Several criteria in B-scan and colour-coded Doppler ultrasound have been shown to discriminate malignant transformation of cervical lymph nodes. All these parameters tend to be operator dependent. We describe the first use of computer-assisted analysis of contrast-enhanced ultrasound images evaluating cervical lymphadenopathy in differentiation between benign and malignant lesions. The results from sonography were compared with the histologic findings in 60 patients. Subjective sonomorphologic parameters were evaluated and compared with quantitative criteria obtained from computer analysis after intravenous contrast medium (SonoVue) application to estimate diagnostic impact. Our results confirm that malignant lymph nodes are round in shape (p < 0.001) and show peripheral perfusion (p = 0.0089). Dynamic parameters revealed that malignant nodes were characterized by contrast medium wash-in time (p = 0.0487). However, the contrast-related criteria did not improve the pretherapeutic prediction of malignancy. Although contrast medium kinetics provided statistically significant new criteria for identifying malignant lymph nodes, differentiating between malignant and inflammatory changes in lymph nodes remains a diagnostic challenge.