Validation of IgG-sandwich and IgM-capture ELISA for the detection of antibody to Rift Valley fever virus in humans

Rift Valley fever (RVF) virus is an important zoonotic and a potential biothreat agent. This paper describes validation of sandwich and capture enzyme-linked immunoassays (ELISA) based on gamma-irradiated antigens for the detection of RVFV-specific IgG and IgM antibody in humans. Validation data set...

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Bibliographic Details
Published in:Journal of virological methods Vol. 124; no. 1; pp. 173 - 181
Main Authors: Paweska, Janusz T., Burt, Felicity J., Swanepoel, Robert
Format: Journal Article
Language:English
Published: London Elsevier B.V 01.03.2005
Amsterdam Elsevier
New York, NY
Subjects:
Antibodies, Viral - blood
Biological and medical sciences
ELISA
Enzyme-Linked Immunosorbent Assay
Fundamental and applied biological sciences. Psychology
Humans
IgG antibody
IgM antibody
Immunoglobulin G - blood
Immunoglobulin M - blood
Microbiology
Rift Valley fever virus
Rift Valley fever virus - immunology
Techniques used in virology
Virology
Rift Valley fever virus
IgG antibody
IgM antibody
Humans
ELISA
Human
Microbiology
Antibody
Sandfly fever group virus
IgG
Method
IgM
Rift valley fever virus
Virology
Virus
Bunyaviridae
Detection
ELISA assay
Phlebovirus
ISSN:0166-0934, 1879-0984
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Summary:Rift Valley fever (RVF) virus is an important zoonotic and a potential biothreat agent. This paper describes validation of sandwich and capture enzyme-linked immunoassays (ELISA) based on gamma-irradiated antigens for the detection of RVFV-specific IgG and IgM antibody in humans. Validation data sets derived from testing field-collected sera from Africa ( n = 2400) were dichotomised according to the results of a virus neutralisation test. In addition, sera from laboratory workers immunized with inactivated RVF vaccine ( n = 93) and serial sera ( n = 3) from a single RVF case were used. ELISA data were expressed as percentage of high-positive control serum (PP). Cut-off values at 95% accuracy level were optimised using the misclassification cost term option of the two-graph receiver operating characteristics analysis. During the routine use of assays there was no evidence for excessive intra- and inter-plate variations within and between runs of assays. At a cut-off of 13.2 PP the sensitivity of the IgG-sandwich ELISA was 100% and specificity 99.95%, while for the IgM-capture ELISA the values were 96.47 and 99.44%, respectively, at a cut-off of 7.1 PP. Compared to the virus neutralisation test, the IgG-sandwich ELISA was more sensitive in detection of immunological responses in vaccines. Following natural infection class-specific antibodies were detected in serum taken 6 days after onset of symptoms. The results demonstrate that both assays will be useful for early diagnosis of infection, epidemiological surveillance and for monitoring of immune response after vaccination. As highly accurate, robust and safe tests, they have the potential to replace traditional diagnostic methods which are unable to distinguish between different classes of immunoglobulins, and pose health risks necessitating their use being restricted to high containment facilities outside RVF endemic areas.
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ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2004.11.020