Direct nucleic acid analysis of mosquitoes for high fidelity species identification and detection of Wolbachia using a cellphone

PLoS Neglected Tropical Diseases
Sanchita BhadraAndrew D Ellington

Abstract

Manipulation of natural mosquito populations using the endosymbiotic bacteria Wolbachia is being investigated as a novel strategy to reduce the burden of mosquito-borne viruses. To evaluate the efficacy of these interventions, it will be critical to determine Wolbachia infection frequencies in Aedes aegypti mosquito populations. However, current diagnostic tools are not well-suited to fit this need. Morphological methods cannot identify Wolbachia, immunoassays often suffer from low sensitivity and poor throughput, while PCR and spectroscopy require complex instruments and technical expertise, which restrict their use to centralized laboratories. To address this unmet need, we have used loop-mediated isothermal amplification (LAMP) and oligonucleotide strand displacement (OSD) probes to create a one-pot sample-to-answer nucleic acid diagnostic platform for vector and symbiont surveillance. LAMP-OSD assays can directly amplify target nucleic acids from macerated mosquitoes without requiring nucleic acid purification and yield specific single endpoint yes/no fluorescence signals that are observable to eye or by cellphone camera. We demonstrate cellphone-imaged LAMP-OSD tests for two targets, the Aedes aegypti cytochrome oxidase I ...Continue Reading

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Citations

Jun 5, 2019·Ecology and Evolution·Aditi KulkarniJiannong Xu
Jan 11, 2021·BMC Public Health·Maria Angelica CarrilloSilvia Runge-Ranzinger
May 21, 2021·MSphere·Sanchita BhadraAndrew D Ellington
Mar 14, 2019·Chemical Reviews·Siyuan S Wang, Andrew D Ellington

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Methods Mentioned

BETA
nucleic acid amplification
PCR
fluorescence imaging
nucleic
nucleic acid purification
pregnancy test

Software Mentioned

LightCycler
OSD
Primer Explorer
MUSCLE BLAST
LAMP
- OSD
NUPACK
MUSCLE MUltiple Sequence Comparison by

Related Concepts