Related Experiment Video
Updated: Aug 20, 2026
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Development of real-time PCR for the differential detection and quantification of Ureaplasma urealyticum and
K Mallard1, K Schopfer, T Bodmer
1Institute for Infectious Diseases, University of Berne, Friedbuehlstrasse 51, CH-3010 Berne, Switzerland.
Abstract:
Ureaplasma parvum and Ureaplasma urealyticum are recently recognized species of the genus Ureaplasma. In humans, Ureaplasma spp. can be found on mucosal surfaces, primarily in the respiratory and urogenital tracts. They have been implicated in various human diseases such as nongonococcal urethritis, intrauterine infections in association with adverse pregnancy outcome and fetal morbidity, and pneumonitis in immunocompromised hosts. We have developed two quantitative real-time PCR assays to differentially detect U. parvum and U. urealyticum. Based upon the sequence information of the urease gene (ureB), we designed two TaqMan primer and probe combinations specific for U. parvum and U. urealyticum, respectively. The assays did not react with nucleic acid preparations from 16 bacterial species commonly encountered in relevant clinical specimens, including seven urease-producing species. Each assay had a detection limit of approximately five copies per reaction of the respective gene target. The results suggest that these assays are both sensitive and specific for U. parvum and U. urealyticum. Further investigation of both assays using clinical specimens is appropriate.
Insights
New real-time PCR assays can now differentiate Ureaplasma parvum and Ureaplasma urealyticum. These sensitive and specific methods aid in diagnosing infections caused by these common mucosal bacteria.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Ureaplasma parvum and Ureaprealyticum are recognized species within the Ureaplasma genus.
- These bacteria colonize mucosal surfaces, particularly in the respiratory and urogenital tracts.
- Ureaplasma species are linked to conditions like nongonococcal urethritis, adverse pregnancy outcomes, and pneumonitis in immunocompromised individuals.
Purpose of the Study:
- To develop and validate quantitative real-time PCR assays for the differential detection of Ureaplasma parvum and Ureaplasma urealyticum.
Main Methods:
- TaqMan primer and probe combinations were designed based on the urease gene (ureB) sequence.
- Two distinct quantitative real-time PCR assays were developed, one for U. parvum and one for U. urealyticum.
- Assay specificity was tested against 16 common bacterial species, including urease-producing ones.
Main Results:
- Both developed assays demonstrated high specificity, showing no cross-reactivity with other tested bacterial species.
- The detection limit for each assay was approximately five gene copies per reaction, indicating high sensitivity.
- The assays successfully differentiated between U. parvum and U. urealyticum.
Conclusions:
- The developed quantitative real-time PCR assays are sensitive and specific for detecting Ureaplasma parvum and Ureaplasma urealyticum.
- These assays provide a valuable tool for accurate diagnosis and further research into Ureaplasma-associated conditions.
- Clinical validation of these assays is warranted to assess their utility in diagnosing human infections.
