Development and validation of a robust multiplex serological assay to quantify antibodies specific to pertussis antigens

Rajam, Gowrisankar; Carlone, George M.; Kim, Ellie; Choi, Jin Hee; Paulos, Simon; Park, Sohee; Jeyachandran, Amilia; Gorantla, Yamini; Wong, Emily Y.; Sabnis, Amit; Browning, Peter; Desai, Rita; Quinn, Conrad P.; Schiffer, Jarad

doi:10.1016/j.biologicals.2018.11.001

Cited by 16 publications

(8 citation statements)

References 39 publications

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“…This was likely due to the unspecific binding of the protein-A label to the IgA and IgM antibodies from the samples, as a test with anti-human IgG label conjugate (Figure 3) produced no signal with the same samples from the PT test line. (16,17), (g) both PRN and FHA positive, (18,19), and (h) all negative samples (1,2,3). Please refer to Table 1 for antibody concentrations.…”

Section: Resultsmentioning

confidence: 99%

“…ELISAs are time-consuming and laborious, and pose limitations when the volume of serum samples may be limited when required to quantify the immune response to pathogens presenting multiple virulence factors. Cost-efficient and high-throughput multiplex serological assays in the pertussis field already have a broad application [16][17][18][19]. We have reported earlier on a simple, quantitative and rapid lateral flow (LF)-based platform for anti-PT IgG serological diagnostics of pertussis without the complexity of common laboratory practicalities [20].…”

Section: Introductionmentioning

confidence: 99%

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Multiplex Point-of-Care Tests for the Determination of Antibodies after Acellular Pertussis Vaccination

et al. 2020

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Most of the current serological diagnosis of pertussis is based on pertussis toxin (PT) IgG antibodies and does not differentiate between vaccination and infection-induced antibodies. PT is included in all of acellular pertussis vaccines available in the world. Multiplex testing of non-vaccine antigen-related antibodies has the potential to improve the diagnostic outcome of these assays. In this study, we developed a quantitatively spatial multiplex lateral flow immunoassay (LFIA) for the detection of IgG antibodies directed against PT, pertactin (PRN), and filamentous hemagglutinin (FHA). The assay was evaluated with serum samples with varying anti-PT, anti-PRN, and anti-FHA IgG levels and the result was compared to those obtained with standardized ELISA. The developed assay showed good specificity with PT and PRN antibodies and semiquantification throughout the antigen combinations. This exploratory study indicates that the multiplex LFIA is specific and sensitive, and a similar test platform with alternative antigens could be suitable for new type of pertussis serology.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multiplex Point-of-Care Tests for the Determination of Antibodies after Acellular Pertussis Vaccination

et al. 2020

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“…MIAs provides alternatives allowing high sensitivity, reproducibility and speci city. Several studies have reported the usefulness of multiplex platforms for immunogenicity assessment of AP-based combination vaccines 9,10,16,25,26 . A study evaluating a tetraplex microsphere assay for pertussis antigen showed high concordance with an in-house ELISA.…”

Section: Discussionmentioning

confidence: 99%

“…Multiplex immunoassays (MIA) such as Luminex x-MAP® and Meso Scale Diagnostics (MSD) offer opportunities for simultaneous quanti cation of multiple antigens in a single well. Several studies have reported the use of multiplex assays for screening antibodies against the ve vaccine antigens: tetanus toxoid (TT), diphtheria toxoid (DT), pertussis toxin (PT), lamentous hemagglutinin (FHA), and pertactin (PRN) 9,10,11 . It is important that such multiplex assays are developed and validated by reporting the results in units that are traceable to an appropriate international reference standard 12 .…”

Section: Introductionmentioning

confidence: 99%

Development and validation of bead-based assay quantifying Tetanus, Diphtheria, Pertussis Toxin, Filamentous haemagglutinin and Pertactin specific IgG in human serum

Rathod

Kadam

Gumma

et al. 2022

Preprint

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Conventional ELISA platforms have been used for vaccine immunogenicity testing. However, due to limitations in sourcing and accessibility to human serum samples, we report the development and validation of Luminex-based multiplex immunoassay (MIA), using monovalent beads, which would reduce the analysis time, cost, and sample volume while simultaneously measuring the concentration of serum immunoglobulin G (IgG) antibodies specific for tetanus (TT), diphtheria (DT), pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (PRN), using the NIBSC reference standards. Additionally, we also report the development of a multiplex reference standard (MRS) focused on the simultaneous evaluation of antibodies against T, D, PT, PRN, and FHA in healthy human sera samples. As an assay evaluation parameter, the precision, accuracy, dilutional linearity, minimum and maximum detectable limit, robustness, stability, etc were assessed. The assay exhibited a wide dynamic range for all the five antigens and could quantify the IgG concentrations down to minimum concentrations, demonstrating antigen specificity with no cross-talks among the beads. The results obtained with MIA were consistent with commercially available assays. Thus, to conclude, the study provided a pentaplex assay with increased sensitivity, reproducibility and high throughput capabilities which would allow design of large and robust clinical studies for evaluating natural and vaccine-induced immunity.

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“…Defining the Ab function/s and thresholds that can accurately predict vaccine performance could accelerate the evaluation of improved candidates and guide implementation. New assays have been applied for high throughout measurements of B. pertussis-specific Abs (34). However, most studies continue to rely primarily on Ab binding, providing limited new knowledge on their actual in vivo effects on various aspects of infection.…”

Section: Adjuvants To Elicit Persistent Immunological Memorymentioning

confidence: 99%

Overcoming Waning Immunity in Pertussis Vaccines: Workshop of the National Institute of Allergy and Infectious Diseases

Damron

Barbier

Dubey

et al. 2020

The Journal of Immunology

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Despite high vaccine coverage in many parts of the world, pertussis is resurging in a number of areas in which acellular vaccines are the primary vaccine administered to infants and young children. This is attributed in part to the suboptimal and short-lived immunity elicited by acellular pertussis vaccines and to their inability to prevent nasal colonization and transmission of the etiologic agent Bordetella pertussis. In response to this escalating public health concern, the National Institute of Allergy and Infectious Diseases held the workshop “Overcoming Waning Immunity in Pertussis Vaccines” in September 2019 to identify issues and possible solutions for the defects in immunity stimulated by acellular pertussis vaccines. Discussions covered aspects of the current problem, gaps in knowledge and possible paths forward. This review summarizes presentations and discussions of some of the key points that were raised by the workshop.

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Development and validation of a robust multiplex serological assay to quantify antibodies specific to pertussis antigens

Cited by 16 publications

References 39 publications

Multiplex Point-of-Care Tests for the Determination of Antibodies after Acellular Pertussis Vaccination

Multiplex Point-of-Care Tests for the Determination of Antibodies after Acellular Pertussis Vaccination

Development and validation of bead-based assay quantifying Tetanus, Diphtheria, Pertussis Toxin, Filamentous haemagglutinin and Pertactin specific IgG in human serum

Overcoming Waning Immunity in Pertussis Vaccines: Workshop of the National Institute of Allergy and Infectious Diseases

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