SL.

BackgroundThe World Mosquito Program uses Wolbachia pipientis for the biocontrol of arboviruses transmitted by Aedes aegypti mosquitoes. Diagnostic testing for Wolbachia in laboratory colonies and in field-caught mosquito populations has typically employed PCR. New, simpler methods to diagnose Wolbachia infection in mosquitoes are required for large-scale operational use.MethodsField-collected Ae. aegypti mosquitoes from North Queensland were tested using primers designed to detect the Wolbachia wsp gene, specific to the strain wMel. The results were analysed by colour change in the reaction mix. Furthermore, to confirm the efficiency of the LAMP assay, the results were compared to the gold-standard qPCR test.ResultsA novel loop-mediated isothermal amplification (LAMP) colorimetric test for the wMel strain of Wolbachia was designed, developed and validated for use in a high-throughput setting. Against the standard qPCR test, the analytical sensitivity, specificity and diagnostic metrics were: sensitivity (99.6%), specificity (92.2%), positive predictive value (97.08%) and negative predictive value (99.30%).ConclusionsWe describe an alternative, novel and high-throughput method for diagnosing wMel Wolbachia infections in mosquitoes. This assay should support Wolbachia surveillance in both laboratory and field populations of Ae. aegypti.

More information Original publication

DOI

10.1186/s13071-019-3666-6

Type

Journal article

Publication Date

2019-08-01T00:00:00+00:00

Volume

12

Addresses

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Keywords

Animals, Aedes, Wolbachia, DNA Primers, Colorimetry, Nucleic Acid Amplification Techniques, Genes, Bacterial, Queensland, Mosquito Vectors