Multiplexed immunoassay to assess Shigella-specific antibody responses

Kaminski, Robert W.; Clarkson, Kristen A.; Kordis, Alexis A.; Oaks, Edwin V.

doi:10.1016/j.jim.2013.04.002

Cited by 12 publications

(11 citation statements)

References 43 publications

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“…The similarity between the multiplex and singleplex assays also reinforces the good pattern of specificity demonstrated in the inhibition of binding studies, as the few cases of crossreactivity did not persist when the multiplex assay was performed using the standard protocol. Finally, taking into consideration that previous studies considered an unspecific background as the one with MFI values N 250 (Kaminski et al, 2013;Verkaik et al, 2008;Waterboer et al, 2006), an importantly low unspecific binding (MFI b20) was found with the uncoupled beads in this assay. Therefore, we conclude that, overall, the developed assay demonstrated good specificity.…”

Section: Discussionmentioning

confidence: 95%

“…Comparison with ELISA has been the routine procedure in the validation of FMIA protocols (Kaminski et al, 2013;Lal et al, 2005;Pickering et al, 2002a;Pickering et al, 2002b;van Gageldonk et al, 2008) and the absence of a comparison between the FMIA and ELISA for the antigens from H. influenzae and M. catarrhalis represents another limitation of our study.…”

Section: Plymentioning

confidence: 99%

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A fluorescent multiplexed bead-based immunoassay (FMIA) for quantitation of IgG against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis protein antigens

Andrade

Borges

Laitinen

et al. 2014

Journal of Immunological Methods

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Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are pathogens commonly associated with infectious diseases in childhood. This study aimed to develop a fluorescent multiplexed bead-based immunoassay (FMIA) using recombinant proteins for the quantitation of serum IgG antibodies against these bacteria. Eight pneumococcal proteins (Ply, CbpA, PspA1, PspA2, PcpA, PhtD, SP1732-3 and SP2216-1), 3 proteins of H. influenzae (NTHi Protein D, NTHi0371-1, NTHi0830), and 5 proteins of M. catarrhalis (MC Omp CD, MC_RH4_2506, MC_RH4_1701, MC_RH4_3729-1, MC_RH4_4730) were used to develop the FMIA. Optimal coupling concentrations for each protein, comparison of singleplex and multiplex assays, specificity, reproducibility, and correlation to ELISA for six pneumococcal antigens were determined for validation. FMIA was then used to analyze acute and convalescent paired serum samples of 50 children with non-severe pneumonia. The coupling concentrations varied for different antigens, ranging from 1.6 to 32μg of protein/million beads. Correlation between singleplexed and multiplexed assays was excellent, with R≥0.987. The FMIA was specific, reaching >92% homologous inhibition for all specificities; heterologous inhibition ≥20% was found only in six cases. The assay was repeatable, with averages of intra-assay variation ≤10.5%, day-to-day variation ≤9.7% and variation between technicians ≤9.1%. Comparison with ELISA for pneumococcal antigens demonstrated good correlation with R ranging from 0.854 (PspA2) to 0.976 (PcpA). The samples from children showed a wide range of antibody concentrations and increases in convalescent samples. In conclusion, the FMIA was sensitive, specific, and repeatable, using small amounts of recombinant proteins and sera to detect antibodies against S. pneumoniae, H. influenzae and M. catarrhalis. The methodology would be suitable for studies investigating etiological diagnosis and in experimental vaccine studies.

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Section: Discussionmentioning

confidence: 95%

Section: Plymentioning

confidence: 99%

A fluorescent multiplexed bead-based immunoassay (FMIA) for quantitation of IgG against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis protein antigens

Andrade

Borges

Laitinen

et al. 2014

Journal of Immunological Methods

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“…Levels of IL-6 (Mouse IL-6 Quantikine #LXSAMSM, R&D systems, M.N., U.S.) and TNF-alpha (Mouse TNF-alpha Quantikine #LXSAMSM, R&D systems, M.N., U.S.) in plasma were measured by ELISA. Blood samples were collected from mice via the cardiac ventricle and heparin serum plasma was stored at -80 °C until analysis 14-16.…”

Section: Methodsmentioning

confidence: 99%

Positive Prehabilitative Effect of Intense Treadmill Exercise for Ameliorating Cancer Cachexia Symptoms in a Mouse Model

Jee¹,

Chang

Yang³

2016

J. Cancer

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Due to the importance of exercise in prehabilitation, we conducted this study to understand the effects of different exercise intensities on cancer-related cachexia.Forty adult male CDF1 mice were randomly divided into a non-cancer control group (N=10, NC), cancer control group (N=10, CC), cancer with moderate exercise group (N=10, ME, 70% maxHR), and cancer with intense exercise group (N=10, SE, 90% maxHR) for obtaining data such as tissue weight and body weight changes, quality of life (QoL) indicators, and levels of cytokines and a muscle homeostasis regulatory protein.We verified that mouse colonic carcinoma cancer cells metastasized based on our observation that the weight of CC group lungs was almost 87% greater than NC group lungs. Survival rates of SE, NC, ME, and CC groups were 100%, 100%, 80%, and 50%, respectively (p<0.01). Other results such as tissue and body weight changes, QoL indicators, and protein analyses also supported our hypothesis that the SE group had improved survival compared to CC and ME groups (p<0.05 and p<0.01, respectively).Our results suggest that exercise, especially intense exercise, improves QoL and survival rate and prevents muscle atrophy. These data suggest that exercise is an optimal prehabilitation choice to alleviate the negative impacts of cancer cachexia.

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“…Interestingly, arthritogenicity of Shigella flexneri 2a was not associated with the presence or absence of antibodies toward epitopes of the cell wall (49). Advanced Lumixex™ technology allowed simultaneous detection of specific antibodies to recombinant invasin plasmid antigens Ipa B, C, and D as wells as to LPS from Shigella sonnei, flexneri 2a , and dysenteriae (50). Although promising, this technology has not yet been widely evaluated to support ReA diagnosis.…”

Section: Serology For Shigella Infectionsmentioning

confidence: 99%

A Review of Serological Tests to Assist Diagnosis of Reactive Arthritis: Critical Appraisal on Methodologies

Tuuminen

Lounamo²,

Leirisalo‐Repo

2013

Front. Immunol.

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On a population-based level, the incidence of reactive arthritis (ReA) is 0.6–27/100,000. The definition of ReA varies and its pathogenesis is not yet clear. Attempts in basic immunology to suggest hypotheses for proliferation of forbidden B cell clones, molecular mimicry, and involvement of cross-reactive antibodies are helpful but not sufficient. Importantly, for the clinical diagnosis of the preceding infection, serology is widely used. Unfortunately, the accuracy of associations between serologic findings and clinical conclusions is plagued by poor standardization of methods. So far, few attempts have been done to examine the pitfalls of different approaches. Here, we review several serologic techniques, their performance and limitations. We will focus on serology for Yersinia, Campylobacter, Salmonella, Shigella, and Chlamydia trachomatis because these bacteria have a longer history of being associated with ReA. We also address controversies regarding the role of serology for some other bacteria linked to autoimmune disorders.

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Multiplexed immunoassay to assess Shigella-specific antibody responses

Cited by 12 publications

References 43 publications

A fluorescent multiplexed bead-based immunoassay (FMIA) for quantitation of IgG against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis protein antigens

A fluorescent multiplexed bead-based immunoassay (FMIA) for quantitation of IgG against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis protein antigens

Positive Prehabilitative Effect of Intense Treadmill Exercise for Ameliorating Cancer Cachexia Symptoms in a Mouse Model

A Review of Serological Tests to Assist Diagnosis of Reactive Arthritis: Critical Appraisal on Methodologies

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