Introduction Examination gloves manufactured from natural latex have been the predominant glove choice to date in dental practice. However, concerns over hypersensitivity have resulted in the use of alternatives such as nitrile gloves. The aim of the current study was to assess the incidence of pre-existing pinhole defects in nitrile examination gloves. Methods Air inflation, followed by water submersion, was used to assess the incidence of pre-existing pinhole defects in five nitrile and two latex glove types. The gloves were filled with a constant volume of air and submerged in 3 litres of water for 10 seconds while being observed for air bubbles which would indicate pinhole defects. The position and number of pinholes were noted for 100 gloves of each type investigated. Results The incidence of pre-existing pinholes for latex gloves was 0% for the non-sterile surgical latex glove type and 3% for the powdered latex examination glove type, with pinholes located on the thumb, middle finger and ring finger. Of the nitrile gloves evaluated, three types were assessed to have no pre-existing pinhole defects. One type had a 2% incidence of pre-existing pinhole defects -one pinhole located on the thumb region of the glove and one on the ring finger portion of the glove. The fifth nitrile glove type had one pre-existing pinhole defect located on the middle finger. Significance All glove types examined met the European Standard (EN 455-1) and there was no statistically significant difference between glove types. However, the nitrile gloves generally exhibited less pre-existing pinhole defects than the latex examination gloves.