Aphids, which feed solely on plant phloem sap, have developed symbiotic associations with bacteria that provide them with the amino acids that are lacking in phloem. Three soybean aphid (Aphis glycines Mat samura) populations were screened for the presence of Buchnera aphidicola and three common species of secondary aphid symbionts (Serratia symbiotica, Hamiltonella defensa, and Regiella insecticola). Diagnostic polymerase chain reaction and subsequent DNA sequencing showed the presence of two species of symbiotic bacteria present in all three soybean aphid populations tested: B. aphidicola and Arsenophonus sp. Although Buchnera is commonly found in aphids, Arsenophonus is most commonly found in whiteflies (Hemiptera: Aleyrodidae), making the soybean aphid unique among aphids that have been tested for the presence of Arsenophonus.