Dengue Virus IgM Antibody ELISA Assay Kit:
For Research Use Only
Size: 1 x 96 wells
Sensitivity: diagn. >90%
Dynamic Range: Cut-off
Incubation Time: 2 hour
Sample Type: Serum, citrate Plasma
Sample Size: 10 µl
The Eagle Biosciences Dengue Virus IgM Antibody ELISA assay kit is intended for the quantification of Dengue Virus IgM in Human serum and plasma. The Eagle Biosciences Dengue Virus IgM Antibody ELISA assay kit is for research use only and should not be used for diagnostic procedures.
Dengue virus is a single-stranded RNA virus of about 50 nm in diameter belonging to the genus Flavivirus. Dengue and dengue hemorrhagic fever are caused by one of four closely related, but antigenically distinct, virus serotypes (DEN-1, DEN-2, DEN-3, and DEN-4). Infection with one of these serotypes does not provide cross protective immunity, so persons living in a dengue-endemic area can have four dengue infections during their lifetimes. The viruses are transmitted by Aedes aegypti, a domestic, day-biting mosquito that prefers to feed on humans. Infection with dengue viruses produces a spectrum of clinical illness ranging from a nonspecific viral syndrome to severe and fatal hemorrhagic disease. It is primarily a disease of the tropics; its global distribution is comparable to that of malaria, and an estimated 2.5 billion people live in areas at risk for epidemic transmission. – Globally, there are an estimated 50 to 100 million cases of dengue fever and several hundred thousand cases of dengue hemorrhagic fever.
• The case-fatality rate of DHF in most countries is about 5%; most fatal cases are among children and young adults.
• Important risk factors for DHF include the strain and serotype of the infecting virus, as well as the age, immune status, and
• Genetic predisposition of the patient.
• Risk groups: residents of or visitors to tropical urban areas
The presence of virus resp. infection may be identified by
• Serology: Detection of antibodies by ELISA Infection produces lifelong immunity, but the antigenically distinct serotypes do not provide cross-protective immunity, so a person can theoretically experience four dengue infections; a dengue vaccine is not available.
This assay employs the quantitative enzyme immunoassay technique. A specific Dengue Virus antigen Type 2 has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Dengue virus antibody present is bound by the immobilized antigen. After washing away any unbound substances, an HRP-conjugated antibody specific for human IgG is added to each well and incubate. Following the washing of any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of antigen-antibody binding in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ±2nm.
1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
2. Add 100 μl of controls and diluted samples (1:100) into wells. Leave well A1 for substrate blank. Incubate for 1h at 37°C.
3. Aspirate each well and wash, repeating the process 4 times for a total 5 washes. Wash by filling each well with 1× Wash Buffer (300 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
4. Add 100 μl HRP-conjugated antibody (ready-to-use) into each well except for the substrate blank well. Cover wells and incubate for 30 minutes at RT.
5. Aspirate each well and wash as step 3.
6. Add 100 μl of TMB Reagent to each well. Incubate for 15 minutes at room temperature in the dark.
7. Add 100 μl of Stop Solution to each well. The color of the solution should change from blue to yellow.
8. Read the OD with a microplate reader at 450nm immediately.
Intra-assay and Inter-assay precision
The CV value of intra-assay precision is 3.1% and inter-assay precision is 4.8%.