Background: Ureaplasma parvum and Ureaplasma urealyticum are commonly found in the cervix of women with non-chlamydial and non-gonococcal cervicitis or non-specific cervicitis (NSC). However their contribution to the aetiology of NSC is controversial. Methods: U. parvum and U. urealyticum were identified and quantified in cervical swabs collected from 155 women with NSC and 312 controls without NSC, using real-time PCR. The relative bacterial quantification was then calculated using the Ureaplasma copy number divided by the number of host cells; this is important for the correction of bias linked to the number of cells harvested in different swabs. Results: Ureaplasma was detected in 58.7% (91/155) of NSC patients: U. parvum in 30.3%, U. urealyticum in 16.1%, and mixed infection in 12.3%. It was also detected in 54.5% (170/312) of controls: U. parvum in 33.0%, U. urealyticum in 11.5%, and mixed infection in 9.9%. There were no significant differences for U. parvum, U. urealyticum, or mixed infection between the 2 groups (p > 0.05). However, both biovars were present at higher concentrations in NSC patients than in controls (p < 0.05). Using >10 copies/1000 cells as a reference, the positive rate of U. parvum in NSC patients was 16.1%, significantly higher than that in controls at 5.1% (relative risk 3.145, p < 0.05); positive rates of U. urealyticum in NSC patients and controls were 28.4% and 8.7%, respectively, with a statistically significant difference (relative risk 3.131, p < 0.05). Conclusions: Ureaplasma can adhere to host cells, colonize, internalize, and subsequently produce pathological lesions. A high density of Ureaplasma in the cervix may be associated with the aetiology of NSC.