Detection of Ureaplasma urealyticum by Polymerase Chain Reaction in the Urogenital Tract of Adults, in Amniotic Fluid, and in the Respiratory Tract of Newborns

Abstract

On the basis of the nucleotide sequence of the urease genes of Ureaplasma urealyticum serotype 8, polymerase chain reaction (PCR) primers were selected, evaluated for specificity and sensitivity, and tested for their ability to detect U. urealyticum in the adult urogenital tract, amniotic fluid, and endotracheal aspirates of newborns. All 14 reference serotypes of U. urealyticum were detected with equal sensitivity (1-10 cfu), whereas multiple strains of 12 other mycoplasma species found in humans as well as eukaryotic DNA were not detected. A total of 638 clinical specimens was evaluated. Results indicate that PCR is equal to if not more sensitive than culture for detection of U. urealyticum. Faster detection of U. urealyticum by PCR (< 24 h) compared to culture (2-5 days) will be particularly important in management of very low birth-weight infants in whom this organism has been shown to be a significant cause of meningitis, respiratory disease, and death. This method of detection will also be helpful in further determining the role of this organism in intraamniotic infection and premature birth.

Detection and biovar discrimination of Ureaplasma urealyticum colonization in preterm neonates under ventilation and correlation with bronchopulmonary dysplasia

“…This is in agreement with the percentage of colonization of U. urealyticum in neonatal respiratory specimens reported in other studies from 3 to 23% (Blanchard et al, 1993;van Waarde et al, 1997;Payne et al, 1993;Jonsson et al, 1994;Mohagheghi et al, 2013). Higher rates were reported by Nelson et al (1998) (2000) (55%) that could be explained by larger sample size, or testing multiple specimens from enrolled patients.…”

“…urealyticum is among the less frequently diagnosed respiratory pathogens in a clinical environment, mainly because of the lack of standardized and specific diagnostic tests (Blanchard et al, 1993). Routine bacterial cultures are not sufficient to recover Ureaplasma species.…”

Patterns of Colonization WithUreaplasma urealyticumDuring Neonatal Intensive Care Unit Hospitalizations of Very Low Birth Weight Infants and the Development of Chronic Lung Disease

“…35 Broths were thawed and centrifuged at 12 000 g at 4°C for 20 minutes. The supernatant was discarded; the pellet was resuspended in 50 L solution A (TRIS HCL pH 8.3 10 mM, KCL 100 mM, MgCl 2 2.5 mM) and an equal volume of solution B (TRIS HCL 8.3 20 mM, Tween 2%, MgCl 2 5 mM, Trition-X 2%, Proteinase K [GIBCO, Gaithersburg, MD, 0.5 mg/mL], Proteinase K buffer [TRIS HCL 7.5 10 mM, CaCl 2 20 mM, Glycerol 50%]) and incubated at 60°C for 60 minutes then heated to 100°C for 10 minutes.…”

Comparison of Culture With the Polymerase Chain Reaction for Detection of Gennital Mycoplasma

“…Specific oligonucleotide primers were used for U.urealyticum and M. hominis (Table 1). These were chosen in published nucleotide sequences: in the urease gene for U.urealyticum (18) and in the 16S rRNA gene for M. hominis (19). Amplification was performed in 50µl of reaction mixture containing 10µl of 10× PCR buffer, 2.5 mM Mgcl 2 , 200µM dNTP, 1.25 units of Taq polymerase, 20pmol of each primer and 7µl of sample DNA.…”