Differentiation of Anopheles gambiae and An. arabiensis (Diptera: Culicidae) by ELISA using immunoaffinity-purified antibodies to vitellogenin.

Yolk proteins (vitellogenin and vitellin) proved to be excellent marker molecules for separating Anopheles gambiae Giles and An. arabiensis Patton, two morphologically indistinguishable members of the An. gambiae species complex. A rabbit polyclonal antibody directed against An. gambiae yolk proteins was made species-specific by removing immunoglobulins that crossreacted with An. arabiensis by immunoaffinity chromatography. The resultant antibody was 400 times more sensitive to An. gambiae and was employed as the secondary antibody in a modified double antibody "sandwich" ELISA, which also used monoclonal antibodies to anopheline vitellogenin as the primary or coating antibody. This ELISA easily differentiated soluble yolk protein samples from An. gambiae and An. arabiensis. A field study with 628 females of An. gambiae complex collected in western Kenya demonstrated that the ELISA results were 98.4% in agreement with the standard cytotaxonomic method.