AbstractBackgroundUreaplasmaspp.are associated with various infectious diseases in females, but there is still limited evidence regarding whether they are related to nonspecific cervicitis. The aim of this study was to develop and evaluate a digital droplet PCR (ddPCR) assay for the detection and quantification ofUreaplasmaspp. in cervical swabs.MethodsA total of 267 non-specific cervicitis (NSC) patients and 195 asymptomatic females were included in this study. We produced standard curves forUreaplasmaspp.to evaluate the analytical performance of the ddPCR assay. Then, we detected and quantified the bacterial load ofUreaplasmaspp. in cervical swabs.ResultsThe prevalences ofU. parvumwere 37.8% (101/267) and 29.7% (58/195), U. urealyticumwere 9.0% (24/267) and 8.7% (17/195) in the NSC group and control group, respectively. In addition, the median copy number ofU. parvumwas 2.5 × 10⁴copies/ml (n = 101) in the NSC group and 9.2 × 10³copies/ml (n = 58) in the control group. TheU. parvumload in the NSC group was significantly higher than that in the asymptomatic individuals (P < 0.001). whereas the median load ofU. urealyticumwas 8.4 × 10³copies/ml (n = 24) and 1.4 × 10³(n = 17) copies/ml in the two groups, respectively, , the difference was not statistically significant (P = 0.450).ConclusionsOur study is the first to develop a droplet digital PCR (ddPCR) method for the detection and quantification ofUreaplasmaspp. in clinical samples, and the method has excellent analytical performance and a wide range of clinical application prospects.