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Updated: Aug 30, 2026
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
[Development of a multiple PCR method for the identification of Ureaplasma parvum and Ureaplasma urealyticum]
C Fernández Molina1, M A Latino, Y Zamora Martínez
1Instituto de Medicina Tropical Pedro Kourí, A.P. 601, Mariano 13, La Lisa, Ciudad de La Habana, Cuba. carmen@ipk.sld.cu
Insights
A new Multiplex-PCR method accurately identifies Ureaplasma parvum and Ureaplasma urealyticum. This technique is crucial for diagnosing Ureaplasma-associated disorders in newborns, women, and men.
Area of Science:
- Microbiology
- Molecular Biology
- Clinical Diagnostics
Background:
- Ureaplasma parvum and Ureaplasma urealyticum are linked to various health issues, particularly in underweight newborns.
- Accurate identification of Ureaplasma species is essential for effective clinical management.
- Existing diagnostic methods may require enhancement for precise species differentiation.
Purpose of the Study:
- To develop and validate a novel Multiplex-PCR method for differentiating Ureaplasma parvum and Ureaplasma urealyticum.
- To assess the prevalence of U. parvum and U. urealyticum in clinical samples from different patient groups.
Main Methods:
- Development of a Multiplex-PCR assay utilizing specific primers (UPS-UPSA for U. parvum, UUS2-UUA2 for U. urealyticum).
- Application of the Multiplex-PCR method to identify Ureaplasma species in clinical samples previously identified by the "MYCOFAST Evolution-2" Kit.
Main Results:
- The Multiplex-PCR method successfully identified Ureaplasma species in 56 samples from newborns, 76 from women, and 63 from men.
- In newborns, 70% were U. parvum and 30% U. urealyticum.
- In women, 83% were U. parvum and 17% U. urealyticum; in men, 76% were U. parvum and 24% U. urealyticum.
Conclusions:
- The developed Multiplex-PCR method offers high specificity for identifying clinically relevant Ureaplasma species.
- This molecular technique provides a reliable tool for diagnosing Ureaplasma infections across diverse patient populations.
- The findings highlight the predominant presence of Ureaplasma parvum in the studied clinical samples.
Abstract:
Ureaplasma parvum and Ureaplasma urealyticum, also known as biovar parvum and biovar T960, respectively, could be associated with several disorders in men, women, and mainly, in newborn children with under weight. Several methods have been developed in order to identify the species or biovars of ureaplasmas. We developed a Multiplex-PCR method using the UPS-UPSA and UUS2-UUA2 primers, specific for U. parvum and U. urealyticum, respectively. This Multiplex-PCR method was used to identify cultures of clinical positive samples to Ureaplasma spp. by the "MYCOFAST Evolution-2" Kit. Of 56 positive cultures to Ureaplasma spp. from newborn children, 70% were U. parvum and 30% U. urealyticum; in 76 positive samples in women, 83% corresponded to U. parvum and 17% to U. urealyticum, while in 63 positive samples of men, 76% identified U. parvum and 24% U. urealyticum. The PCR-multiplex method showed specificity for the identification of the biovars or species of ureaplasmas of clinical interest.
