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Background: Timely entomological and insecticide resistance monitoring is a key to generating relevant data for vector management. We investigated the insecticide susceptibility status of Anopheles gambiae s.l. in eight rural farming communities in Southern Gombe, Nigeria. Methods: Overall, 3–5 days-old adult female Anopheles mosquitoes reared from field-collected immature stages between September and November, 2014 were exposed to the diagnostic doses of pyrethroids, organophosphate and carbamate insecticides using the Center for Disease Control Bottle bioassay. The observatory knockdown time from exposure to each insecticide was recorded up to two hours. The dead mosquitoes were then identified morphological­ly and by molecular assays. Results: Mortality results showed resistance in An. gambiae s.l. populations to bendiocarb (2.3–100%), deltamethrin (39–70%), pirimiphos-methyl (65–95%), dichloro-diphenyl-trichloroethane (0–38.1%), permethrin (0–46.3%) and lambda-cyhalothrin (42.5–86.4%). The few cases of full susceptibility were observed from lamdacyhalothrin exposed population of An. gambiae s.l. in Banbam and Pantami respectively. An. gambiae 177 (45%) was significantly higher (P< 0.05) than An. arabiensis 64 (16.3%), An. coluzzii 34 (8.7%) and An. gambiae/An. coluzzii hybrid 78 (19.8%). Conclusion: A strong evidence of widespread resistance in the major malaria vector species in Southern Gombe to all common classes of insecticides is a justification for the State Malaria Elimination Programme to consciously con­sider incorporating insecticide resistance management strategies into control programs in order to sustain the future of current control interventions