Adenovirus, Molecular Detection, PCR, Plasma
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
As an aid in diagnosing adenovirus infections.
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
Adenovirus PCR, P
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: 1 mL
Collection Instructions: Spin down promptly.
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||7 days|
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Human adenoviruses cause a variety of diseases including pneumonia, cystitis, conjunctivitis, diarrhea, hepatitis, myocarditis, and encephalitis. In humans, adenoviruses have been recovered from almost every organ system. Infections can occur at any time of the year and in all age groups. Currently, there are 51 adenovirus serotypes that have been grouped into 6 separate subgenera.
Culture is the gold standard for the diagnosis for adenovirus infection; however, it can take up to 3 weeks to achieve culture results (Mayo's shell vial culture provides more rapid results, reported at 2 and 5 days). Serological tests have faster turnaround times, but can be less sensitive compared to culture. PCR offers a rapid, specific, and sensitive means of diagnosis by detecting adenovirus DNA.
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.
A positive result indicates the presence of adenoviruses.
A negative result does not rule out the presence of adenoviruses because organisms may be present at levels below the detection limits of this assay.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
Test results should be used as an aid in diagnosis and should not be considered diagnostic in themselves.
Although the reference range is generally considered to be "Negative" for this assay, adenovirus DNA may be detected from asymptomatic individuals in certain settings. This assay should only be used to test patients with clinical history and symptoms consistent with adenovirus disease, and is not used to screen healthy patients.
The following support the use of this assay for clinical testing.
Accuracy/Diagnostic Sensitivity and Specificity:
A study of 715 clinical specimens compared shell vial culture and this PCR assay. Included in the study were 286 swab specimens (nasal, throat, rectal, skin), 49 eye specimens, 221 respiratory specimens (bronchial washings, sputa, bronchioalveolar lavage, tracheal secretions), 55 fresh tissue specimens, 72 stools, and 27 body fluids/other specimens. Specimens were inoculated into culture tubes and examined for cytopathic effects over a period of 14 days, and subsequently assayed with this LightCycler assay. Comparison of cell culture with LC PCR yielded the following: total specimens positive by LC PCR was 60 [(stool=9), (respiratory=4), (tissue=4), (swabs=24), (eye specimens=14), and (urine=4)] and total specimens by culture were 52 [(stool= 8), (respiratory=3), (tissue=3), (swabs=23), (eye specimens=13), and (urine=2)]. Of the 60 total positive specimens, PCR produced a 13.5% increased rate of detection of adenovirus compared with culture. Analytical sensitivity was assessed by testing dilutions (in triplicate) of the plasmid control down to a level corresponding to 1 target/mL. The limit of reproducible detection was determined to be 10 targets/mL. Additionally, the sensitivity of plasma was known to be > or =90% at the concentration of 10 targets/L. This assay detected all 51 serotypes of Adenovirus tested.
Supplemental Data (Spiking Studies):
To supplement the above data, 30 negative samples of various types (CSF, ocular, respiratory, stool, urine, and plasma) were spiked with adenovirus positive control plasmid at the limit of detection (approximately 10 targets/L). The 30 spiked specimens were run in a blinded manner with 30 negative (non-spiked) specimens. 100% of the spiked specimens were positive and 100% of the non-spiked specimens were negative.
Analytical Sensitivity/Limit of Detection (LoD):
The lower limit of detection of this assay is 10 targets/L in specimen matrix.
No PCR signal was obtained from extracts of 150 bacterial, viral, parasitic, and fungal isolates that could cause similar disease or could be found as normal flora in sites normally tested for this organism.
Inter-assay precision was 100% and intra-assay precision was 100%.
The reference range for this assay is "Negative."
This is a qualitative assay and results are reported as negative or positive for targeted adenovirus DNA.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Ebner K, Pinsker W, Lion T: Comparative sequence analysis of the hexon gene in the entire spectrum of human adenovirus serotypes: phylogenetic, taxonomic, and clinical implications. J Virol 2005;79:12635-12642
2. Ebner K, Suda M, Watzinger F, Lion T: Molecular detection and quantitative analysis of the entire spectrum of human adenoviruses by a two-reaction real-time PCR assay. J Clin Microbiol 2005;43:3049-3053
3. Jothikumar N, Cromeans TL, Hill VR, et al: Quantitative real-time PCR assays for the detection of human adenoviruses and identification of serotypes 40 and 41. Appl Environ Microbiol 2005;71:3131-3136
4. Robinson C, Echavarria M: Adenovirus. In Manual of Clinical Microbiology. Edited by PR Murray, EJ Baron, JH, et al: Washington, DC, ASM Press, 2007, pp 1589-1600
5. Thavagnanam S, Christie SN, Doherty GM, et al: Respiratory viral infection in lower airways of asymptomatic children. Acta Paediatr. Mar;99(3):394-398
6. Kaneko H, Maruko I, Iida T, et al: The possibility of human adenovirus detection in the conjunctiva in asymptomatic cases during a nosocomial infection. Cornea. Jun 2008;27(5):527-530
Method Description Describes how the test is performed and provides a method-specific reference
Respiratory, swabs, stools, tissues, plasma, and urine samples were processed according to specimen source. Viral nucleic acid is extracted by the MagNA Pure automated instrument (Roche Applied Science). Primers and fluorescence resonance energy transfer (FRET) probes target a relatively conserved 185-base pair region of the adenovirus penton gene. The LightCycler instrument (Roche Applied Science) amplifies and monitors the development of target nucleic acid sequences after the annealing step during PCR cycling. This automated PCR system rapidly detects amplicon development through stringent air-controlled temperature cycling in capillary cuvettes. 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 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. (Unpublished Mayo method).
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, Wednesday, Friday; 6 a.m.
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.
Monday through Thursday: 2 days Friday, Saturday: 3 days
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|