title: Simultaneous and rapid differential diagnosis of Mycoplasma genitalium and Ureaplasma urealyticum based on a polymerase chain reaction-restriction fragment length polymorphism.
abstract: OBJECTIVE
  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).
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Baqiyatallah University of Medical Sciences, Tehran, Iran. rmirnejadreza@yahoo.com
authors.forename: R
authors.initials: R
authors.lastname: Mirnejad
authors.forename: N
authors.initials: N
authors.lastname: Amirmozafari
authors.forename: B
authors.initials: B
authors.lastname: Kazemi
mesh_terms: D000328:Adult; D000368:Aged; D001431:Bacteriological Techniques / Q000379:methods*; D003937:Diagnosis, Differential; D005260:Female; D005836:Genitalia, Female / Q000382:microbiology; D006801:Humans; D008875:Middle Aged; D009175:Mycoplasma Infections / Q000175:diagnosis* / Q000382:microbiology; D045704:Mycoplasma genitalium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000175:diagnosis / Q000382:microbiology; D013997:Time Factors; D016869:Ureaplasma Infections / Q000175:diagnosis* / Q000382:microbiology; D016990:Ureaplasma urealyticum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*