Determination of avidity of IgG against protein antigens from Streptococcus pneumoniae: assay development and preliminary application in clinical settings

  • D. C. Andrade
  • I. C. Borges
  • N. Ekström
  • T. Jartti
  • T. Puhakka
  • A. Barral
  • H. Kayhty
  • O. Ruuskanen
  • C. M. Nascimento-Carvalho
Original Article


The measurement of antibody levels is a common test for the diagnosis of Streptococcus pneumoniae infection in research. However, the quality of antibody response, reflected by avidity, has not been adequately evaluated. We aimed to evaluate the role of avidity of IgG against eight pneumococcal proteins in etiologic diagnosis. Eight pneumococcal proteins (Ply, CbpA, PspA1 and 2, PcpA, PhtD, StkP-C, and PcsB-N) were used to develop a multiplex bead-based avidity immunoassay. The assay was tested for effects of the chaotropic agent, multiplexing, and repeatability. The developed assay was applied to paired samples from children with or without pneumococcal disease (n = 38 for each group), determined by either serology, polymerase chain reaction (PCR), or blood culture. We found a good correlation between singleplex and multiplex assays, with r ≥ 0.94.The assay was reproducible, with mean inter-assay variation ≤ 9% and intra-assay variation < 6%. Children with pneumococcal disease had lower median avidity indexes in the acute phase of disease for PspA1 and 2 (p = 0.042), PcpA (p = 0.002), PhtD (p = 0.014), and StkP-C (p < 0.001). When the use of IgG avidity as a diagnostic tool for pneumococcal infection was evaluated, the highest discriminative power was found for StkP-C, followed by PcpA (area under the curve [95% confidence interval, CI]: 0.868 [0.759–0.977] and 0.743 [0.607–879], respectively). The developed assay was robust and had no deleterious influence from multiplexing. Children with pneumococcal disease had lower median avidity against five pneumococcal proteins in the acute phase of disease compared to children without disease.



We thank Sanofi Pasteur (Lyon, France) for supplying PcpA and PhtD; Prof. Elaine Tuomanen at St. Jude Children’s Research Hospital (Memphis, TN, USA) for supplying Ply, CbpA, and PspA1; Profs. Susan Hollingshead, David Briles, and Pat Coan at University of Alabama (Birmingham, AL, USA) for supplying PspA2; and Valneva Austria GmbH (Vienna, Austria) for supplying StkP-C and PcsB-N.

Data availability statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.


This work was supported by: Bahia State Agency for Research Funding (FAPESB), Brazil; Brazilian Council for Scientific and Technological Development (CNPq), Brazil; Turku University Hospital Research Foundation, Finland; Rauno and Anne Puolimatka Foundation, Finland; Sohlberg Foundation, Finland.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The use of the samples was approved by the Ethics Committee of the Federal University of Bahia in Brazil, the Ethics Committee of the National Institute for Health and Welfare in Finland (formerly National Public Health Institute), and the Ethics Committee of Satakunta Central Hospital, Pori, Finland.

Informed consent

Written informed consent was obtained from legal guardians before recruitment.


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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • D. C. Andrade
    • 1
  • I. C. Borges
    • 1
  • N. Ekström
    • 2
  • T. Jartti
    • 3
  • T. Puhakka
    • 4
    • 5
  • A. Barral
    • 6
  • H. Kayhty
    • 2
  • O. Ruuskanen
    • 3
  • C. M. Nascimento-Carvalho
    • 7
  1. 1.Postgraduate Programme in Health SciencesFederal University of Bahia School of MedicineSalvadorBrazil
  2. 2.National Institute for Health and WelfareHelsinkiFinland
  3. 3.Department of PaediatricsUniversity of Turku and Turku University HospitalTurkuFinland
  4. 4.Department of OtorhinolaryngologyUniversity of Turku and Turku University HospitalTurkuFinland
  5. 5.Department of OtorhinolaryngologySatakunta Central HospitalPoriFinland
  6. 6.Pathology Department and Postgraduate Programme in Health SciencesFederal University of Bahia School of Medicine and Centro de Pesquisa Gonçalo Muniz, Fundação Oswaldo CruzSalvadorBrazil
  7. 7.Department of Pediatrics and Postgraduate Programme in Health SciencesFederal University of Bahia School of MedicineSalvadorBrazil

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