Development of a multiplex microsphere immunoassay for the quantitation of salivary antibody responses to selected waterborne pathogens

Griffin, Shannon M.; Chen, Ing M.; Fout, G. Shay; Wade, Timothy J.; Egorov, Andrey I.

doi:10.1016/j.jim.2010.11.005

Cited by 57 publications

(82 citation statements)

References 43 publications

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“…To measure norovirus-specific antibody responses in saliva, norovirus recombinant P particles and GST control proteins were coupled to Luminex beads (Griffin et al, 2011) or coated on MSD plates as described in Table 1. Recombinant P particles of three noroviruses, Norwalk (GI.1), VA387 (GII.4), and VA207 (GII.9), were produced using an E. coli expression system and purified as described previously (Tan and Jiang, 2005; Tan et al, 2008).…”

Section: Methodsmentioning

confidence: 99%

“…Norovirus P particles, GST, or isotype-specific anti-human capture antibodies (for total IgA and IgG tests) were covalently coupled to Luminex carboxylated microspheres as described previously (Griffin et al, 2011). Specifically, 50 mM 2-( N -morpholino) ethanesulfonic acid (MES), pH 5.0 was used as coupling buffer.…”

Section: Methodsmentioning

confidence: 99%

“…This study aimed to optimize and evaluate a previously developed approach, which employs recombinant viral proteins (P particles) for the detection of incident infections (Griffin et al, 2011). It involved further development of specifications for the Luminex-based fluorescence suspension immunoassay (Luminex Corp., Austin, TX), which was used in the previous tests, and an initial demonstration of saliva antibody tests using another common immunoassay methodology, the Meso Scale Discovery (MSD; Rockville, MD) electrochemiluminescence (ECL) platform.…”

Section: Introductionmentioning

confidence: 99%

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Application of salivary antibody immunoassays for the detection of incident infections with Norwalk virus in a group of volunteers

Griffin

Converse

León

et al. 2015

Journal of Immunological Methods

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Norovirus infection is the most common cause of acute gastroenteritis in developed countries. Developing an assay based on a non-invasive biomarker for detecting incident norovirus infections could improve disease surveillance and epidemiological investigations. This project involved analysis of IgA and IgG norovirus-specific antibody responses in saliva samples from a Norwalk virus (Genogroup I, genotype 1 norovirus) challenge study involving infected and symptomatic, and non-infected asymptomatic individuals. Saliva was collected at the challenge, and two weeks and 40 days post-challenge. Samples were analyzed using the Luminex fluorometric and Meso Scale Discovery (MSD) electrochemiluminescence immunoassays. Recombinant P domains of Norwalk virus capsid protein, as well as similar recombinant proteins of two genogroup II noroviruses (VA387 and VA207) were used as antigens. Immunoconversions were defined as >4-fold increase in antibody responses to the norovirus antigens. Various sample pre-treatment options, buffers, saliva dilution ratios, and data adjustment approaches to control for sample-to-sample variability in saliva composition were compared using the Luminex assay. The results suggest that adjusting responses to the norovirus antigens for responses to the protein purification tag, glutathione-S-transferase (GST), significantly improved the odds of producing a correct immunoconversion test result. IgG-based tests were more accurate compared to IgA-based tests. At optimal conditions, both Luminex and MSD assays for Norwalk-specific IgG antibodies correctly identified all infected and non-infected individuals. There was no evidence of cross-reactivity of anti-Norwalk virus antibodies with genogroup II noroviruses. These results suggest that salivary antibody responses can be used for the detection of incident infections with Norwalk virus in prospective surveys.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Application of salivary antibody immunoassays for the detection of incident infections with Norwalk virus in a group of volunteers

Griffin

Converse

León

et al. 2015

Journal of Immunological Methods

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“…The utility and benefits of multiplex immunoassays have been widely published. There are multiple reports of serological multiplex immunoassays for the measurement of antibodies to a wide range of pathogens, including T. gondii, Escherichia coli O157:H7, Helicobacter pylori, HIV, and noroviruses (19)(20)(21). However, multiplex immunoassays also present challenges and limitations that include cross-reactivity and assay sensitivity and specificity that must be addressed before they can be used as definitive diagnostic tools (22).…”

mentioning

confidence: 99%

Commentary: Towards Universal Screening for Toxoplasmosis: Rapid, Cost-Effective, and Simultaneous Detection of Anti-Toxoplasma IgG, IgM, and IgA Antibodies by Use of Very Small Serum Volumes

Augustine

2016

J Clin Microbiol

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“…Multiplex methods for detection of salivary antibodies have been developed and evaluated for other pathogens (16). A multiplex assay developed for measuring IgG antibodies in serum was used for quantification of anti-meningococcal serogroup C antibodies in saliva (17,18).…”

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confidence: 99%

Development and Evaluation of a Multiplex Microsphere Assay for Quantitation of IgG and IgA Antibodies against Neisseria meningitidis Serogroup A, C, W, and Y Polysaccharides

Bårnes¹,

Kristiansen²,

Caugant³

et al. 2015

Clin. Vaccine Immunol.

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We developed and evaluated a rapid and simple multiplex microsphere assay for the quantification of specific IgG and IgA antibodies against meningococcal serogroup A, C, W, and Y capsular polysaccharides in serum and saliva. Meningococcal polysaccharides were conjugated to distinct magnetic carboxylated microspheres, and the performance of the assay was assessed using the CDC1992 standard meningococcal reference serum and a panel of serum and saliva samples. The standard curve was linear over an eight 3-fold dilution range in the IgG assay and a seven 3-fold dilution range in the IgA assay. No cross-reactivity was discovered, and the assay showed high specificity with >91% homologous inhibition and <11% heterologous inhibition for all serogroups and immunoglobulin classes. Lower limits of detections were <280 pg/ml for IgG and <920 pg/ml for IgA antibodies. The assay was reproducible, with a mean coefficient of variation of <5% for intra-assay duplicates, a mean coefficient of variation of <20% for interassay repeated analysis with different conjugations of microspheres, and a mean coefficient of variation within 25.8% for interoperator variation. The assay showed good correlation to the standard meningococcal polysaccharide enzyme-linked immunosorbent assay (ELISA) for detection of serum antibodies. This multiplex assay is robust and reliable and requires less sample volume, and less time and workload are needed than for ELISA, making this method highly relevant for serological and salivary investigations on the effect of meningococcal vaccines and for immunosurveillance studies. Meningococcal disease continues to be a significant public health problem, although vaccines used in national immunization programs or mass vaccination campaigns have reduced the incidence of the disease in several countries (1). The capsular polysaccharide is an important antigen and virulence factor (2), and the most widely used meningococcal vaccines are based on these polysaccharides. Such vaccines have been shown to be effective for serogroups A, C, W, and Y, four of the five major diseasecausing meningococcal serogroups (1, 3), and have been available and widely used for nearly half a century.To evaluate the effect of meningococcal vaccines and determine protection against disease, serogroup-specific serological measures are used. Serum bactericidal activity (SBA) has become the most widely used surrogate of protection and is the basis for licensure of the recent meningococcal vaccines (4). However, this method is highly time consuming and requires specialized laboratories and highly standardized biological reagents. Quantitation of specific anti-meningococcal polysaccharide antibodies, on the other hand, is more suitable for large immunosurveillance studies and contributes to a broader understanding of the immune response. In a vaccine efficacy trial in Finland in the 1970s, a specific immunoglobulin G (IgG) concentration was shown to correlate with clinical protection against serogroup A disease (5). The most common method for ...

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Development of a multiplex microsphere immunoassay for the quantitation of salivary antibody responses to selected waterborne pathogens

Cited by 57 publications

References 43 publications

Application of salivary antibody immunoassays for the detection of incident infections with Norwalk virus in a group of volunteers

Application of salivary antibody immunoassays for the detection of incident infections with Norwalk virus in a group of volunteers

Commentary: Towards Universal Screening for Toxoplasmosis: Rapid, Cost-Effective, and Simultaneous Detection of Anti-Toxoplasma IgG, IgM, and IgA Antibodies by Use of Very Small Serum Volumes

Development and Evaluation of a Multiplex Microsphere Assay for Quantitation of IgG and IgA Antibodies against Neisseria meningitidis Serogroup A, C, W, and Y Polysaccharides

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