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Product Name: Dengue Virus IgM - ELISA
# of Samples: 1 x 96 Assays
Intended Use: The Dengue virus IgM-ELISA is intended for the qualitative determination of IgM class antibodies against Dengue virus in human serum or plasma (citrate).
Introduction: 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
Principles of the assay: The qualitative immunoenzymatic determination of IgM-class antibodies against Dengue Virus is based on the ELISA (Enzyme-linked Immunosorbent Assay) technique.
Microtiter strip wells are precoated with Dengue Virus antigens type 2 to bind corresponding antibodies of the specimen. After washing the wells to remove all unbound sample material horseradish peroxidase (HRP) labelled anti-human IgM conjugate is added. This conjugate binds to the captured Dengue Virus-specific antibodies. The immune complex formed by the bound conjugate is visualized by adding Tetramethylbenzidine (TMB) substrate which gives a blue reaction product. The intensity of this product is proportional to the amount of Dengue Virus-specific IgM antibodies in the specimen. Sulphuric acid is added to stop the reaction. This produces a yellow endpoint colour. Absorbance at 450 nm is read using an ELISA microwell plate reader.
Storage and Stability: The reagents are stable up to the expiry date stated on the label when stored at 2...8 °C.
Limitations of the Test: The Flaviviridae family includes the serotypes of Dengue virus as well as the yellow fever and Japanese encephalitis viruses. There is a cross reactivity among flaviviruses, due to the presence of common antigenic determinants.
Bacterial contamination or repeated freeze-thaw cycles of the specimen may affect the absorbance values. Diagnosis of an infectious disease should not be established on the basis of a single test result. A precise diagnosis should take into consideration clinical history, symptomatology as well as serological data. It is recommended to test a reactive sample with a second serological method.
In immunocompromised patients and newborns serological data only have restricted value.
References Hayes E.B.,Gubler D.J.,Dengue and dengue hemorrhagic fever,Pediatric Inf.Dis.J.11:311-317,1992CDC: Imported Dengue-United States 1993-1994,J.Am.Med.Assn.274:113,1995 Rigau-Perez J.G.,Gubler D.J.,Vorndam A.V. et al,DEngue surveillance-United States 1986-1992, Morbidity and Mortality Weekly REport 43:7-19,1994CDC: DEngue feveramong U.S. military personnel-Haiti,September-November,1994,J.Am.Med.Assn.,273:14-15,1994Sharp T.W.,Wallace M.R.,Hayes C.G.et al,Dengue fever in US troops during Operation Restore Hope,Somalia 1992- 1993, Am.J.TRop.Med.Hyg.53:89-94,1995