HANTAVIRUS IGG ELISA

Hantavirus IgG ELISA for the detection and qualitative determination of 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 IGG ELISA

• Hantavirus IgG ELISA.
• High sensitivity – 99.16%.
• High specificity – 96.59%.
• Short assay time – <3 hours.
• 1 x 96 tests.

BACKGROUND

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. 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:

1. Febrile phase: Symptoms include fever, chills, sweaty palms, diarrhea, malaise, headaches, nausea, abdominal and back pain, respiratory problems such as the ones common in influenza virus infection, as well as gastro-intestinal problems. These symptoms normally occur for three to seven days and arise about two to three weeks after exposure.
2. Hypotensive phase: This occurs when the blood platelet levels drop and symptoms can lead to tachycardia and hypoxemia. This phase can last for 2 days.
3. Oliguric phase: This phase lasts for three to seven days and is characterized by the onset of renal failure and proteinuria occurs.
4. Diuretic phase: This is characterized by diuresis of three to six liters per day, which can last for a couple of days up to weeks.
5. Convalescent phase: This is normally 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).

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.