The spined soldier bug, Podisus maculiventris (Say), is an important biological control agent of agricultural and forest pests; and it preys on eggs and larvae of lepidopteran and coleopteran species. Genetic variability among field samples collected from Michigan, Mississippi, Missouri, Florida and established laboratory colonies was examined using Amplified Fragment Length Polymorphism-Polymerase Chain Reaction (AFLP-PCR). Four AFLP primer pairs generated a total of 340 molecular markers for evaluation. Results from Analysis of Molecular Variance showed that the majority of the genetic variation occurred within populations (individuals from each sample site). Nei's method indicated reduced genetic diversity in laboratory populations compared to field populations. No major differences or deficiencies were apparent among the field samples from different areas or among the laboratory reared samples. We conclude that field populations are panmictic and laboratory reared spined soldier bug could be useful as biological control agents in the field.