Dengue NS1 Antigen ELISA Kit

Sample

human serum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

Dengue virus NS1 Antigen ELISA Kit for early detection of Dengue virus (DENV), is an ELISA assay system for the detection of NS1 antigen in human serum. It is not intended to screen blood or blood components, and is for investigational use only. Not for use in diagnostic procedures.

Contents of Kit

The Dengue virus NS1 Antigen ELISA Kit contains sufficient reagents for one plate of 96 wells (12×8 strips) each.
Warning: Do not use any reagents where damage to the packaging has occurred.
Dengue virus NS1 Antigen ELISA Kit supplied materials:
1. Coated Microtiter Strips for DENV Detect NS1 ELISA:
ELISA strip holder in a ziplock foil pouch with desiccant, containing 96 polystyrene microtiter wells coated with anti-NS1 antibody in each well. Stable at 2-8°C until the expiration date.
2. Negative Control for DENV Detect NS1 ELISA:
One vial, 300μL containing heat-inactivated negative control serum. The negative control will aid in verifying the validity of the kit. Stable at 2-8°C until the expiration date of the kit.
3. Positive Control for DENV Detect NS1 ELISA:
One vial, 300μL containing recombinant NS1 in a phosphatebased buffer with 0.05% Proclin-300. The positive control will aid in verifying the validity of the kit. Stable at 2-8°C until the expiration date.
4. Cut-off Control for DENV Detect NS1 ELISA:
One vial, 300μL containing recombinant NS1 in a phosphate-based buffer with 0.05% Proclin-300. The Cut-off Control will aid in determining the cut-off value for the ELISA. Stable at 2-8°C until the expiration date.
5. Sample Dilution Buffer for DENV Detect NS1 ELISA:
One bottle, 15mL, containing the secondary antibody in a Tris-based buffer with 0.02%-0.05% Proclin-300. Stable at 2-8°C until the expiration date.
6. 100× Conjugate for DENV Detect NS1 ELISA:
One vial, 150μL, containing horseradish peroxidase-labeled polyclonal antibody in a Tris-based buffer with 0.03% - 0.05% Proclin-300. Stable at 2-8°C until the expiration date.
7. Conjugate Diluent for DENV Detect NS1 ELISA:
One bottle, 12mL. This contains the diluent solution for the 100× Conjugate in a Tris-based buffer with 0.01% Thimerosal as a preservative. The 100× conjugate is diluted directly into this solution. After diluting 100× Conjugate into this solution, the now ready-to-use conjugate may be stored up to 2 weeks at 2-8°C before it should be discarded. Stable at 2-8°C until the expiration date.
8. 10× Wash Buffer:
One bottle, 120 mL. 10× concentrated phosphate buffered saline with Tween 20 (pH 6.8-7.0). Stable at 2-8°C until the expiration date.
9. Liquid TMB Substrate:
One bottle, 12mL, ready to use. Contains 3, 3', 5, 5'-tetramethylbenzidine (TMB) and hydrogen peroxide in a citric acid-citrate buffer (pH 3.3-3.8). Stable at 2-8°C until the expiration date. Note: The substrate should always be stored in the light protected bottle provided.
10. Stop Solution:
One bottle, 6mL, read to use 1N Sulfuric Acid. Used to stop the reaction. Stable at 2-8°C until the expiration date.
Warning: Strong acid. Wear protective gloves, mask and safety glasses. Dispose all materials according to all applicable safety rules and regulations.

Storage

Stable at 2-8°C until the expiration date.

Precision

The precision study of the Dengue virus NS1 Antigen ELISA Kit was performed at 3 different sites by 2 different individuals (6 operators total) on 5 separate days. All samples (including controls) were run in triplicate. Four serum specimens (Panels 1-4) using clinical specimens diluted in an analyte-negative matrix, plus a positive control, a negative control and cut-off control, were used. The four serum specimens included a positive specimen, two weakly positive specimens and a negative specimen. The ISR total precision %CV varied from 3.7-11.9%, depending on the sample. The results are shown in the following table.

Sensitivity

The analytical reactivity was performed to determine the limits of detection (LODs) of Dengue virus NS1 Antigen ELISA Kit when tested with native NS1 from viral culture supernatants representing all four serotypes of dengue virus. Dilutions were tested in replicates of 20, and the lowest concentrations at which ≥95% of replicates tested positive were considered the LOD for each serotype. The LOD of Dengue virus NS1 Antigen ELISA Kit ranges from 82-611 pfu/mL, depending upon DENV serotype, as shown in Table below.

General Description

Dengue Fever (DF) is an acute viral disease of man, which is transmitted by Aedes aegypti mosquitoes. DF is characterized clinically by biphasic fever, rash and hematopoietic depression, and by constitutional symptoms such as malaise, arthralgia, myalgia and headache. Infrequently, more severe disease is seen, manifested by hemorrhagic fever which may progress to lethal shock. It is endemic in the tropics and subtropics, worldwide, where an estimated 100,000,000 cases occur annually. It has been estimated that about 50 to 100 million cases of DF occur every year with about 250,000 to 500,000 cases of Dengue Hemorrhagic Fever (DHF). During 2002, more than 30 Latin American countries reported over 10,000,000 DF cases with large number of DHF cases. This has been followed by extensive epidemics of DHF in several parts of India during 2003 through 2005. In the Americas, the reported incidence has more than tripled from 1996 to 2002. The incidence of Dengue outbreak has been reported in Hawaii, and in Laredo, Texas. Dengue NS1 (non-structural) protein is a secreted protein and is believed to play a role in viral RNA replication. NS1 is strongly immunogenic, eliciting antibodies with complement fixing activity. NS1 antigen can be detected in circulating blood during acute Dengue infection. The Dengue virus NS1 Antigen ELISA Kit can detect NS1 antigen in serum samples within 1 to 2 days following infection.