J Korean Med Sci.  2017 Oct;32(10):1581-1587. 10.3346/jkms.2017.32.10.1581.

Validation of the World Health Organization Enzyme-Linked Immunosorbent Assay for the Quantitation of Immunoglobulin G Serotype-Specific Anti-Pneumococcal Antibodies in Human Serum

• Lee H
• Lim SY
• Kim KH

• KMID: 2400428
• DOI: http://doi.org/10.3346/jkms.2017.32.10.1581

Abstract

The World Health Organization (WHO) enzyme-linked immunosorbent assay (ELISA) guideline is currently accepted as the gold standard for the evaluation of immunoglobulin G (IgG) antibodies specific to pneumococcal capsular polysaccharide. We conducted validation of the WHO ELISA for 7 pneumococcal serotypes (4, 6B, 9V, 14, 18C, 19F, and 23F) by evaluating its specificity, precision (reproducibility and intermediate precision), accuracy, spiking recovery test, lower limit of quantification (LLOQ), and stability at the Ewha Center for Vaccine Evaluation and Study, Seoul, Korea. We found that the specificity, reproducibility, and intermediate precision were within acceptance ranges (reproducibility, coefficient of variability [CV] ≤ 15%; intermediate precision, CV ≤ 20%) for all serotypes. Comparisons between the provisional assignments of calibration sera and the results from this laboratory showed a high correlation > 94% for all 7 serotypes, supporting the accuracy of the ELISA. The spiking recovery test also fell within an acceptable range. The quantification limit, calculated using the LLOQ, for each of the serotypes was 0.05-0.093 μg/mL. The freeze-thaw stability and the short-term temperature stability were also within an acceptable range. In conclusion, we showed good performance using the standardized WHO ELISA for the evaluation of serotype-specific anti-pneumococcal IgG antibodies; the WHO ELISA can evaluate the immune response against pneumococcal vaccines with consistency and accuracy.

Keyword

Streptococcus pneumoniae; Antibodies; Enzyme-Linked Immunosorbent Assay; Validation Studies

MeSH Terms

Antibodies*
Calibration
Enzyme-Linked Immunosorbent Assay
Global Health*
Humans*
Immunoglobulin G*
Immunoglobulins*
Korea
Pneumococcal Vaccines
Sensitivity and Specificity
Seoul
Serogroup
Streptococcus pneumoniae
World Health Organization*
Antibodies
Immunoglobulin G
Immunoglobulins
Pneumococcal Vaccines

Figure

• 

  Fig. 1 The specificity of anti-PnPS ELISA with the serum reference assay standard, 89-SF for 7 anti-PnPS types. Non-coating plate reference serum, 89-SF wells were coated with coating solution only. ELISA = enzyme-linked immunosorbent assay, PnPS = pneumococcal polysaccharide, OD = optical density.

• 

  Fig. 2 Short-term temperature stability of 5 calibration sera for 7 PnPS (acceptable range: within 100% ± 20%). For analysis of short term temperature, sera were left at room temperature for 1, 2, and 4 hours prior to analysis. PnPS = pneumococcal polysaccharide.

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