The prolactin test measures the levels of prolactin, a hormone produced by the pituitary gland, and is used to diagnose various conditions in both males and females. In females, the test is conducted for issues like menstrual irregularities and lactation failure, while in males, it helps in assessing decreased sexual drive and erectile dysfunction. Normal prolactin levels vary by gender and pregnancy status, and specific pre-test instructions must be followed for accurate results.