Preliminary Communication

Validation and comprehensive analysis of Streptococcus pneumoniae IgG WHO enzyme-linked immunosorbent assay in an Indian reference laboratory

https://orcid.org/0000-0001-6473-1724

Central Research Laboratory, Kempegowda Institute of Medical Sciences, Bangalore, India

Department of Biotechnology and Genetics, School of Sciences, JAIN (Deemed-to-be University), Bangalore, India

Search for more papers by this author

Govindan Vandana

Central Research Laboratory, Kempegowda Institute of Medical Sciences, Bangalore, India

Search for more papers by this author

Manheri Mavupadi Akhila

Central Research Laboratory, Kempegowda Institute of Medical Sciences, Bangalore, India

Search for more papers by this author

Ravindran Shilpa

Central Research Laboratory, Kempegowda Institute of Medical Sciences, Bangalore, India

Search for more papers by this author

Kadahalli Lingegowda Ravikumar

*Author for correspondence:

E-mail Address: klravikumar@gmail.com

https://orcid.org/0000-0001-8629-1459

Central Research Laboratory, Kempegowda Institute of Medical Sciences, Bangalore, India

Search for more papers by this author

Published Online:5 Feb 2024https://doi.org/10.4155/bio-2023-0111

View article

Abstract

Monitoring serotype-specific IgG levels against pneumococci is crucial for assessing immunity, vaccine efficacy, and evaluating vaccination programs. The WHO ELISA for pneumococci is a standardized assay ensuring consistency in testing and comparability of results across laboratories. It involves a rigorous testing process to confirm accurate, precise and reliable detection of antibodies. We validated the protocol for 13 pneumococcal serotypes by assessing its specificity, reproducibility (coefficient of variation ≤15%), repeatability (coefficient of variation ≤20%), accuracy, lower limit of quantification, stability, and robustness. We found these parameters were within acceptable ranges and showed excellent performance. Our findings imply that the method employed is appropriate for evaluating 13 valent pneumococcal conjugate vaccine which is introduced in the national immunization program by comparing pre-and post-vaccination IgG response.

Keywords:

Papers of special note have been highlighted as: • of interest; •• of considerable interest

References

1. Weiser JN, Ferreira DM, Paton JC. Streptococcus pneumoniae: transmission, colonization and invasion. Nat. Rev. Microbiol. 16(6), 355–367 (2018). • This comprehensive review provides insights into the transmission and pathogenesis of Streptococcus pneumoniae that are crucial for understanding the disease.
Medline CASGoogle Scholar
2. Henrichsen J. Six newly recognized types of Streptococcus pneumoniae. J. Clin. Microbiol. 33(10), 2759–2762 (1995). • This seminal work identifies and characterizes new S. pneumoniae types, contributing to the understanding of serotype diversity.
Medline CASGoogle Scholar
3. Ganaie F, Saad JS, McGee L et al. A new pneumococcal capsule type, 10D, is the 100th serotype and has a large cps fragment from an oral Streptococcus. mBio 11(3), e00937–20 (2020). • Presents the discovery of a new pneumococcal capsule type, highlighting the ongoing evolution of the pathogen.
MedlineGoogle Scholar
4. Brooks LRK, Mias GI. Streptococcus pneumoniae‘s virulence and host immunity: aging, diagnostics, and prevention. Front. Immunol. 9, 1366 (2018).
MedlineGoogle Scholar
5. Wright PF, Nilsson E, Van Rooij EM, Lelenta M, Jeggo MH. Standardisation and validation of enzyme-linked immunosorbent assay techniques for the detection of antibody in infectious disease diagnosis. Rev. Sci. Tech. 12(2), 435–450 (1993). • Provides valuable information on the standardization and validation of ELISA techniques for infectious disease diagnosis.
Medline CASGoogle Scholar
6. Sorensen RU, Edgar JDM. Overview of antibody-mediated immunity to S. pneumoniae: pneumococcal infections, pneumococcal immunity assessment, and recommendations for IG product evaluation. Transfusion 58(Suppl. 3), 3106–3113 (2018). • This overview addresses antibody-mediated immunity to Streptococcus pneumoniae, offering insights for product evaluation.
Medline CASGoogle Scholar
7. Wernette CM, Frasch CE, Madore D et al. Enzyme-linked immunosorbent assay for quantitation of human antibodies to pneumococcal polysaccharides. Clin. Diagn. Lab. Immunol. 10(4), 514–519 (2003). • Outlines an ELISA for quantitating human antibodies to pneumococcal polysaccharides.
Medline CASGoogle Scholar
8. Marchese RD, Jain NT, Antonello J et al. Enzyme-linked immunosorbent assay for measuring antibodies to pneumococcal polysaccharides for the PNEUMOVAX 23 vaccine: assay operating characteristics and correlation to the WHO international. Clin. Vaccine Immunol. 13(8), 905–912 (2006).
Medline CASGoogle Scholar
9. WHO Pneumococcal Serology Reference Laboratories. Training manual for enzyme linked immunosorbent assay for the quantitation of Streptococcus pneumoniae serotype specific IgG (Pn PS ELISA) (007sp version). WHO, Geneva, Switzerland. ••References 9 and 10 provide essential guidelines and methods for the quantitation of S. pneumoniae serotype-specific IgG, serving as valuable resources for standardized serological testing in the context of pneumococcal research and vaccine assessment.
Google Scholar
10. Burd EM. Validation of laboratory-developed molecular assays for infectious diseases. Clin. Microbiol. Rev. 23(3), 550–576 (2010).
Medline CASGoogle Scholar
11. Goldblatt D, Plikaytis BD, Akkoyunlu M et al. Establishment of a new human pneumococcal standard reference serum, 007sp. Clin. Vaccine Immunol. 18(10), 1728–1736 (2011).
Medline CASGoogle Scholar
12. Medicines & Healthcare products Regulatory Agency (2017). www.nibsc.org/documents/ifu/12-278.pdf
Google Scholar
13. Lee H, Lim SY, Kim KH. Validation of the World Health Organization enzyme-linked immunosorbent assay for the quantitation of immunoglobulin G serotype-specific anti-pneumococcal antibodies in human serum. J. Korean Med. Sci. 32(10), 1581–1587 (2017). •• References 13 and 14 validated the WHO's ELISA method for quantifying serotype-specific antipneumococcal antibodies against seven serotypes in human serum, offering important insights into the assay's performance and suitability for serological testing in the context of pneumococcal research.
Medline CASGoogle Scholar
14. Minic R, Zivkovic I. Chapter optimization, validation and standardization of ELISA. Norovirus (peer-reviewed chapter).
Google Scholar
15. Guidance for Industry: Bioanalytical Method Validation. US FDACenter for Drug Evaluation and Research,Center for Veterinary Medicine. Silver Spring, MD, USA (2001).
Google Scholar
16. Wright PF, Nilsson E, Van Rooij EM, Lelenta M, Jeggo MH. Standardisation and validation of enzyme-linked immunosorbent assay techniques for the detection of antibody in infectious disease diagnosis. Rev. Sci. Tech. 12(2), 435–450 (1993). •• References 16 and 17 validated the WHO's ELISA method for quantifying serotype-specific antipneumococcal antibodies against seven serotypes in human serum, offering important insights into the assay's performance and suitability for serological testing in the context of pneumococcal research.
Medline CASGoogle Scholar
17. Belmonti S, Lombardi F, Morandi M et al. Evaluation and optimization of an ELISA procedure to quantify antibodies against pneumococcal polysaccharides included in the 13-valent conjugate vaccine. J. Immunoassay Immunochem. 37(2), 189–200 (2016).
Medline CASGoogle Scholar
18. Validation of Analytical Procedures: Text and Methodology Q2(R1). International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf
Google Scholar
19. Goldblatt D, Levinsky RJ, Turner MW. Role of cell wall polysaccharide in the assessment of IgG antibodies to the capsular polysaccharides of Streptococcus pneumoniae in childhood. J. Infect. Dis. 166(3), 632–634 (1992).
Medline CASGoogle Scholar
20. Yu X, Sun Y, Frasch C, Concepcion N, Nahm MH. Pneumococcal capsular polysaccharide preparations may contain non-C-polysaccharide contaminants that are immunogenic. Clin. Diagn. Lab. Immunol. 6(4), 519–524 (1999).
Medline CASGoogle Scholar
21. Balmer P, North J, Baxter D et al. Measurement and interpretation of pneumococcal IgG levels for clinical management. Clin. Exp. Immunol. 133(3), 364–369 (2003).
Medline CASGoogle Scholar
22. Concepcion NF, Frasch CE. Pneumococcal type 22f polysaccharide absorption improves the specificity of a pneumococcal-polysaccharide enzyme-linked immunosorbent assay. Clin. Diagn. Lab. Immunol. 8(2), 266–272 (2001).
Medline CASGoogle Scholar
23. WHO Expert Committee on Biological Standardization. Recommendations to assure the quality, safety and efficacy of pneumococcal conjugate vaccines. Geneva, Switzerland, Annex 3 Recommendations to assure the quality, safety and efficacy of pneumococcal conjugate vaccines Replacement of WHO Technical Report Series, No. 927, Annex 2. 19–23 October 2009.
Google Scholar
24. Schuerman L, Wysocki J, Tejedor JC, Knuf M, Kim KH, Poolman J. Prediction of pneumococcal conjugate vaccine effectiveness against invasive pneumococcal disease using opsonophagocytic activity and antibody concentrations determined by enzyme-linked immunosorbent assay with 22F adsorption. Clin. Vaccine Immunol. 18(12), 2161–2167 (2011).
Medline CASGoogle Scholar
25. Park DE, Johnson TS, Nonyane BA et al. The differential impact of coadministered vaccines, geographic region, vaccine product and other covariates on pneumococcal conjugate vaccine immunogenicity. Pediatr. Infect. Dis. J. 33(Suppl. 2), S130–S139 (2014).
MedlineGoogle Scholar

Vol. 16, No. 4

Supplemental Materials

Metrics

Downloaded 79 times

History

Received 26 May 2023

Accepted 19 January 2024

Published online 5 February 2024

Published in print February 2024

Information

Keywords

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.future-science.com/doi/suppl/10.4155/bio-2023-0111

Author contributions

MR Shincy conducted the study and analyzed and interpreted the data. V Govindan designed the study and contributed to manuscript writing. MM Akhila and S Ravindran helped run the assays. KL Ravikumar revised the manuscript. All authors read and approved the final manuscript.

Acknowledgments

Rijo Joseph John (Instruments Manager), G K Netravathy (Accounts Manager), and K N Ravishankar (Accounts Manager) of the Central Research Laboratory, Kempe Gowda Institute of Medical Sciences, Bengaluru, India.

Financial disclosure

This work was supported by funding from the Biotechnology Industry Research Assistance Council (BIRAC), Government of India under award no. BT/NBM/0097/02/18 with a project entitled “Establishment of Pneumococcal Vaccine Immunogenicity Evaluation Center at CRL-Kempegowda Institute of Medical Sciences, Bangalore” under the Industry-Academia Collaborative Mission for Accelerating Discovery Research to Early Development for Biopharmaceuticals. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

PDF download