Test Catalog

Test Id : ENTP

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Enterovirus, Molecular Detection, PCR, Plasma

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Overview

Test Catalog

Useful For
Suggests clinical disorders or settings where the test may be helpful

Aiding in diagnosing enterovirus infections using plasma specimens

 

This test should not be used to screen asymptomatic patients.

Method Name
A short description of the method used to perform the test

Real-Time Polymerase Chain Reaction (PCR)/RNA Probe Hybridization

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

Enterovirus PCR, P

Aliases
Lists additional common names for a test, as an aid in searching

Coxsackievirus

Coxsackievirus A

Coxsackievirus B

Echovirus

Enterovirus

Enterovirus (Coxsackievirus and Echovirus)

Hand, Foot, and Mouth Disease

Specimen Type
Describes the specimen type validated for testing

Plasma EDTA

Ordering Guidance

This test will detect enterovirus but will not differentiate viruses in this family or provide serotyping information.

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Submit a raw clinical sample (not a culture isolate) for enterovirus polymerase chain reaction .

 

Collection Container/Tube: Lavender top (EDTA)

Submission Container/Tube: Screw-capped, sterile container

Specimen Volume: 1 mL

Collection Instructions: Centrifuge blood collection tube and aliquot plasma into plastic vial.

Forms

If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the testing laboratory. The minimum volume is sufficient for one attempt at testing.

0.3 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis Reject

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

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

Useful For
Suggests clinical disorders or settings where the test may be helpful

Aiding in diagnosing enterovirus infections using plasma specimens

 

This test should not be used to screen asymptomatic patients.

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Enteroviruses are positive-sense RNA viruses in the Picornaviridae family. These viruses were initially classified by serotype as polioviruses (3 types), echoviruses (31 types, including types 22 and 23, which are now classified as parechoviruses), coxsackievirus A (23 types), and coxsackievirus B (6 types). However, genomic studies have demonstrated that there is significant overlap in the biological characteristics of different serotypes and, more recently, isolated enteroviruses are now named with consecutive numbers (eg, EV68, EV69).

 

The normal site of enterovirus replication is the gastrointestinal tract where the infection is typically subclinical. However, in a proportion of cases, the virus spreads to other organs, causing systemic manifestations, including mild respiratory disease (eg, common cold); conjunctivitis; hand, foot, and mouth disease; aseptic meningitis; myocarditis; and acute flaccid paralysis. Collectively, enteroviruses are the most common cause of upper respiratory tract disease in children. In addition, the enteroviruses are the most common cause of central nervous system (CNS) disease; they account for almost all viruses recovered in culture from spinal fluid. Differentiation of enteroviruses from other viruses and bacteria that cause CNS disease is important for the appropriate medical management of these patients.

 

Traditional cell culture methods require 6 days, on average, for enterovirus detection. In comparison, real-time polymerase chain reaction (PCR) allows same-day detection. Detection of enterovirus nucleic acid by PCR is also the most sensitive diagnostic method for the diagnosis of CNS infection caused by these viruses.

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.

Negative

Interpretation
Provides information to assist in interpretation of the test results

A positive result indicates the presence of enterovirus RNA in the specimen.

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 rule out the possibility of enterovirus infection.

 

This assay may detect virus from a variety of specimen types in asymptomatic individuals. This assay should only be used for patients with a clinical history and symptoms consistent with enterovirus infection and must be interpreted in the context of the clinical picture.

Supportive Data

Accuracy/Diagnostic Sensitivity and Specificity:

Mayo Clinic Laboratories compared the generic detection of enteroviruses from spinal fluid by conventional tube cell culture (MCR-5) and by LightCycler polymerase chain reaction (PCR). Of 715 specimens tested, enteroviruses were detected in 65 (9%) by conventional cell culture and 82 (11%) by LightCycler PCR. Twenty-two of 82 (27%) were exclusively positive by PCR, whereas only 5 of 65 (8%) were exclusively positive by conventional cell cultures.

 

Supplemental Data (Spiking Studies):

To supplement the data above, 30 or more negative specimens of each specimen type (cerebrospinal fluid/sterile body fluid, dermal/ocular/rectal swabs, plasma, and upper and lower respiratory specimens) were spiked with enterovirus culture control at approximately 10 to 50 targets/mcL (the approximate limit of detection). The spiked specimens were run in a blinded manner along with negative (non-spiked) specimens of each specimen type. Of the spiked specimens, 97% to 100% were positive, and 100% of the non-spiked specimens were negative. A total of 489 spiked and non-spiked specimens were tested.

 

Assay Inclusivity:

The assay detected all 64 members of an enterovirus panel, consisting of coxsackieviruses, polio viruses, echoviruses, and other enteroviruses.

 

Analytical Specificity/Limit of Detection:

The lower limit of detection of this assay is approximately 10 to 50 RNA target copies/mcL. This was confirmed in all specimen types accepted for this assay.

 

Specificity:

The assay did not crossreact with a specificity panel containing other RNA-containing viruses (rhinovirus; reovirus; influenza virus, types A and B; respiratory syncytial virus; and parainfluenza virus) and DNA-containing viruses (herpes simplex, Epstein-Barr virus, varicella-zoster virus, and cytomegalovirus).

 

Reportable Range:

This is a qualitative assay and results are reported as either negative or positive for targeted enterovirus RNA.

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. Harvala H, Broberg E, Benschop K, et al: Recommendations for enterovirus diagnostics and characterisation within and beyond Europe. J Clin Virol. 2018 Apr;101:11-17

2. Khetsuriani N, Lamonte-Fowlkes A, Oberst S, et al: Enterovirus surveillance-United States, 1970-2005. MMWR Surveill Summ, 2006 Sep 15;55(8):1-20

3. Foray S, Pailloud F, Thouvenot D, Floret D, Aymard M, Lina B: Evaluation of combining upper respiratory tract swab samples with cerebrospinal fluid examination for the diagnosis of enteroviral meningitis in children. J Med Virol. 1999 Feb;57(2):193-197

4. Furione M, Zavattoni M, Gatti M, Percivalle E, Fioroni N, Gerna G: Rapid detection of enteroviral RNA in cerebrospinal fluid (CSF) from patients with aseptic meningitis by reverse transcription-nested polymerase chain reaction. New Microbiol. 1998 Oct;21(4):343-351

Method Description
Describes how the test is performed and provides a method-specific reference

For this real-time reverse-transcription laboratory-developed polymerase chain reaction (PCR) assay, viral nucleic acid is extracted from specimens using the MagNA Pure automated instrument (Roche Applied Science), followed by amplification and detection on the Roche LightCycler 2.0 instrument. This PCR assay has been optimized to detect a target sequence in the polyprotein region. Primers amplify a 193 base-pair product.

 

Enterovirus genomic RNA is first transcribed to complementary DNA (cDNA) by reverse transcriptase, followed by amplification of the cDNA product. The LightCycler instrument can rapidly (30-40 minutes) detect 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 light of a different wavelength that can be measured with a signal proportional to the amount of specific PCR product. FRET (with subsequent production of a detectable fluorescent signal) only occurs when the probes have specifically annealed to the target sequence of the amplicon.

 

Melting curve analysis is performed following PCR amplification and is the detection phase of the assay, since it offers greater sensitivity than the amplification phase and maintains high specificity.

 

The melting phase of the assay occurs as follows:

Starting at 45 degrees C, which allows the probes to bind to the amplified product, the temperature in the thermal chamber is then slowly raised to 80 degrees C and the fluorescence measured at frequent intervals to determine the point where half of the fluorescence is lost as the probes are denatured (ie, "melt") off of the target. This is called the melting temperature (Tm) of that virus. Analysis of the PCR amplification and probe melting curves is accomplished through the use of LightCycler software.(Bernard PS, Reiser A, Pritham GH: Mutation detection by fluorescent hybridization probe melting curves. In: Meuer S, Wittwer C, Nakagawara K, eds. Rapid Cycle Real-Time PCR Methods and Applications. Springer; 2012:11-20)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Sunday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

Same day/1 to 5 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

1 week

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees :
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

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  • Prospective clients should contact their account representative. For assistance, contact Customer Service.

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 instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US 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 Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

87498

LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
ENTP Enterovirus PCR, P 97151-5
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
56068 Enterovirus PCR, P 97151-5

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports