Laboratory Protocol for Assessing Aedes Mosquito Exposure and Arboviral Infection v1

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This protocol describes laboratory methods for assessing human exposure to Aedes aegypti mosquito bites and detecting arboviral RNA in human blood samples collected in Northern Tanzania. The protocol combines two complementary laboratory approaches: an enzyme-linked immunosorbent assay (ELISA) for measuring human IgG antibodies against the Nterm-34 kDa salivary peptide of Aedes aegypti and a one-step multiplex real-time reverse transcription polymerase chain reaction (RT-qPCR) assay for detecting dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV). The ELISA uses dried blood spot (DBS) samples to quantify anti-Nterm-34 kDa IgG responses based on the difference in optical density between antigen-coated and no-antigen wells, providing an indirect measure of human exposure to Aedes mosquito bites. The RT-qPCR method involves viral RNA extraction from whole blood samples and detection of arboviral RNA using a commercially available manufacturer's kit according to the manufacturer's instructions. Together, these laboratory methods support the assessment of mosquito exposure and arboviral infection in the study population. The protocol describes the sample preparation, ELISA procedure, RNA extraction, RT-qPCR assay setup, quality control procedures, and interpretation of laboratory results.

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