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Simultaneous and rapid differential diagnosis of Mycoplasma genitalium and Ureaplasma urealyticum based on a polymerase chain reaction-restriction fragment length polymorphism.
Indian J Med Microbiol ; 2011 Jan-Mar; 29(1): 33-36
Article in English | IMSEAR | ID: sea-143772
ABSTRACT

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.
Subject(s)

Full text: Available Index: IMSEAR (South-East Asia) Main subject: Pregnancy Complications, Infectious / Time Factors / Polymorphism, Restriction Fragment Length / Aged / Female / Humans / Pregnancy / Polymerase Chain Reaction / Bacteriological Techniques / Ureaplasma urealyticum Type of study: Diagnostic study / Practice guideline Language: English Journal: Indian J Med Microbiol Journal subject: Microbiology Year: 2011 Type: Article

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Full text: Available Index: IMSEAR (South-East Asia) Main subject: Pregnancy Complications, Infectious / Time Factors / Polymorphism, Restriction Fragment Length / Aged / Female / Humans / Pregnancy / Polymerase Chain Reaction / Bacteriological Techniques / Ureaplasma urealyticum Type of study: Diagnostic study / Practice guideline Language: English Journal: Indian J Med Microbiol Journal subject: Microbiology Year: 2011 Type: Article