Protocol for arbovirus infection of Aedes aegypti by intrathoracic microinjection or artificial blood feeding

Here, we present a protocol for infecting adult female Aedes aegypti with arboviruses using intrathoracic microinjection or artificial blood feeding as alternative approaches. We describe steps for rearing mosquitos, virus preparation, intrathoracic microinjection, and artificial blood feeding. We then detail procedures for saliva collection, tissue dissection (midgut and carcass), and assessment of vector competence. The protocol provides practical guidance, critical experimental considerations, and validation steps to improve reproducibility and consistency across laboratory settings.