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Test Code DENVP Dengue Virus Antibody/Antigen Panel, Serum

Useful For

Aids in the diagnosis of dengue virus infection

Profile Information

Test ID Reporting Name Available Separately Always Performed
DENG Dengue Virus Ab, IgG, S No Yes
DENM Dengue Virus Ab, IgM, S No Yes
DENS1 Dengue NS1 Ag, S No Yes
INT69 Dengue Interpretation No Yes

Reporting Name

Dengue Virus Ab/Ag Panel, S

Specimen Type

Serum


Specimen Required


Container/Tube:

Preferred: Serum gel

Acceptable: Red top

Specimen Volume: 1 mL


Specimen Minimum Volume

0.8 mL (DENS1: 0.4 mL and DENG, DENM, INT69: 0.4 mL)

Specimen Stability Information

Specimen Type Temperature Time
Serum Refrigerated (preferred) 14 days
  Frozen  14 days

Reject Due To

Hemolysis

Mild OK; Gross reject

Lipemia

Mild OK; Gross reject

Icterus

Mild OK; Gross reject

Other

NA

Clinical Information

Dengue virus (DV) is a globally distributed flavivirus with 4 distinct serotypes (DV-1, -2, -3, -4) and is 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 and while some infections remain asymptomatic, the majority of individuals 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 dengue-specific IgM/IgG-class antibodies remains the most commonly utilized diagnostic method. Seroconversion occurs approximately 3 to 7 days following exposure and therefore testing of acute and convalescent sera may be necessary to make the diagnosis. Detection of the DV nonstructural protein 1 (NS1) has emerged as an alternative biomarker to both serologic- and molecular-based techniques for diagnosis of acute DV infection. NS1 antigenemia is detectable within 24 hours and up to 9 days following symptoms onset. This overlaps with the DV viremic phase and NS1 is often detectable prior to IgM seroconversion. Concurrent evaluation (as performed in this profile) for the NS1 antigen alongside testing for IgM- and IgG-class antibodies to DV provides optimal diagnostic potential for both early and late dengue disease.

Reference Values

IgG: negative

IgM: negative

NS1: negative

Reference values apply to all ages.

Interpretation

IgG:

The presence of IgG-class antibodies to dengue virus (DV) is consistent with exposure to this virus sometime in the past. By 3 weeks following exposure, nearly all immunocompetent individuals should have developed IgG antibodies to DV.

 

IgM:

The presence of IgM-class antibodies to DV is consistent with acute-phase infection.

 

IgM antibodies become detectable 3 to 7 days following infection and may remain detectable for up to 6 months or longer following disease resolution.

 

The absence of IgM-class antibodies to DV is consistent with lack of infection. However, specimens drawn too soon following exposure may be negative for IgM antibodies to DV. If DV remains suspected, a second specimen, drawn approximately 10 to 12 days following exposure should be tested.

 

Nonstructural protein 1 (NS1):

The presence of dengue NS1 antigen is consistent with acute-phase infection with dengue virus.

 

The NS1 antigen is typically detectable within 1 to 2 days following infection and up to 9 days following symptom onset.

 

NS1 antigen may also be detectable during secondary dengue virus infection, but for a shorter duration of time (1-4 days following symptom onset).

 

The absence of dengue NS1 antigen is consistent with the lack of acute-phase infection.

 

The NS1 antigen may be negative is samples collected immediately following dengue virus infection (<24-48 hours) and is rarely detectable following 9 to 10 days of symptoms.

Cautions

Test results should be used in conjunction with clinical evaluation, including exposure history and clinical presentation.

 

False-positive results, particularly with the dengue virus (DV) IgG enzyme-linked immunosorbent assay (ELISA), may occur in persons infected with other flaviviruses, including West Nile virus and St. Louis encephalitis virus. Obtaining a detailed exposure history and further laboratory testing may be necessary to determine the infecting virus.

 

Positive test results may not be valid in persons who have received blood transfusions or other blood products within the last several months.

 

The significance of a negative result in an immunosuppressed patient is unclear.

 

Results should be used in conjunction with clinical presentation and exposure history.

 

Though uncommon, false-positive nonstructural protein 1 (NS1) results may occur in individuals with active infection due to other flaviviruses, including West Nile virus and yellow fever virus.

 

Negative NS1 antigen 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

A total of 200 prospective serum samples submitted for dengue virus (DV) IgM and IgG testing by the Focus Diagnostics DV IgM and IgG EIAs were also tested by the InBios IgM and IgG DV assays. The results were compared and the data summarized in Tables 1 and 2.

 

Table 1. Comparison of the InBios and Focus Diagnostics DV IgM EIAs

 

Focus Diagnostics DV IgM EIA

 

 

Positive

Negative

InBios

DV IgM EIA

Positive

14

0

Negative

1

184

Equivocal

1

0

 

Sensitivity: 87.5% (14/16); 95% Confidence Intervals (C.I.) 62.7%-97.7%

Specificity: 100% (184/184); 95% C.I. 97.5%-100%

Agreement: 99% (198/200); 95% C.I. 96.1%-99.9%

 

Table 2. Comparison of the InBios and Focus Diagnostics DV IgG EIAs

 

Focus Diagnostics DV IgG EIA

 

 

Positive

Negative

InBios

DV IgG EIA

Positive

34

0

Negative

0

164

Equivocal

2

0

 

Sensitivity: 94.4% (34/36); 95% C.I. 80.9%-99.4%

Specificity: 100% (164/164); 95% C.I. 97.2%-100%

Agreement: 99% (198/200); 95% C.I. 96.1%-99.9%

 

An additional 42 serum samples positive for IgG-class antibodies to West Nile virus (n=24), St. Louis encephalitis virus (n=9) and California (LaCrosse) virus (n=9) were also tested by the InBios DV IgG EIA and the data are summarized below in Table 3.

 

Table 3. Cross-reactivity of the InBios DV IgG EIA with antibodies to West Nile virus, St. Louis encephalitis virus and California (LaCrosse) virus

 

 

West Nile Virus IgG Positive

St. Louis Encephalitis Virus Positive

California (LaCrosse) Virus Positive

InBios

DV IgG EIA

Positive

18

7

1

Negative

2

0

8

Equivocal

4

2

0

 

Note that the InBios DV IgG EIA shows significant cross-reactivity with antibodies to West Nile virus and St. Louis encephalitis virus.

 

The presence of nonstructural protein 1 (NS1) antigen overlaps with the DV viremic phase for the first 4 to 5 days following infection and therefore, the performance characteristics of the InBios DV NS1 EIA were compared to the Focus Diagnostics DV real-time PCR (RT-PCR), which detects RNA from all 4 DV serotypes. Seventy-seven serum samples previously evaluated by the Focus Diagnostics RT-PCR assay were also tested by the InBios DV NS1 EIA and the results are compared in Table 4 below. Discordant samples were also tested by the Platelia NS1 Ag EIA (BioRad Laboratories, Marnes-la-Coquette, France).

 

Table 4. Comparison of the InBios NS1 EIA to RT-PCR for DV Detection

 

Focus Diagnostics DV RT-PCR

 

 

Positive

Negative

InBios

DV NS1 EIA

Positive

24

7(b)

Negative

1(a)

43

Equivocal

0

2(c)

 

a. This sample was negative by the Platelia NS1 EIA

b. Five samples were also positive by the Platelia NS1 EIA

c. One sample was negative and 1 sample was indeterminate by the Platelia NS1 EIA

 

Sensitivity: 96% (24/25); 95% Confidence Intervals (C.I.): 79.1%-100%

Specificity: 82.7% (43/52); 95% C.I.: 70.1%-90.9%

Overall Agreement: 87.1% (67/77); 95% C.I.: 77.6%-93%

Method Description

DENV Detect IgM Capture enzyme-linked immunosorbent assay (ELISA):

Samples and controls are diluted in Sample Dilution Buffer and incubated in microtiter wells coated with antihuman IgM antibodies. This incubation is followed by incubation with dengue-derived recombinant antigens (DENRA) and normal cell antigen (NCA) separately. After incubation and washing, the wells are treated with a DEN-specific monoclonal antibody labeled with horseradish peroxidase (HRP). After a second incubation and washing step, the wells are incubated with tetramethylbenzidine (TMB) substrate.Acid stop is added and absorbance at 450 nm is read. Ratio of absorbencies of the DENRA and the control antigen wells determine whether the result is positive or negative.(Package insert: InBiOS DENV Detect IgM CAPTURE ELISA. version 900106-03 8/16/2011.InBios International, Inc, Seattle WA)

 

DENV Detect IgG ELISA:

Controls and diluted samples are incubated in microtiter wells coated with monoclonal antibody bound to DENRA. After incubation and washing, wells are treated with IgG antibody labeled with HRP. After a second incubation and washing, wells are incubated with TMB substrate. Acid stop is added and absorbance at 450 nm is measured. Ratio of the absorbencies of the DENRA and the control wells determines whether a result is positive or negative.(Package insert: InBiOS DENV Detect IgG ELISA version 900089.1. InBios International, Inc, Seattle WA)

 

The InBios nonstructural protein 1 (NS1) ELISA is a 2-step sandwich-format colorimetric immunoassay for qualitative detection of NS1 antigen in serum. Testing is performed according to manufacturer's instructions on the Triturus automated EIA analyzer (Grifols, Miami, FL). Diluted patient samples and controls incubated in wells coated with purified capture antibody, specific for the dengue NS1 antigen. Following incubation, wells are washed, incubated with a horseradish peroxidase-conjugated polyclonal antibody specific to NS1 antigen and reincubated. Wells are subsequently washed and 3,3',5,'-tetramethlbenzidine substrate is added and incubated at room temperature (RT) in the dark. Stop solution is added next and the optical density (OD) of the reaction is measured at 450/620 nm. The immune status ratio (ISR) for each sample is calculated from the ratio of the OD obtained with the test sample divided by the OD from the calculated cutoff value (determined by the cutoff control sample).(Package insert: InBios DENV Detect NS1 ELISA, Seattle, WA)

Day(s) and Time(s) Performed

Tuesday; 9 a.m.

Analytic Time

Same day/1 day

Performing Laboratory

Mayo Medical Laboratories in Rochester

CPT Code Information

IgG: 86790

IgM: 86790

NS1: 86790

LOINC Code Information

Test ID Test Order Name Order LOINC Value
DENVP Dengue Virus Ab/Ag Panel, S In Process

 

Result ID Test Result Name Result LOINC Value
INT69 Dengue Interpretation 69048-7
DENG Dengue Virus Ab, IgG, S 29661-6
DENM Dengue Virus Ab, IgM, S 29663-2
DENS1 Dengue NS1 Ag, S 75377-2

Method Name

Enzyme-Linked Immunosorbent Assay (ELISA)

Specimen Retention Time

14 days

Test Classification

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.