Test Code DENGC Dengue Virus, Molecular Detection, PCR, Spinal Fluid
Ordering Guidance
The presence of dengue virus nucleic acid in cerebrospinal fluid or serum overlaps with the presence of dengue virus nonstructural protein 1 (NS1) antigen (DNSAG / Dengue Virus NS1 Antigen, Serum). Patients with a history of symptoms for more than 1 week may be negative by molecular tests (ie, real-time polymerase chain reaction) and may require serologic testing (DENVP / Dengue Virus Antibody/Antigen Panel, Serum) to confirm the diagnosis of dengue virus infection.
Specimen Required
Collection Container/Tube:
Preferred: Vial number 2
Acceptable: Any vial number
Submission Container/Tube: Sterile screw cap vial
Specimen Volume: 0.5 mL
Collection Instructions: Do not centrifuge or heat inactivate.
Useful For
Aiding in the diagnosis of central nervous system infection caused by dengue virus
Special Instructions
Method Name
Real-Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)
Reporting Name
Dengue Virus, PCR, CSFSpecimen Type
CSFSpecimen Minimum Volume
0.3 mL
Specimen Stability Information
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
CSF | Refrigerated (preferred) | 7 days | |
Frozen | 7 days |
Reject Due To
Heat-inactivated specimen | Reject |
Clinical Information
Dengue virus (DV) is a globally distributed flavivirus with 4 distinct serotypes (DV-1, -2, -3, -4) primarily transmitted by the Aedes aegypti mosquito, which is found throughout the tropical and subtropical regions of over 100 countries. DV poses a significant worldwide public health threat with approximately 2.5 to 3 billion people residing in DV endemic areas, among whom 100 to 200 million individuals will be infected and approximately 30,000 patients will succumb to the disease annually.
Following dengue infection, the incubation period varies from 3 to 7 days. While some individuals remain asymptomatic, the majority will develop classic dengue fever. Symptomatic patients become acutely febrile and present with severe musculoskeletal pain, headache, retro-orbital pain, and a transient macular rash, most often observed in children. Fever defervescence signals disease resolution in most individuals. However, children and young adults remain at increased risk for progression to dengue hemorrhagic fever and dengue shock syndrome, particularly during repeat infection with a new DV serotype.
Detection of DV nucleic acid in cerebrospinal fluid (CSF) is a marker for central nervous system infection caused by this virus. Importantly, the period of time that the virus can be detected in serum and CSF is brief and, therefore, molecular testing should be performed within the first week following onset of symptoms. After this time, serologic testing is the preferred method for diagnosis of DV infection.
Reference Values
Negative
Reference values apply to all ages.
Interpretation
Positive:
The detection of dengue virus nucleic acid in cerebrospinal fluid (CSF) is consistent with acute-phase infection of the central nervous system.
Dengue virus nucleic acid may be detectable during the first 1 to 7 days following the onset of symptoms.
Negative:
The absence of dengue nucleic acid in CSF is consistent with the lack of acute-phase infection.
Dengue virus nucleic acid may not be detected if the CSF specimen is collected immediately following dengue virus infection (<24-48 hours) and is rarely detectable following 7 days of symptoms.
Cautions
Results should be used in conjunction with clinical presentation and exposure history.
Negative dengue virus (DV) polymerase chain reaction results may occur if the specimen was collected more than 7 days following symptom onset. Serologic testing for the presence of IgM and IgG antibodies to DV is recommended in such cases.
Supportive Data
Assay Inclusivity:
The Altona RealStar Dengue virus reverse transcription polymerase chain reaction (RT-PCR) assay was tested using control strains of each of the 4 dengue serotypes and was able to detect serotypes 1, 2, 3 and 4.
Accuracy:
A commercial panel (SeraCare) of known positive samples for dengue virus serotypes 1, 2, 3, and 4 was tested. Each member of the panel was tested in triplicate, and all replicates were positive by the Altona RealStar Dengue assay.
Thirty analyte-negative CSF samples were spiked (1:10 dilution) with plasma samples collected in South America during an outbreak of dengue virus and determined to be positive for the virus. Of the 30 spiked CSF samples, 29 (97%) were positive by the Altona RealStar Dengue RT-PCR assay.
Limit of Detection:
The limit of detection in CSF was determined to be the following:
Dengue serotype 1: 1.4 genomic targets/mcL (700 genomic targets/mL)
Dengue serotype 2: 2 genomic targets/mcL (1000 genomic targets/mL)
Dengue serotype 3: 1.6 genomic targets/mcL (800 genomic targets/mL)
Dengue serotype 4: 1.3 genomic targets/mcL (650 genomic targets/mL)
Reference Range (Analytical Specificity):
Twenty CSF samples collected for non-infectious diseases testing (eg, chemistry) were analyzed by the Altona RealStar Dengue RT-PCR assay, and all 20 were negative.
A cross-reactivity panel of bacteria (n=12), viruses (n=15), and parasites (n=2) was tested, and all were negative by the Altona Dengue RT-PCR assay.
Method Description
The Altona Real Star DENV is a qualitative, reverse transcription-polymerase chain reaction (RT-PCR) assay targeting the 3' untranslated region polyprotein gene. The assay includes a heterologous amplification system (internal control: IC) to identify possible RT-PCR inhibition and to confirm the integrity of the reagents of the kit. Specimens are run on the LightCycler 480 following nucleic acid extraction using the NucliSENS EasyMag (BioMerieux). Real-time RT-PCR technology utilizes a reverse-transcriptase reaction to convert RNA into complementary DNA, PCR for the amplification of specific target sequences, and target specific probes for the detection of the amplified DNA. The probes are labelled with fluorescent reporter and quencher dyes. Probes specific for DENV RNA are labelled with the fluorophore FAM. The probe specific for the IC is labeled with the fluorophore JOE. Using probes linked to distinguishable dyes enables the parallel detection of DENV specific RNA and the IC in corresponding detector channels of the real-time PCR instrument.(Package insert: RealStar Dengue RT-PCR Kit 2.0. Altona Diagnostics; 01/2017)
Day(s) Performed
Tuesday, Thursday
Report Available
Same day/1 to 5 daysSpecimen Retention Time
7 daysPerforming Laboratory
Mayo Clinic Laboratories in RochesterTest 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 |
---|---|---|
DENGC | Dengue Virus, PCR, CSF | 77958-7 |
Result ID | Test Result Name | Result LOINC Value |
---|---|---|
606371 | Dengue Virus, PCR, CSF | 77958-7 |
Forms
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244)with the specimen.