Validation of enzyme immunoassay for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae Type b capsular polysaccharide.
10.3345/kjp.2007.50.2.143
- Author:
Kyung Hyo KIM
1
;
Soo Young LIM
Author Information
1. Department of Pediatrics, College of Medicine, Ewha Womans University, Korea. kaykim@ewha.ac.kr
- Publication Type:Validation Studies ; Original Article
- Keywords:
Haemophilus influenzae type b;
Antibodies;
Enzyme immunoassay;
Validation studies
- MeSH:
Academies and Institutes;
Antibodies*;
Haemophilus influenzae type b*;
Haemophilus influenzae*;
Haemophilus*;
Humans*;
Immunoenzyme Techniques*;
Immunoglobulin G*;
Sensitivity and Specificity
- From:Korean Journal of Pediatrics
2007;50(2):143-150
- CountryRepublic of Korea
- Language:Korean
-
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.
