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 ORIGINAL ARTICLE
Year : 2011  |  Volume : 29  |  Issue : 1  |  Page : 33-36

Simultaneous and rapid differential diagnosis of Mycoplasma genitalium and Ureaplasma urealyticum based on a polymerase chain reaction-restriction fragment length polymorphism


1 Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
2 Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
3 Cell and Molecular Biology Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran

Correspondence Address:
R Mirnejad
Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran
Iran
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Source of Support: Iran University of medical science, Conflict of Interest: None


DOI: 10.4103/0255-0857.76521

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Objectives: The aim of this investigation was to simultaneously detect and differentiate Mycoplasma genitalium and Ureaplasma urealyticum in female patients suffering from genital complications by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Materials and Methods : Genital swabs were taken from 210 patients. They were transported to the laboratory in phosphate-buffered saline. For PCR, samples were analysed with genus-specific MyUu-R and MyUu-F primers. This primer set, which was originally designed in our laboratory, amplified a 465 bp fragment (M. genitalium) and a 559 bp fragment (U. urealyticum). Samples containing a band of the expected sizes for the Mycoplasma strains were subjected to digestion with a restriction endonuclease enzyme of TaqI and Cac8I. Results: Of the 210 samples, a total of 100 (47.6%) samples were found to be positive for Mycoplasmas (seven M. genitalium isolates, 3.3%; and 89 U. urealyticum isolates, 42.4%), and coinfections with both species were detected in four samples (1.9%). The PCR-RFLP results showed that M. genitalium and U. urealyticum are different by enzyme patterns. Conclusion: PCR-RFLP offers a rapid and easily applicable protocol to simultaneous detection and differentiation of M. genitalium and U. urealyticum from clinical samples when specific primers and restriction enzymes are used.






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2004 - Indian Journal of Medical Microbiology
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