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Test Code LABVZPCR Varicella-Zoster Virus, Molecular Detection, PCR, Varies

Additional Codes

Test Name in EPIC

EPIC Test Code

Mnemonic

Mayo Test ID

VARICELLA-ZOSTER BY PCR, VARIES

 LABVZPCR VZVPV VZVPV

 


Necessary Information


1. Specimen source is required.

2. Source information must include main anatomical site of collection.



Specimen Required


Submit only 1 of the following specimens:

 

Supplies: Sarstedt Aliquot Tube, 5 mL (T914)

Specimen Type: Body fluid

Sources: Spinal, pleural, peritoneal, ascites, pericardial, amniotic, or ocular

Container/Tube: Sterile container

Specimen Volume: 0.5 mL

Collection Instructions: Do not centrifuge.

 

Specimen Type: Swab

Sources: Miscellaneous; dermal, eye, nasal, throat, or genital

Supplies:

-Culturette (BBL Culture Swab) (T092)

-BD E-Swab (T853)

-M4-RT (T605)

Container/Tube: Multimicrobe media (M4-RT, M4, M5, Bartels, or Jiangsu) and E-Swab or Culturette

Collection Instructions: Place swab back into multimicrobe media.  

 

Specimen Type: Respiratory

Sources: Bronchial washing, bronchoalveolar lavage, nasopharyngeal aspirate or washing, sputum, or tracheal aspirate

Container/Tube: Sterile container

Specimen Volume: 1.5 mL

 

Specimen Type: Fresh tissue

Supplies: M4-RT (T605)

Container/Tube:

Preferred: Multimicrobe media (M4-RT, M4, M5, Bartels, or Jiangsu)

Acceptable: Sterile container with 1 to 2 mL of sterile saline

Specimen Volume: Entire collection

Collection Instructions: Submit only fresh tissue. Fixed tissue is not acceptable.


Useful For

Rapid (qualitative) detection of varicella-zoster virus DNA in clinical specimens for laboratory diagnosis of disease due to this virus

 

This test should not be used to screen asymptomatic patients.

Method Name

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

Reporting Name

Varicella-Zoster Virus, PCR, Varies

Specimen Type

Varies

Specimen Minimum Volume

Ocular Fluid and Spinal Fluid: 0.3 mL
Body Fluid (pleural, peritoneal, ascites, and pericardial): See Specimen Required
Respiratory Specimens: 1 mL
Tissue: 2 × 2 mm biopsy

Specimen Stability Information

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

Reject Due To

Calcium alginate-tipped swab
Wood swab
Transport swab containing gel
Formalin-fixed and/or paraffin-embedded tissues
Heat-inactivated specimen Dry/flocked E-Swab		 Reject

Clinical Information

Varicella-zoster virus (VZV) causes both varicella (chickenpox) and herpes zoster (shingles). VZV produces a generalized vesicular rash on the dermis (chickenpox) in normal children, usually before 10 years of age. After primary infection with VZV, the virus persists in latent form and may emerge clinically (usually in adults 50 years of age and older) to cause a unilateral vesicular eruption, generally in a dermatomal distribution (shingles).

Reference Values

Negative

 

Reference values apply to all ages.

Interpretation

Detection of varicella-zoster virus (VZV) DNA in clinical specimens supports the clinical diagnosis of infection due to this virus.

 

VZV DNA is not detected in cerebrospinal fluid from patients without central nervous system disease caused by this virus.

 

This LightCycler polymerase chain reaction assay does not yield positive results with other herpesvirus gene targets (herpes simplex virus, cytomegalovirus, Epstein-Barr virus).

Cautions

A negative result does not exclude the possibility of varicella-zoster virus (VZV) infection.

 

The reference range is typically "negative" for this assay. This assay is only to be used for patients with a clinical history and symptoms consistent with VZV infection and must be interpreted in the context of the clinical picture.

Supportive Data

The following validation data supports the use of this assay for clinical testing.

 

Accuracy/Diagnostic Sensitivity and Specificity:

LightCycler polymerase chain reaction (PCR) (primers, directed to varicella-zoster virus [VZV], gene 29) was compared with shell vial cell cultures for the detection of VZV from 253 dermal specimens. Twenty-three specimens (9.1%) were positive for VZV by LightCycler PCR and the shell vial cell culture assay. An additional 21 specimens exclusively yielded VZV DNA. These discrepant specimens were resolved as true-positive results by confirmation of results by PCR using primers directed to another gene of VZV. Importantly, there were no instances in which VZV was recovered by the shell vial assay and not detected by LightCycler PCR (specificity, 100%). Of 100 cerebrospinal fluid specimens tested by both conventional PCR and LightCycler PCR, VZV DNA was detected in 49 specimens by both methods; 1 specimen was positive only by the conventional PCR assay. Fifty specimens were found to be negative for VZV DNA by both techniques.

 

Supplemental Data (Spiking Studies):

To supplement the above data, 30 negative specimens each various specimen type were spiked with VZV plasmid at the limit of detection (10-20 targets/microliter). The spiked specimens were run in a blinded fashion along with approximately 30 negative (non-spiked) specimens each of various specimen types; 90% to 100% of the spiked specimens were positive and 100% of the non-spiked specimens were negative.

 

Analytical Sensitivity/Limit of Detection:

The limit of detection of this assay is 10 to 20 DNA target copies per microliter in specimen matrix.

 

Analytical Specificity:

No PCR signal was obtained from extracts of 27 bacterial, viral, and fungal isolates that could be found as normal flora in sites normally tested for this organism or that could cause similar symptoms.

 

Precision:

Interassay precision was 100%, and intraassay precision was 97%.

 

Reportable Range:

This test is a qualitative assay, and results are reported as negative or positive for targeted VZV DNA.

Method Description

Viral nucleic acid is extracted by the MagNA Pure automated instrument (Roche Applied Science) from clinical specimens. Primers directed to target DNA (ss DNA binding proteins: gene 29) produce a 202-base pair amplicon. The LightCycler instrument amplifies and monitors by fluorescence the development of target nucleic acid sequences after the annealing step during PCR cycling. This is an automated PCR system that can rapidly detect (30-40 minutes) amplicon development though stringent air-controlled temperature cycling in capillary cuvettes. The detection of amplified products is based on the fluorescence resonance energy transfer 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 probe melting curves is accomplished through the use of LightCycler software.(Dhiman N, Wright PA, Espy MJ, Schneider SK, Smith TF, Pritt BS. Concurrent detection of herpes simplex and varicella-zoster viruses by polymerase chain reaction from the same anatomic location. Diagn Microbiol Infect Dis. 2011;70(4):538-540. doi:10.1016/j.diagmicrobio.2011.03.014; Espy MJ, Teo R, Ross TK, Scien KA, Wold AD, Smith TF. Diagnosis of varicella-zoster virus infections in the clinical laboratory by LightCycler PCR. J Clin Microbiol. 2000;38[9]:3187-3189)

Day(s) Performed

Monday through Sunday

Report Available

Same day/1 to 4 days

Specimen Retention Time

1 week

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Test Classification

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

87798

LOINC Code Information

Test ID Test Order Name Order LOINC Value
VZVPV Varicella-Zoster Virus, PCR, Varies 94584-0

 

Result ID Test Result Name Result LOINC Value
VZVS Specimen Source 31208-2
618332 Varicella-Zoster Virus PCR 94584-0

Forms

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