Does the 1000 mL water leak test for medical gloves detect potential for virus penetration? Water leak and virus penetration were determined sequentially in hanging latex surgical gloves (4 brands) with 1000 ml of ΦX174-containing buffered saline (DPBS). Individual fingers (some punctured) were visually examined for water (DPBS) leaks for 2 minutes, then dipped into DPBS for 60 minutes for collection of virus that penetrated the latex barrier. Many punctured and a few non-punctured fingers leaked both water and virus. Some punctured fingers did not leak water nor virus. A few of the control and punctured fingers that did not leak water did allow very low-level virus penetration by 1 hour; 4–50 fold lower than virus penetration from barely-detectable water leaks. Thus, a few gloves with potential for very low-level virus penetration were not detected by the 1000 mL water leak test.