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Test Code URPRP Ureaplasma species, Molecular Detection, PCR, Plasma

Useful For

Rapid, sensitive, and specific identification of Ureaplasma urealyticum and U parvum from plasma

Method Name

Real-Time Polymerase Chain Reaction (PCR) Using LightCycler and Fluorescent Resonance Energy Transfer (FRET)

Reporting Name

Ureaplasma PCR, P

Specimen Type

Plasma EDTA


Specimen Required


Collection Container/Tube:

Preferred: Lavender top (EDTA)

Acceptable: Royal blue top (EDTA), pink top (EDTA), or sterile vial containing EDTA-derived aliquot

Specimen Volume: 1 mL

Collection Instructions:

1. Spin down promptly. 

2. Send specimen in original tube (preferred). 

Additional Information: The high sensitivity of amplification by PCR requires the specimen to be processed in an environment in which contamination of the specimen by Ureaplasma DNA is not likely.


Specimen Minimum Volume

0.5 mL

Specimen Stability Information

Specimen Type Temperature Time
Plasma EDTA Refrigerated (preferred) 7 days
  Frozen  7 days

Reject Due To

Hemolysis

NA

Lipemia

NA

Icterus

NA

Other

Green-top (heparin) tube, anticoagulants other than EDTA

Clinical Information

Ureaplasma urealyticum and U parvum have been associated with a number of clinically significant infections, although their clinical significance may not always be clear as they are part of the normal genital flora. U urealyticum and U parvum have been associated with urethritis and epididymitis. They may cause upper urinary tract infection and they have been associated with infected renal stones. U urealyticum and U parvum may be isolated from amniotic fluid of women with preterm labor, premature rupture of membranes, spontaneous term labor, or chorioamnionitis. They may also cause neonatal infections, including meningoencephalitis and pneumonia. In addition, U urealyticum and U parvum have been reported to cause unusual infections, such as prosthetic joint infection and infections in transplant recipients.

 

Recently, U urealyticum and U parvum have been found to cause hyperammonemia in lung transplant recipients.(1) In lung transplant recipients with hyperammonemia, the ideal diagnostic specimen is a lower respiratory specimen (eg, bronchoalveolar lavage fluid), although U urealyticum and U parvum may also be detected in blood. Treatment directed against these organisms has resulted in resolution of hyperammonemia.

 

Culture of Ureaplasma species is laborious, requiring a high degree of technical skill and taking several days. PCR detection is sensitive, specific, and provides same-day results. In addition, PCR allows the differentiation of U urealyticum and U parvum, which is not easily accomplished with culture. PCR assay has replaced conventional culture for U urealyticum and U parvum at Mayo Medical Laboratories due to its speed and equivalent performance to culture.

Reference Values

Not applicable

Interpretation

A positive PCR result for the presence of a specific sequence found within the Ureaplasma urealyticum and U parvum ureC gene indicates the presence of U urealyticum or U parvum DNA in the specimen.

 

A negative PCR result indicates the absence of detectable U urealyticum and U parvum DNA in the specimen, but does not rule-out infection as false-negative results may occur due to inhibition of PCR, sequence variability underlying the primers and probes, or the presence of U urealyticum or U parvum in quantities below the limit of detection of the assay.

Cautions

Interfering substances may affect the accuracy of this assay; results should always be interpreted in conjunction with clinical and epidemiological findings.

 

Since Ureaplasma species may be part of the normal flora, results should be interpreted accordingly.

 

This test does not detect other mycoplasmas or ureaplasmas (including Mycoplasma pneumoniae, a common cause of community acquired pneumonia).

 

This test is not intended for medicolegal use.

Supportive Data

Validation included spiking studies for each Ureaplasma species. Spiking studies were carried out using 30 EDTA whole blood and plasma samples spiked with genomic DNA for U urealyticum and U parvum (as well as 10 naive specimens). Sensitivity and specificity was 100% for both targets.

Clinical Reference

1. Bharat A, Cunningham SA, Scott Budinger GR, Kreisel D, et al: Disseminated Ureaplasma infection as a cause of fatal hyperammonemia in humans. Sci Transl Med 2015;7(284):284re3

2. Stellrecht KA, Woron AM, Mishrik NG, Venezia RA: Comparison of multiplex PCR assay with culture detection of genital mycoplasmas. J Clin Microbiol 2004;42:1528-1533

3. Farrell JJ, Larson JA, Akeson JW, et al: Ureaplasma parvum prosthetic joint infection detected by PCR. J Clin Microbiol 2014;52:2248-2250

4. Waites KB, Taylor-Robinson D: Mycoplasma and Ureaplasma. In Manual of Clinical Microbiology. 11th edition. Edited by JH Jorgensen. ASM Press, Washington, DC, 2015, pp 1088-1105

5. Kenny GE: Genital mycoplasmas: Mycoplasma genitalium, Mycoplasma hominis, and Ureaplasma species. In Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases. Edited by GL Mandell, et al. Churchill Livingstone, New York, 2008

Method Description

This PCR method employs a target-specific detection system including primers, as well as fluorescent resonance energy transfer (FRET) hybridization probes designed for the ureC gene of Ureaplasma urealyticum and U parvum. The LightCycler instrument amplifies and monitors target nucleic acid sequences by fluorescence during PCR cycling. This is an automated PCR system that can rapidly detect amplified product development. The detection of amplified products is based on the FRET principle. For FRET product detection, a hybridization probe with a donor fluorophore, fluorescein, on the 3' end is excited by an external light source, which emits light that is absorbed by a second hybridization probe with an acceptor fluorophore, LC-Red 640, on the 5' end. The acceptor fluorophore then emits light of a different wavelength that is measured with a signal that is proportional to the amount of specific PCR product. The process is completed in a closed tube system and the melting temperature of the probes allows differentiation of U urealyticum from U parvum.(Cunningham SA, Mandrekar JN, Rosenblatt JE, Patel R: Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum. Int J Bacteriol 2013 Jan 30, doi: 10.1155/2013/168742)

Day(s) and Time(s) Performed

Monday through Friday

Analytic Time

3 days

Specimen Retention Time

7 days

Performing Laboratory

Mayo Medical Laboratories in Rochester

Test Classification

This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.

CPT Code Information

87798 x 2

LOINC Code Information

Test ID Test Order Name Order LOINC Value
URPRP Ureaplasma PCR, P 69934-8

 

Result ID Test Result Name Result LOINC Value
UPSRC Specimen Source 31208-2
44135 Ureaplasma urealyticum PCR, P 51988-4
44136 Ureaplasma parvum PCR, P 69933-0