2017 / 05

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DOI: 10.24075/brsmu.2017-05-04

METHOD

A protocol of development of a screening assay for evaluating immunological memory to vaccine-preventable infections: simultaneous detection of antibodies to measles, mumps, rubella and hepatitis B

Mazunina EP1, Kleymenov DA1, Manuilov VA1,2, Gushchin VA1,3, Tkachuk AP1
About authors

1 Laboratory of Translational Biomedicine,
N. F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Moscow, Russia

2 Institute of Molecular Medicine,
I. M. Sechenov First Moscow State Medical University, Moscow, Russia

3 Department of Virology, Faculty of Biology,
Lomonosov Moscow State University, Moscow

Correspondence should be addressed: Elena Mazunina
ul. Gamalei, d. 18, Moscow, Russia, 123098; ur.relbmar@aneleanidlan

About paper

Funding: this work was supported by the Russian Ministry of Health.

Contribution of the authors to this work: Mazunina EP, Kleymenov DA — analysis of literature, research planning, data collection, analysis, and interpretation, drafting of a manuscript, writing of a final version of a manuscript; Manuilov VA — analysis of literature, research planning, drafting of a manuscript; Gushchin VA — analysis of literature, research planning, data analysis and interpretation, drafting of a manuscript, writing of a final version of a manuscript; Tkachuk AP — analysis of literature, research planning, critical analysis of a manuscript.

Received: 2017-09-26 Accepted: 2017-10-10 Published online: 2018-01-13
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Fig. 1. Schematic representation of indirect serologic multiplex immunoassay, showing 4 bead regions, each bead coupled to a capture antigen. (A) Serum is added to the bead suspension; the first incubation is carried out. (B) Wash 1: unbound serum components are removed. (C) Phycoerythrin-conjugated anti-human detection antibodies are added to the coupled beads and the second incubation takes place. (D) Wash 2: unbound components (conjugate) are removed. (E) Beads are analyzed on the MAGPIX workstation
Fig. 2. Diagram showing steps of assay development
Fig. 3. Levels of IgG to rubella in human serum measured by xMAP and ELISA
Fig. 4. Levels of IgG to measles in human serum measured by xMAP and ELISA
Fig. 5. Levels of IgG to mumps in human serum measured by xMAP and ELISA
Fig. 6. Curves for serum pools dilutions with high antibody titers to rubella (RUB), measles (MEA), mumps (MUM) and hepatitis B (HB)
Fig. 7. Diagram illustrating the validation process of the studied assay. S1–8 — wells used to analyze calibrators and construct the calibration curve. B — wells containing a standard diluent; S+B — samples with the analyte diluted in a standard dilution buffer; S+M — samples with the analyte diluted in the matrix (rabbit serum or human serum free of antibodies)
Table 1. Antigen candidates for the assay
Table 2. Standards and serum controls used in the development of the multiplex assay
Table 3. Optimal conditions for the developed immunoassay