Standardization of Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assays for Routine Diagnosis of Arboviral Infections

Abstract

Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) is a rapid and versatile diagnostic method that readily permits the combination of multiple assays. Test consolidation is especially important for arthropod-borne viruses (arboviruses) which belong to at least three virus families: the Togaviridae,Flaviviridae, and Bunyaviridae. Using prototype viruses from each of these families and a panel of well-characterized human sera, we have evaluated and standardized a combined MAC-ELISA capable of identifying virus infections caused by members of each virus family. Furthermore, by grouping antigens geographically and utilizing known serological cross-reactivities, we have reduced the number of antigens necessary for testing, while maintaining adequate detection sensitivity. We have determined that a 1:400 serum dilution is most appropriate for screening antiviral antibody, using a positive-to-negative ratio of ≥2.0 as a positive cutoff value. With a blind-coded human serum panel, this combined MAC-ELISA was shown to have test sensitivity and specificity that correlated well with those of other serological techniques.

Characterization and application of monoclonal antibodies specific to West Nile virus envelope protein

“…Notably, none of the four MAbs showed cross-reactivity with JEV in any of our assays. WNV and JEV are genetically closely related, and the serological cross-reactions are usually found with these viruses (Martin et al, 2000). Considering that WNV and JEV are maintained in similar transmission cycles, and have overlapping geographic distributions in Southeast Asia, MAbs which can differentiate WNV from JEV are very useful for the surveillance system in these areas.…”

“…The plaque reduction neutralization tests for typespecific diagnosis are laborious, expensive, and require live virus, which limits their application in large-scale surveillance. ELISAbased detection for IgM, IgG or IgA has been developed, and some of these assays are commercially available (Hogrefe et al, 2004;Levett et al, 2005;Martin et al, 2000;Prince and Lape-Nixon, 2005). However, the serological cross-reactions and cross-neutralizations found in the JEV serocomplex viruses limit the specificity of serological tests (Hogrefe et al, 2004;Martin et al, 2000;Niedrig et al, 2007).…”

Development and Evaluation of a SYBR Green–Based Real-Time Multiplex RT-PCR Assay for Simultaneous Detection and Serotyping of Dengue and Chikungunya Viruses

“…Enzyme-linked immunosorbent assay, especially capture of IgM, has been considered the standard for the diagnosis of arbovirus infection during the acute phase and allows diagnosis 5 to 7 days after the onset of illness but is hampered by cross-reactions between members of the Flaviviruses and Alphaviruses. 13 Shu et al 14 developed a SYBR Green Iebased real-time RT-PCR assay for DENV serotyping, but at least four tubes of reactions are needed per sample. Chien et al 15 developed a TaqMan probeebased real-time RT-PCR assay for DENV serotyping, but the method is unable to detect CHIKV simultaneously.…”

Magnetic Nanotrap Particles Preserve the Stability of Venezuelan Equine Encephalitis Virus in Blood for Laboratory Detection

“…Viral stability is an essential factor for diagnosis of VEEV to confirm the presence of virions or viral RNA in a clinical sample. The current diagnostic approaches to confirm VEEV infection in humans or horses rely on direct detection of viral nucleic acids in serum or spinal fluid samples during the acute-phase of infection using reverse-transcription PCR (RT-PCR) (11) and ELISA for VEEV-specific IgM (12). However, despite high sensitivity of the above-mentioned methods, false negative results can be obtained if samples have been collected during the initial asymptomatic phase of infection where the viral load is low (13)(14)(15).…”