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):33-6. doi: 10.4103/0255-0857.76521.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Bacteriological Techniques / methods*
  • Diagnosis, Differential
  • Female
  • Genitalia, Female / microbiology
  • Humans
  • Middle Aged
  • Mycoplasma Infections / diagnosis*
  • Mycoplasma Infections / microbiology
  • Mycoplasma genitalium / classification
  • Mycoplasma genitalium / genetics
  • Mycoplasma genitalium / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length*
  • Pregnancy
  • Pregnancy Complications, Infectious / diagnosis
  • Pregnancy Complications, Infectious / microbiology
  • Time Factors
  • Ureaplasma Infections / diagnosis*
  • Ureaplasma Infections / microbiology
  • Ureaplasma urealyticum / classification
  • Ureaplasma urealyticum / genetics
  • Ureaplasma urealyticum / isolation & purification*