Test ID: LHSV
Herpes Simplex Virus (HSV), Molecular Detection, PCR
Secondary ID 
A test code used for billing and in test definitions created prior to November 2011
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
Useful For Suggests clinical disorders or settings where the test may be helpful
Aiding in the rapid diagnosis of herpes simplex virus (HSV) infections, including qualitative detection of HSV DNA in cerebrospinal fluid and other (non-blood) clinical specimens
Qualitative detection of HSV DNA
Method Name A short description of the method used to perform the test
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
Aliases Lists additional common names for a test, as an aid in searching
HSV (Herpes Simplex Virus) by PCR (Polymerase Chain Reaction)
HSV Detection by Real-Time PCR
HSV PCR
LightCycler HSV
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.
Specimen source is required.
Submit only 1 of the following specimens:
Preferred:
Specimen Type: Body or ocular fluid
Container/Tube: Sterile container
Specimen Volume: 0.5 mL
Collection Instructions: The high sensitivity of amplification by PCR requires the specimen to be processed in an environment in which contamination of the specimen by HSV DNA is not likely.
Specimen Stability Information: Refrigerated (preferred)/Frozen
Specimen Type: Genital
Sources: Cervix, rectum, urethra, vagina, other genital site
Container/Tube: BBL CultureSwab (Supply T092)
Specimen Volume: Swab
Collection Instructions: Place swab back into swab cylinder.
Specimen Stability Information: Refrigerated (preferred)/Frozen
Specimen Type: Respiratory
Sources: Bronchial washing, bronchoalveolar lavage, nasopharyngeal aspirate or washing, sputum, tracheal aspirate
Container/Tube: Sterile container
Specimen Volume: 1.5 mL
Specimen Stability Information: Refrigerated (preferred)/Frozen
Specimen Type: Spinal fluid
Container/Tube: Sterile vial
Specimen Volume: 0.5 mL
Collection Instructions: Do not centrifuge.
Specimen Stability Information: Refrigerated (preferred)/Frozen
Additional Information: The high sensitivity of amplification by PCR requires the specimen to be processed in an environment in which contamination of the specimen by HSV DNA is not likely.
Specimen Type: Swab
Sources:
Preferred: Dermal, eye, throat
Acceptable: Anal/rectal, urethral
Container/Tube: BBL CultureSwab (Supply T092)
Specimen Volume: Swab
Collection Instructions: Place swab back into swab cylinder.
Specimen Stability Information: Refrigerated (preferred)/Frozen
Specimen Type: Tissue
Sources: Brain, colon, kidney, liver, lung, etc.
Container/Tube: Sterile container containing 1 to 2 mL of sterile saline or multi-microbe medium (M5) (Supply T484)
Specimen Volume: Entire collection
Collection Instructions: Collect a fresh tissue specimen.
Acceptable:
Specimen Type: Amniotic fluid
Container/Tube: Amniotic fluid container
Specimen Volume: 0.5 mL
Specimen Type: Urine (<1 month old infant)
Container/Tube: Sterile container
Specimen Volume: 0.5 mL
Specimen Type: Whole blood (<1 month old infant)
Container/Tube: EDTA
Specimen Volume: 0.5 mL
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.
Reject Due To Identifies specimen types and conditions that may cause the specimen to be rejected
| Hemolysis | NA |
| Lipemia | NA |
| Icterus | NA |
| Other | Calcium alginate-tipped swab, wood swab, or transport swab containing gel; formalin-fixed and/or paraffin-embedded tissues |
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
| Specimen Type | Temperature | Time |
|---|---|---|
| Varies | Varies | 7 days |
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Herpes simplex virus (HSV) causes various clinical syndromes. Anatomic sites infected include skin, lips, oral cavity, eyes, genital tract, and central nervous system (CNS). Systemic involvement may also occur.
Fresh brain tissue is the definitive specimen for detection of HSV from patients with CNS disease. However, because brain biopsy is an invasive procedure, it is infrequently performed for laboratory diagnosis. Similarly, it is difficult to recover HSV from cerebrospinal fluid (CSF) specimens in culture systems, and the serologic diagnosis of HSV CNS disease has not been informative during early onset disease. HSV PCR detection from CSF is a sensitive and specific alternative for detection of disease involving the CNS, as well as oral, genital, ocular, and other sites.
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.
Not applicable
Interpretation Provides information to assist in interpretation of the test results
This is a qualitative assay; results are reported either as negative or positive for herpes simplex virus (HSV) type 1 or HSV type 2, or HSV indeterminate.
Detection of HSV DNA in clinical specimens supports the clinical diagnosis of infection due to the virus. The lower limit of detection of LightCycler PCR is 10 copies DNA target per microliter. HSV DNA is not detected in cerebrospinal fluid from patients without central nervous system disease caused by this virus.
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 eliminate the possibility of herpes simplex virus (HSV) infection. HSV DNA may not be detectable in the early acute stages of the central nervous system disease. In addition, in some cases, after initial detection (positive result), HSV DNA may only be present in cerebrospinal fluid (CSF) for 3 to 4 weeks after initial presentation of symptoms. DNA levels may fall to undetectable with time.
Although the reference range is typically "negative" for this assay, this assay may detect viral shedding in asymptomatic individuals. This may be especially relevant when dermal or genital sites are tested, since intermittent shedding without noticeable lesions has been described.(1) CSF DNA is not expected to contain detectable HSV DNA in patients without related disease. This assay is only to be used for patients with a clinical history and symptoms consistent with HSV infection, and must be interpreted in the context of the clinical picture. This test should not be used to screen asymptomatic patients.
Supportive Data
Accuracy/Diagnostic Sensitivity and Specificity:
Of 200 specimens processed by both shell vial assay and LightCycler, herpes simplex virus (HSV) was detected in 88 specimens (44%). All 88 positive specimens were detected by LightCycler compared with 69 by the shell vial assay. The 19 discrepant results (LightCycler positive, shell vial assay negative) were resolved as true positive results by using a PCR assay directed to another gene target (thymidine kinase) of the virus.
Of 100 CSF specimens tested by both conventional PCR and LightCycler, HSV was detected in 38 specimens by both techniques. Four specimens were detected only by conventional PCR; however 6 specimens were detected exclusively by LightCycler. Fifty-two specimens were found to be negative for HSV by both techniques.
Supplemental Data (Spiking Studies):
To supplement the above data, approximately 30 negative specimens each of various types were spiked with HSV 1 and HSV 2 plasmid control at the limit of detection (10 copies DNA target/microliter). The spiked specimens were run in a blinded fashion along with approximately 30 negative (non-spiked) specimens each of various specimen types; 92% 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 DNA target copies per microliter. This was established in anogenital swabs and confirmed in each specimen type accepted for this assay.
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:
Inter- and intra-assay precision was 100% and 100%, respectively.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Schiffer JT, Corye L: New concepts in understanding genital herpes. Curr Infect Dis Rep Nov 2009;11(6):457-464
2. Espy MJ, Uhl JR, Svien KA: Laboratory diagnosis of herpes simplex virus infections in the clinical laboratory by LightCycler PCR. J Clin Microbiol 2000;38(2):795-799
3. Espy MJ, Ross TK, Teo R: Evaluation of LightCycler PCR for implementation of laboratory diagnosis of herpes simplex virus infections. J Clin Microbiol 2000;38(8):3116-3118
4. Sauerbrei A, Eichhorn U, Hottenrott G, Wutzler P: Virological diagnosis of herpes simplex encephalitis. J Clin Virol 2000;17(1):31-36
5. Mitchell PS, Espy MJ, Smith TF, et al: Laboratory diagnosis of central nervous system infections with herpes simplex virus by PCR performed with cerebrospinal fluid specimens. J Clin Microbiol 1997;35(11):2873-2877
6. Yi-Wei T, Mitchell PS, Espy MJ, et al: Molecular diagnosis of herpes simplex virus infections in the central nervous system. J Clin Microbiol 1999;37(7):2127-2136
Method Description Describes how the test is performed and provides a method-specific reference
Viral nucleic acid is extracted by the MagNA Pure automated instrument (Roche Applied Science) from genital, dermal, tissue, or cerebrospinal fluid specimens. Primers directed to the DNA polymerase of herpes simplex virus produce a 215 base pair amplicon. The LightCycler instrument (Roche Applied Science), 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 through stringent air-controlled temperature cycling and 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. LightCycler hybridization probes are designed for HSV-type 2 and sequence differences between HSV-type 2- and HSV-type 1 are detected by melting curve analysis. 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. Sequence differences between the PCR amplification and probe melting curves are accomplished through the use of LightCycler software. (Espy MJ, Uhl JR, Svien KA: Laboratory diagnosis of herpes simplex virus infections in the clinical laboratory by LightCycler PCR. J Clin Microbiol 2000;38[2]:795-799)
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 Saturday; 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.
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.
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.
87529
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 |
|---|---|---|
| SS001 | Specimen Source | 31208-2 |
| 80575 | Result | 5013-8 |
| MSI27 | Special Information | 48767-8 |
| BS024 | Report Status | N/A |
External
link
PDF
document
Excel
document
Word
document