Mosquito Feeding Assay for Malaria Transmission

Mosquitoes were fed on gametocytaemic blood from volunteers enrolled in a CHMI transmission study via either direct skin feeding or membrane feeding assays [18 (link)]. Mosquitoes were dissected for evaluation of oocysts on day 8 or 9 after feeding assay. Midguts were collected in 180 µL of DNA Tissue Lysis buffer (Roche Diagnostics, Australia) for PCR analysis, or were stained with 0.5% mercurochrome for visualization of oocysts by microscopy prior to collection in lysis buffer and PCR analysis.

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Wang C.Y., McCarthy J.S., Stone W.J., Bousema T., Collins K.A, & Bialasiewicz S. (2018). Assessing Plasmodium falciparum transmission in mosquito-feeding assays using quantitative PCR. Malaria Journal, 17, 249.

Publication 2018

DNA Tissue Lysis buffer

Assay Blood Buffer Chmi Diagnostics Membrane Mercurochrome Microscopy Mosquitoes Oocysts Skin Tissue Transmission Volunteers

Corresponding Organization :

Other organizations : University of Queensland, QIMR Berghofer Medical Research Institute, Radboud University Nijmegen, Radboud University Medical Center

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Variable analysis

independent variables

Feeding method (direct skin feeding or membrane feeding assays)

dependent variables

Oocyst count on mosquito midguts (evaluated on day 8 or 9 after feeding assay)

control variables

Gametocytaemic blood from volunteers enrolled in a CHMI transmission study
Mosquito dissection and collection of midguts in DNA Tissue Lysis buffer (Roche Diagnostics, Australia) for PCR analysis
Staining of midguts with 0.5% mercurochrome for visualization of oocysts by microscopy prior to collection in lysis buffer and PCR analysis

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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