Developing Sandwhich Elisa Kit for Dengue

Title: Developing Sandwhich ELISA Kit for Dengue

Description:

Dengue is a disease caused by a family of viruses that are transmitted by mosquitoes.The virus is transmitted to humans through the bites of infected female mosquitoes.Dengue fever can vary from mild to severe; the more severe forms include dengue shock syndrome and dengue hemorrhagic fever (DHF). Patients who develop the more serious forms of dengue fever usually need to be hospitalized. Dengue should be suspected when there is a high fever (40°C) accompanied by the following symptoms like severe headache, muscle and joint pains, nausea, vomiting, swollen glands or rashes. Symptoms usually last for 2–7 days after the bite from an infected mosquito. There is no specific treatment or vaccine for dengue fever. Detection of the dengue NS1 antigen during the symptomatic phase of illness represents an important advance in the diagnosis of dengue fever. The acquired immune response for a dengue infection consists of the production of IgM and IgG antibodies primarily directed against the virus envelope proteins. Thus, Sandwhich ELISA method can be used in order to detect the dengue NS1 antigen by collectiong patient’s blood. The sanwhich ELISA measures the amount of antigen between two layers of antibodies.

Principle:

This test is designed for in-vitro qualitative detection of Dengue IgG antibodies in human serum or Plasma and is used as a screening test for testing of collected blood samples suspected for Dengue.The microwells are coated with anti dengue NS1 antibodies with high reactivity for NS1 antigen. The samples are added to the wells followed by addition of enzyme conjugate (monoclonal anti dengue NS1 antibodies linked to Horseradish peroxidase (HRPO)) . Antibodies to Dengue if present in the specimen, will bind to the anti- human IgG antibodies adsorbed onto the surface of the wells. A sandwich complex is formed in the wells. Dengue NS1 from serum sample is sandwiched between the antibody and antibody HRPO conjugate.Unbound conjugate is then washed off with wash buffer. The amount of bound peroxidase is proportional to the concentration of dengue NS1 antigen present in the sample. H2SO4 is added to convert the enzymes into a colored product and can be measured by ELISA reader.