Korean J Pediatr.  2007 Feb;50(2):143-150. 10.3345/kjp.2007.50.2.143.

Validation of enzyme immunoassay for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae Type b capsular polysaccharide

• Kim KH
• Lim SY

• KMID: 2278717
• DOI: http://doi.org/10.3345/kjp.2007.50.2.143

Abstract

PURPOSE: This study was conducted to validate enzyme immunoassay (EIA) for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae type b (Hib) capsular polysaccharide. METHOD: We evaluated specificity, repeatability, intermediate precision, accuracy, lower limit of quantification (LLOQ), and stability to validate standardized EIA for the quantitative measurement of human anti-polyribosylribitol phosphate (PRP) IgG antibodies.
RESULTS
The results indicated that this EIA showed specificity to HbO-HA antigen and repeatability and intermediate precision were within acceptance criteria (repeatability: CV < or =15%, intermediate precision: CV < or =20%). The EIA-derived results from this laboratory were equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-PRP antibodies in the 28 sera. Spiking recovery result was within acceptance criteria (100+/-20%). The precision and accuracy of samples in LLOQ were from -14.7 to -4.7% in nominal values, which were within acceptance criteria (precision: CV < or =25%, accuracy: +/-25%). Freeze-thaw stability and short term temperature stability were within +/-20% of acceptance criteria.
CONCLUSIONS
The EIA which is performed at the Center for Vaccine Evaluation and Study Ewha Medical Research Institute, is an appropriate serologic assay which can be used for quantitation of anti-PRP IgG antibodies in human sera.

Keyword

Haemophilus influenzae type b; Antibodies; Enzyme immunoassay; Validation studies

MeSH Terms

Academies and Institutes
Antibodies*
Haemophilus influenzae type b*
Haemophilus influenzae*
Haemophilus*
Humans*
Immunoenzyme Techniques*
Immunoglobulin G*
Sensitivity and Specificity
Antibodies
Immunoglobulin G