OBJECTIVE: To prospectively determine the prevalence of testicular microlithiasis in symptomatic patients who were referred for scrotal ultrasound examination and to evaluate the possible association of testicular microlithiasis with testicular cancer and other conditions such as cryptorchidism or history of ascending testis. MATERIALS AND METHODS: 391 men who were referred to our institutions between July 2002 and May 2005 for any type of symptoms from the testicles, underwent physical and scrotal ultrasound examination. The presence of testicular microlithiasis, the number of lesions and the involvement of both testicles in relation to the symptoms as well as the coexistence of other lesions were studied. RESULTS: Eighteen (4.6%) of 391 men enrolled into the study had testicular microlithiasis. Two out of the eighteen patients (11%) had concomitant testicular cancer, which was confirmed by pathological evaluation of the orchidectomy specimen. One of the patients with testicular microlithiasis presented a rising in biochemical tumor markers (LDH, and HCG) and underwent orchidectomy one year later. Five of the remaining 373 (1.3%) patients without microlithiasis were diagnosed with testicular cancer. Thirty six men reported having a history of ascending testis, but none of them was found with testicular cancer. Two cases of testicular torsion in a cryptorchid position had testicular microlithiasis, but the orchidectomy specimen (after surgery) was negative for testicular cancer. The correlation between testicular cancer and testicular microlithiasis found in our study was statistically significant (p < 0.05). CONCLUSION: There seems to be an association between testicular microlithiasis and testicular cancer.