: Currently, the Meares-Stamey 4-glass and the 2-glass tests are used for diagnosing chronic prostatitis subtypes. Both tests include prostatic massage. Failure to extract prostatic secretions-for any reason-can result in a non-diagnostic test. Evidence from everyday practice and studies shows that expressed prostatic secretions are successfully recovered in less than 50% of the examined patients, and an important number of post-massage urine samples are missing prostatic secretions. This study evaluated a simpler test, the 3-glass (pre-ejaculation, ejaculation, and post-ejaculation) test. We compared it with the 4-glass and the 2-glass tests to detect inflammation and bacteria in men with chronic prostatitis symptoms. The study population included patients with chronic prostatitis symptoms. Subjects were assigned in each visit to undergo either the 4-glass, the 2-glass test, or the 3-glass test. The comparison among the three tests was based on the percentage of bacterial detection, the percentage of false-negative diagnoses, and the percentage of shifts among chronic prostatitis subtypes in the follow-up visits of recurrent patients. A total of 157 patients were finally evaluated. Fifty-nine (59) patients underwent the 4-glass test (Group A), sixty-seven (67) underwent the 3-glass test (Group B), and thirty-one (31) underwent the 2-glass test (Group C). No statistically significant differences in the comparisons above were found. : A comparison of the three diagnostic tests showed equivalence of the total ejaculate culture-based 3-glass test to the conventional prostatic secretion culture-based tests.