All issues > Volume 50(2); 2007

Original Article

Korean J Pediatr. 2007;50(2):143-150. Published online February 15, 2007.

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

Kyung Hyo KH Kim¹, Soo Young SY Lim²

¹Department of Pediatrics, College of Medicine, Ewha Womans University
²Department of Microbiology, College of Medicine, Ewha Womans University

Correspondence Kyung Hyo KH Kim ,Email: kaykim@ewha.ac.kr

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 15%, intermediate precision: CV 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?0%). 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 25%, accuracy: ?5%). Freeze-thaw stability and short term temperature stability were within ?0% 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.

Keywords :Haemophilus influenzae type b , Antibodies , Enzyme immunoassay , Validation studies