Parvovirus B19, Molecular Detection, PCR, Plasma
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
Diagnosing erythrovirus B19 (parvovirus) infection
Real-Time Polymerase Chain Reaction (PCR)/DNA Probe Hybridization
(PCR is utilized pursuant to a license agreement with Roche Molecular Systems, Inc.)
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
Parvovirus B19 PCR, P
Human Parvovirus B19
Parvovirus B19 PCR
Human Parvovirus B19
Parvovirus B19 PCR
Specimen Type Describes the specimen type needed for testing
Specimen Required Defines the optimal specimen. This field describes the type of specimen required to perform the test and the preferred volume to complete testing. The volume allows automated processing, fastest throughput and, when indicated, repeat or reflex testing.
Container/Tube: Lavender top (EDTA)
Specimen Volume: 0.5 mL
Forms: If not ordering electronically, submit a Microbiology Request Form (Supply T244) with the specimen.
Specimen Minimum Volume Defines the amount of specimen required to perform an assay once, including instrument and container dead space. Submitting the minimum specimen volume makes it impossible to repeat the test or perform confirmatory or perform reflex testing. In some situations, a minimum specimen volume may result in a QNS (quantity not sufficient) result, requiring a second specimen to be collected.
Mild OK; Gross reject
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
|Plasma EDTA||Refrigerated (preferred)||7 days|
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
B19, previously classified as a parvovirus, is now in the genus erythrovirus based on preferential replication of this virus in erythroid progenitor cells.(1) Infection with B19 occurs early in life and the virus is transmitted by respiratory secretion and occasionally by blood products. Antibody prevalence ranges from 2% to 15% in early adults.(1)
B19 may result in an asymptomatic infection or produce a wide spectrum of disease ranging from erythema infections (slapped cheek syndrome or fifth disease) in children to arthropathy, severe anemia, and systemic manifestations involving the central nervous system, heart, and liver depending on the immune competence of the host. (2,3) Infection with B19 in pregnant women may cause hydrops fetalis, congenital anemia, abortion, or stillbirth of the fetus.(4) B19 is also the causative agent of persistent anemia usually, but not exclusively, in immunocompromised patients, transplant patients, and infants. The deficiencies of appropriate immune responses to B19 impair viral elimination in virus, which results in enlargement of B19-infected erythroid-lineage cells.(5,6)
Most acute infections with B19 are diagnosed in the laboratory by serologically detecting IgG and IgM class antibodies with enzyme-linked immunosorbent assay testing.
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
The presence of erythrovirus B19 DNA by PCR indicates infection with this virus.
The absence of erythrovirus B19 DNA by PCR indicates the lack of infection with this virus (see Cautions).
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
A negative result does not necessarily indicate the absence of B19 infection. False-negative results may be due to suppression of virus replication to levels below the detection threshold, or to inhibitory substances that may be present in the specimens.
Inadequate specimen collection or storage may invalidate test results.
The following data supports the use of this assay for clinical testing.
Accuracy/Diagnostic Sensitivity and Specificity:
Results from this real-time 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 as the gold standard, the Diagnostic Sensitivity and Specificity for detection of Parvovirus B19 was 97%.
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 (10-20 targets/mcL). The 30 spiked specimens (45 bloods) were run in a blinded manner along with 30 negative (non-spiked) specimens (45 bloods). 97% to 100% of the spiked specimens were positive and 100% of the non-spiked specimens were negative.
Analytical Sensitivity/Limit of Detection:
The lower limit of detection (LoD) of this assay is 10 to 20 targets/microL in sample matrix.
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.
Inter-assay precision was 100% and intra-assay precision was 97%.
Although the reference range is typically "negative" for this assay, this assay may detect viremia in asymptomatic individuals. 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 is not to be used to screen asymptomatic patients.
This is a qualitative assay, and results are reported as either negative or positive for targeted parvovirus B19.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Heegaard ED, Brown KE: Human parvovirus B19. Clin Microbiol Ref 2002;15:485-505
2. Bultmann BD, Klingel K, Soltar K, et al: Fatal parvovirus B19 associated myocarditis clinically mimicking ischemic heart disease: an endothelial cell-mediated disease. Hum Pathol 2003;34:92-95
3. Rerolle JP, Helal I, Morelon E: Parvovirus B19 infection after renal transplantation. Nephrologie 2003;24:309-315
4. Chisaka H, Morita E, Yaegashi N: Parvovirus B19 and the pathogenesis of anaemia. Rev Med Virol 2003;16:347-359
5. Goto H, Ishida A, Fujii H, et al: Successful bone marrow transplantation for severe aplastic anemia in a patient with persistent human parvovirus B19 infection. Int J Hematol 2004;79(4):384-386
Method Description Describes how the test is performed and provides a method-specific reference
Viral DNA is extracted from 0.2 mL of specimen by the MagNA Pure automated instrument (Roche Applied Science). LightCycler 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 degrees C, the temperature in the thermal chamber is slowly raised to 80 degrees 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;78:1-2;191-198)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday; Varies
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Same day/1 day
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test
Test Classification Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer's instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR), Investigation Use Only (IUO) product, or a Research Use Only (RUO) product.
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 Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Medical Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.
LOINC® Code Information Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the result codes returned for this test or profile.
|Result ID||Reporting Name||LOINC Code|
|56075||Parvovirus B19 By Rapid PCR||9572-9|