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Related Experiment Videos

Between-method variation in human chorionic gonadotropin test results.

Laurence A Cole1, Jaime M Sutton, Trefor N Higgins

  • 1USA hCG Reference Service, Department of Obstetrics and Gynecology, University of New Mexico, Albuquerque, NM 87131, USA. larry@hcglab.com

Clinical Chemistry
|March 16, 2004
PubMed
Summary
This summary is machine-generated.

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Inter-assay variability in human chorionic gonadotropin (hCG) measurements is not due to hCG charge isoforms or structural variants. Differences in assay specificity and calibrator composition likely cause these variations in hCG testing.

Area of Science:

  • Clinical Chemistry
  • Immunoassay Development
  • Hormone Analysis

Background:

  • Commercial human chorionic gonadotropin (hCG) assays exhibit significant variability (1.4- to 2.3-fold) in results for sera and calibrators.
  • The specific contributions of calibrators, standards, hCG charge isoforms, and structural variants to this inter-assay variation remain unquantified.

Purpose of the Study:

  • To investigate the sources of inter-assay variation in human chorionic gonadotropin (hCG) measurements.
  • To determine the impact of hCG charge isoforms and structural variants on the performance of commercial hCG assays.

Main Methods:

  • Purified hCG was separated into four charge isoforms via isoelectric focusing.
  • Multiple hCG forms (pure hCG, hyperglycosylated hCG, hCG beta-subunit, nicked hCG, hCG beta-subunit core fragment) were analyzed across nine commercial hCG assays.

Related Experiment Videos

  • Immunoassay was used to determine the composition of hCG preparations and calibrators.

    • Main Results:

    • All four hCG charge isoforms showed parallel responses across the nine commercial assays.
    • Significant variations were observed in the detection of different hCG structural variants by the assays.
    • Commercial hCG preparations and calibrators displayed considerable variation in their content of hCG structural variants (34% to 100% intact hCG).

    Conclusions:

    • Inter-assay differences in hCG results are not explained by hCG charge isoforms or structural variants.
    • Assay specificity and the composition of calibrators are identified as the primary drivers of between-method variation in hCG testing.
    • Variations in calibrator composition and the detection of structural variants contribute to discrepancies in hCG measurements.