Diagnostic PCR for detection of hyperparasitism in cotton mealybug. PCR was performed with P. unfasciativentris-specific PCR primers, Hyp-ITS1F1 and ITS1-5.8sR1 to amplify a 755 bp ITS1 fragment. A, M, DNA marker, lane 1, DNA from identified Promuscidea unfasciativentris, lane 2, Aenasius bambawalei, lane 3, Phenacoccus solenopsis (3), lane 4, negative control; B, lanes 1-10, DNA from P. solenopsis adults collected from cotton fields and used in the diagnostic PCR. PCR bands (755 bp) in lanes 1, 3, 4, and 8 indicate the mealybugs were hyperparasitized by P. unfasciativentris while the empty lanes (2, 5, 6, 7, 9 and 10) indicate the mealybugs were free from the hyperparasitoid.