Improved specificity and reduced subtype cross-reactivity for antibody detection by ELISA using globular head domain recombinant hemagglutinin

Li, Zhu-Nan; Carney, P.J.; Lin, Seh-Ching; Li, Ji; Chang, Jae C.; Veguilla, Vic; Stevens, James; Miller, Joseph D.; Levine, Min Z.; Katz, Jacqueline M.; Hancock, Kathy

doi:10.1016/j.jviromet.2014.09.006

Cited by 7 publications

(13 citation statements)

References 27 publications

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“…16,17 The use of GH HA1 rHAs that lack the antigenically conserved HA stalk can reduce such cross-reactivity. 18 In this study, we observed high MFIs against We have previously demonstrated that cross-reactivity could be reduced by incorporating a serum adsorption step prior to the MAGPIX or MN assays, and the remaining signals from post-adsorption samples represent HA subtype-specific responses. 4,7 Thus, serum adsorptions were performed in the current study to remove cross-reactive antibodies to improve sensitivity and specificity.…”

Section: Serum Adsorption By Using Either 2 or 8 Rhas From A(h1n1) supporting

confidence: 50%

“…On the other hand, in the S2 sera collected from poultry workers exposed to A(H5N1), MFIs against seasonal H1 and H3 were reduced for both the MN ≥40 and MN <40 groups after 2‐Ads (Figure F,G), but only MN <40 group reached statistically significant difference (Figure G). MFIs against H5.Ind.05 Ecto and H7.NY.16 Ecto domain rHAs in the MN <40 group were significantly reduced after 2‐Ads, but not for antibodies against H5.Ind.05 GH (Figure G), suggesting antibodies against GH HA1 are more strain‐specific, while antibodies against the whole ectodomain containing the HA stalk region are more cross‐reactive …”

Section: Resultsmentioning

confidence: 97%

“…In general, antibodies to novel virus HA in unexposed populations increase with age and vary by geographical location due to cross‐reactivity between the novel virus and seasonal influenza viruses through past exposure . When A(H1N1)pdm09 virus was first introduced to the human population in 2009, higher MN titers were observed in older age groups .…”

Section: Resultsmentioning

confidence: 99%

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Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

Cheng

Poirot

et al. 2019

Influenza Resp Viruses

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Background The development of serologic assays that can rapidly assess human exposure to novel influenza viruses remains a public health need. Previously, we developed an 11‐plex magnetic fluorescence microsphere immunoassay (MAGPIX) by using globular head domain recombinant hemagglutinins (rHAs) with serum adsorption using two ectodomain rHAs. Methods We compared sera collected from two cohorts with novel influenza exposures: animal shelter staff during an A(H7N2) outbreak in New York City in 2016‐2017 (n = 119 single sera) and poultry workers from a live bird market in Bangladesh in 2012‐2014 (n = 29 pairs). Sera were analyzed by microneutralization (MN) assay and a 20‐plex MAGPIX assay with rHAs from 19 influenza strains (11 subtypes) combined with serum adsorption using 8 rHAs from A(H1N1) and A(H3N2) viruses. Antibody responses were analyzed to determine the novel influenza virus exposure. Results Among persons with novel influenza virus exposures, the median fluorescence intensity (MFI) against the novel rHA from exposed influenza virus had the highest correlation with MN titers to the same viruses and could be confirmed by removal of cross‐reactivity from seasonal H1/H3 rHAs following serum adsorption. Interestingly, in persons with exposures to novel influenza viruses, age and MFIs against exposed novel HA were negatively correlated, whereas in persons without exposure to novel influenza viruses, age and MFI against novel HAs were positively correlated. Conclusions This 20‐plex high‐throughput assay with serum adsorption will be a useful tool to detect novel influenza virus infections during influenza outbreak investigations and surveillance, especially when well‐paired serum samples are not available.

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Section: Serum Adsorption By Using Either 2 or 8 Rhas From A(h1n1) supporting

confidence: 50%

Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

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Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

Cheng

Poirot

et al. 2019

Influenza Resp Viruses

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Section: Discussionmentioning

confidence: 99%

“…As a result of exposure(s) to seasonal influenza viruses and/or vaccine(s), antibodies to novel HA in unexposed populations increases with age and may vary by geographic location. 3,[18][19][20][21] People who were born before 1967 showed significantly higher reactivity against H2 GH HA1 than those born after 1970 (P<.05) and could not be removed by serum adsorption. They also showed higher antibody baselines (S1) against other group 1 HAs, such as, pH1, H5 VN, and H9 due to potential exposure to H2N2 virus and more seasonal H1N1 viruses.…”

Section: Discussionmentioning

confidence: 99%

Novel multiplex assay platforms to detect influenza A hemagglutinin subtype‐specific antibody responses for high‐throughput and in‐field applications

Trost

Weber

et al. 2017

Influenza Resp Viruses

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BackgroundDetections of influenza A subtype‐specific antibody responses are often complicated by the presence of cross‐reactive antibodies. We developed two novel multiplex platforms for antibody detection. The multiplexed magnetic fluorescence microsphere immunoassay (MAGPIX) is a high‐throughput laboratory‐based assay. Chembio Dual Path Platform (DPP) is a portable and rapid test that could be used in the field.MethodsTwelve recombinant globular head domain hemagglutinin (GH HA1) antigens from A(H1N1)pdm09 (pH1N1), A(H2N2), A(H3N2), A(H5N1), A(H7N9), A(H9N2), A(H13N9), B/Victoria lineage, B/Yamagata lineage viruses, and protein A control were used. Human sera from U.S. residents either vaccinated (with H5N1 or pH1N1) or infected with pH1N1 influenza viruses and sera from live bird market workers in Bangladesh (BDPW) were evaluated. GH HA1 antigens and serum adsorption using full ectodomain recombinant hemagglutinins from A(pH1N1) and A(H3N2) were introduced into the platforms to reduce cross‐reactivity.ResultsSerum adsorption reduced cross‐reactivity to novel subtype HAs. Compared to traditional hemagglutination inhibition or microneutralization assays, when serum adsorption and the highest fold rise in signals were used to determine positivity, the correct subtype‐specific responses were identified in 86%‐100% of U.S. residents exposed to influenza antigens through vaccination or infection (N=49). For detection of H5N1‐specific antibodies in sera collected from BDPW, H5 sensitivity was 100% (six of six) for MAGPIX, 83% (five of six) for DPP, H5 specificity was 100% (15/15), and cross‐reactivity against other subtype was 0% (zero of six) for both platforms.ConclusionMAGPIX and DPP platforms can be utilized for high‐throughput and in‐field detection of novel influenza virus infections.

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Influenza A H1N1 Virus 2009 Synthetic Hemagglutinin and Neuraminidase Peptides for Antibody Detection

Ávila

Cruz-Licea

Rojas-Espinosa

et al. 2020

Archives of Medical Research

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Improved specificity and reduced subtype cross-reactivity for antibody detection by ELISA using globular head domain recombinant hemagglutinin

Cited by 7 publications

References 27 publications

Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

Novel multiplex assay platforms to detect influenza A hemagglutinin subtype‐specific antibody responses for high‐throughput and in‐field applications

Influenza A H1N1 Virus 2009 Synthetic Hemagglutinin and Neuraminidase Peptides for Antibody Detection

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