HANTAVIRUS IgM ELISA
Hantavirus IgM ELISA for the qualitative determination of IgM antibodies against Hantavirus in human serum or plasma.
The qualitative immunoenzymatic determination of specific antibodies is based on the ELISA (Enzyme-linked Immunosorbent Assay) technique. Microplates are coated with specific antigens to bind corresponding antibodies of the sample. After washing the wells to remove all unbound sample material a horseradish peroxidase (HRP) labelled conjugate is added. This conjugate binds to the captured antibodies. In a second washing step unbound conjugate is removed. 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 specific antibodies in the sample. Sulphuric acid is added to stop the reaction. This produces a yellow endpoint colour. Absorbance at 450/620 nm is read using an ELISA microwell plate reader.
PRODUCT DETAILS
- Hantavirus IgM ELISA.
- High sensitivity – 96.3%.
- High specificity – 99.03%.
- Short assay time – <3 hours.
- 1 x 96 tests.
BACKGROUND
First identified in 1976 from cases of Korean hemorrhagic fever, hantaviruses are responsible for intermittent regional outbreaks of acute hemorrhagic infection. Hantaviruses are negative sense RNA viruses in the Bunyaviridae family. Humans may be infected with Hantaviruses through urine, saliva or contact with rodent waste products. Some Hantaviruses may lead to serious diseases in humans, such as hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). Human infections of Hantaviruses have almost entirely been linked to human contact with rodent excrement, but recent human-to-human transmission has been reported with the Andes virus in South America. Two clinical syndromes predominate: hemorrhagic fever renal syndrome across Europe/Asia, and hemorrhagic pulmonary syndrome in the Americas and is transmitted to humans through direct contact with rodents, excrements of rodents or aerosols. Soldiers, farmers and campers are particularly at risk. Hantavirus has an incubation time of two to four weeks in humans before symptoms of infection occur. The symptoms of HFRS can be split into five phases including Febrile phase, Hypotensive phase, Oliguric phase, Diuretic phase and finally ?a Convalescent phase when recovery occurs and symptoms begin to improve. Regions especially affected by HFRS include China, the Korean Peninsula, Russia (Hantaan, Puumala and Seoul viruses), and northern and western Europe (Puumala and Dobrava virus). A positive serological test result, evidence of viral antigen in tissue by immunohistochemistry, or the presence of amplifiable viral RNA sequences in blood or tissue, with compatible history of HPS, is considered diagnostic.
References
- Figueiredo, Luiz Tadeu Moraes; Moreli, M. L.; Borges, Alessandra Abel; Figueiredo, G. G.; Souza, R.L.M.; Aquino, V. H. (2008): Expression of a hantavirus N protein and its efficacy as antigen in immune assays. In?Braz J Med Biol Res?41 (7), pp.?596–599. DOI: 10.1590/S0100-879X2008000700008.
- Figueiredo, Luiz Tadeu Moraes; Moreli, Marcos Lazaro; Borges, Alessandra Abel; Figueiredo, Glauciane Garcia de; Badra, Soraya Jabur; Bisordi, Ivani et al. (2009): Evaluation of an enzyme-linked immunosorbent assay based on Araraquara virus recombinant nucleocapsid protein. In?The American journal of tropical medicine and hygiene?81 (2), pp.?273–276.
- Lindkvist, Marie; Naslund, Jonas; Ahlm, Clas; Bucht, Goran (2008): Cross-reactive and serospecific epitopes of nucleocapsid proteins of three hantaviruses: prospects for new diagnostic tools. In?Virus research?137 (1), pp.?97–105. DOI: 10.1016/j.virusres.2008.06.003.
- Machado, Alex M.; Machado, Aline R. S. R.; Moreli, Marcos L.; Ribeiro, Bergmann M.; Figueiredo, Luiz Tadeu Moraes; Wolff, Jose L. C. (2011): Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli. In?Virology journal?8, p.?218. DOI: 10.1186/1743-422X-8-218.
- Maes, Piet; Keyaerts, Els; Bonnet, Veronique; Clement, Jan; Avsic-Zupanc, Tatjana; Robert, Alain; van Ranst, Marc (2006): Truncated recombinant Dobrava hantavirus nucleocapsid proteins induce strong, long-lasting immune responses in mice. In?Intervirology?49 (5), pp.?253–260. DOI: 10.1159/000093454.
- Peters, C. J.; Mills, James N.; Spiropoulou, Christina; Zaki, Sherif R.; Rollin, Pierre E. (2006): Hantavirus Infections. In Richard L. Guerrant, David H. Walker, Peter F. Weller (Eds.): Tropical infectious diseases. Principles, pathogens & practice. 2nd ed. Philadelphia: Churchill Livingstone, pp.?762–780.
THIS ELISA ASSAY IS FOR RESEARCH USE ONLY. IT IS NOT FOR USE IN DIAGNOSTIC PROCEDURES.