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HelpTest Code LAB1231753 Parvovirus B19, Molecular Detection, PCR, Plasma
Performing Laboratory
Mayo Clinic Laboratories in Rochester
Reporting Name
Parvovirus B19 PCR, PSpecimen Type
Plasma EDTASpecimen Required
Supplies: Sarstedt Aliquot Tube 5 mL (T914)
Collection Container/Tube: Lavender top (EDTA)
Submission Container/Tube: Plastic vial
Specimen Volume: 0.5 mL
Collection Instructions: Centrifuge and aliquot plasma into a plastic vial.
Reject Due To
| Gross hemolysis | Reject |
Specimen Stability Information
| Specimen Type | Temperature | Time |
|---|---|---|
| Plasma EDTA | Refrigerated (preferred) | 7 days |
| Frozen | 7 days |
Specimen Minimum Volume
0.3 mL
Day(s) Performed
Monday through Friday
Specimen Retention Time
1 weekReport Available
Same day/1 to 5 daysReference Values
Negative
Useful For
Diagnosing parvovirus B19 infection in plasma specimens
CPT Code Information
87798
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| PARVP | Parvovirus B19 PCR, P | 9571-1 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| 56075 | Parvovirus B19 By Rapid PCR | 9571-1 |
| SS008 | Source | 31208-2 |
Method Description
Viral DNA is extracted from 0.2 mL of specimen by the MagNA Pure automated instrument (Roche Applied Science). LightCycler polymerase chain reaction (PCR) primers and probes detect target B19 DNA (nonstructural protein). The LightCycler instrument amplifies and monitors by fluorescence the development of target nucleic acid sequences after the annealing step during PCR cycling. This automated PCR system can rapidly detect (30-40 minutes) amplicon development through stringent air-controlled temperature cycling in capillary cuvettes. The detection of amplified products is based on the fluorescence resonance energy transfer (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 and emits light that is absorbed by a second hybridization probe with an acceptor fluorophore, LC-Red 640, at the 5' end. The acceptor fluorophore then emits a light of a different wavelength that can be measured with a signal that is proportional to the amount of specific PCR product. Melting curve analysis is performed following PCR amplification. Starting at 45° C, the temperature in the thermal chamber is slowly raised to 80° C and the fluorescence is measured at frequent intervals. Analysis of the PCR amplification and the probe melting curves is accomplished through the use of LightCycler software.(Soares RM, Durigon El, Bersano JG, Richtzenhain LG: Detection of porcine parvovirus DNA by the polymerase chain reaction assay using primers to the highly conserved nonstructural protein gene, NS-1. J Virol Methods. 1999 Mar;78(1-2):191-198)
Supportive Data
The following data supports the use of this assay for clinical testing.
Accuracy/Diagnostic Sensitivity and Specificity:
Results from this real-time polymerase chain reaction (PCR) assay on the LightCycler (LC PCR) were compared to a Centers for Disease Control and Prevention (CDC) PCR-based assay on tissue biopsy specimens of temporal artery. Using the CDC PCR method as the gold standard, the diagnostic sensitivity and specificity for detection of parvovirus B19 was 97%.
Supplemental Data:
To supplement the above data, 30 negative cerebrospinal fluid, body fluids, and tissues and 45 negative blood specimens were spiked with parvovirus B19-positive control plasmid at the limit of detection (LOD) (10-20 targets/microliter). The 30 spiked specimens (45 bloods) were run in a blinded manner along with 30 negative (nonspiked) specimens (45 bloods). Results showed 97% to 100% of the spiked specimens were positive and 100% of the nonspiked specimens were negative.
Analytical Sensitivity/LOD:
The lower LOD of this assay is 10 to 20 targets/microliter in sample matrix.
Analytical Specificity:
No PCR signal was obtained with extracts of 11 viral and bacterial isolates that may cause symptoms similar to infection with parvovirus, including herpes simplex virus, varicella-zoster virus, cytomegalovirus, human herpesvirus-6, -7, and -8.
Precision:
Interassay precision was 100% and intra-assay precision was 97%.
Reference Range:
Although the reference range is typically "negative" for this assay, this assay may detect viremia in asymptomatic individuals or remnant viral nucleic acid. However, this assay is only to be used for patients with a clinical history and symptoms consistent with parvovirus B19 infection and must be interpreted in context of clinical picture. This test should not be used to screen asymptomatic patients.
Reportable Range:
This is a qualitative assay, and results are reported as either negative or positive for targeted parvovirus B19.
Forms
If not ordering electronically, complete, print, and send 1 of the following forms with the specimen:
-Kidney Transplant Test Request
-Microbiology Test Request (T244)