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. 1997 Jul;35(7):1751–1756. doi: 10.1128/jcm.35.7.1751-1756.1997

Interlaboratory agreement among results of human papillomavirus type 16 enzyme-linked immunosorbent assays.

H D Strickler 1, A Hildesheim 1, R P Viscidi 1, K V Shah 1, B Goebel 1, J Drummond 1, D Waters 1, Y Sun 1, N L Hubbert 1, S Wacholder 1, L A Brinton 1, C L Han 1, P C Nasca 1, R McClimens 1, K Turk 1, V Devairakkam 1, S Leitman 1, C Martin 1, J T Schiller 1
PMCID: PMC229834  PMID: 9196186

Abstract

Serological assays for measuring antibodies to human papillomavirus type 16 (HPV-16) virus-like particles (VLPs) have become important epidemiologic tools in recent years. However, the interlaboratory replicability of these assays has not been assessed. In this investigation, three laboratories tested a panel of specimens obtained from two different groups: 265 subjects in a vulvar cancer case-control study and 107 healthy volunteer blood donors. Each laboratory used an enzyme-linked immunosorbent assay (ELISA), but no attempt was made to standardize assay procedures among the three laboratories. The data showed good day-to-day intralaboratory replicability in laboratory 1 (correlation coefficient, > or = 0.88) and good intra-assay variability in laboratory 3 (correlation coefficient, > or = 0.93). Interlaboratory correlations, likewise, ranged between 0.61 and 0.80 in both case-control study subjects and healthy blood donors, indicating that ELISA optical density (OD) values between laboratories were linearly related regardless of the population. Kappa coefficients (kappa), based on each laboratory's categorical interpretation of its results (as positive or negative), showed good agreement (kappa, > 0.6) in case-control study subjects and moderate agreement (kappa, > or = 0.4) in blood donors, a population that had few strongly positive sera. When OD values near seropositive cutoffs were treated as indeterminates, there was little discordance between laboratories in either population. The data suggest that each laboratory measured the same humoral immune response and that their HPV-16 VLP ELISAs performed similarly (Pearson correlations). Interlaboratory differences, however, probably due to reagents and procedures, were considerably greater than intralaboratory day-to-day variability. Interlaboratory agreement in determining seropositivity (kappa) could be improved by sharing positive and negative serum controls and by treating marginal results as indeterminate. As part of continuing cooperation to improve interlaboratory agreement, we are preparing bulk serum control specimens to be shared and made available to interested researchers.

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Selected References

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  1. Dillner J., Wiklund F., Lenner P., Eklund C., Frederiksson-Shanazarian V., Schiller J. T., Hibma M., Hallmans G., Stendahl U. Antibodies against linear and conformational epitopes of human papillomavirus type 16 that independently associate with incident cervical cancer. Int J Cancer. 1995 Jan 27;60(3):377–382. doi: 10.1002/ijc.2910600318. [DOI] [PubMed] [Google Scholar]
  2. Galloway D. A. Papillomavirus capsids: a new approach to identify serological markers of HPV infection. J Natl Cancer Inst. 1994 Apr 6;86(7):474–475. doi: 10.1093/jnci/86.7.474. [DOI] [PubMed] [Google Scholar]
  3. Heino P., Eklund C., Fredriksson-Shanazarian V., Goldman S., Schiller J. T., Dillner J. Association of serum immunoglobulin G antibodies against human papillomavirus type 16 capsids with anal epidermoid carcinoma. J Natl Cancer Inst. 1995 Mar 15;87(6):437–440. doi: 10.1093/jnci/87.6.437. [DOI] [PubMed] [Google Scholar]
  4. Kirnbauer R., Booy F., Cheng N., Lowy D. R., Schiller J. T. Papillomavirus L1 major capsid protein self-assembles into virus-like particles that are highly immunogenic. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12180–12184. doi: 10.1073/pnas.89.24.12180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kirnbauer R., Hubbert N. L., Wheeler C. M., Becker T. M., Lowy D. R., Schiller J. T. A virus-like particle enzyme-linked immunosorbent assay detects serum antibodies in a majority of women infected with human papillomavirus type 16. J Natl Cancer Inst. 1994 Apr 6;86(7):494–499. doi: 10.1093/jnci/86.7.494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kirnbauer R., Taub J., Greenstone H., Roden R., Dürst M., Gissmann L., Lowy D. R., Schiller J. T. Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles. J Virol. 1993 Dec;67(12):6929–6936. doi: 10.1128/jvi.67.12.6929-6936.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lin L. I. A concordance correlation coefficient to evaluate reproducibility. Biometrics. 1989 Mar;45(1):255–268. [PubMed] [Google Scholar]
  8. Nonnenmacher B., Hubbert N. L., Kirnbauer R., Shah K. V., Muñoz N., Bosch F. X., de Sanjosé S., Viscidi R., Lowy D. R., Schiller J. T. Serologic response to human papillomavirus type 16 (HPV-16) virus-like particles in HPV-16 DNA-positive invasive cervical cancer and cervical intraepithelial neoplasia grade III patients and controls from Colombia and Spain. J Infect Dis. 1995 Jul;172(1):19–24. doi: 10.1093/infdis/172.1.19. [DOI] [PubMed] [Google Scholar]
  9. Palefsky J. M. Serologic detection of human papillomavirus-related anogenital disease: new opportunities and challenges. J Natl Cancer Inst. 1995 Mar 15;87(6):401–402. doi: 10.1093/jnci/87.6.401. [DOI] [PubMed] [Google Scholar]
  10. Strickler H. D., Schiffman M. H., Eklund C., Glass A. G., Scott D. R., Sherman M. E., Wacholder S., Kurman R. J., Manos M. M., Schiller J. T. Evidence for at least two distinct groups of humoral immune reactions to papillomavirus antigens in women with squamous intraepithelial lesions. Cancer Epidemiol Biomarkers Prev. 1997 Mar;6(3):183–188. [PubMed] [Google Scholar]
  11. Trimble C. L., Hildesheim A., Brinton L. A., Shah K. V., Kurman R. J. Heterogeneous etiology of squamous carcinoma of the vulva. Obstet Gynecol. 1996 Jan;87(1):59–64. doi: 10.1016/0029-7844(95)00351-7. [DOI] [PubMed] [Google Scholar]
  12. Viscidi R. P., Kotloff K. L., Clayman B., Russ K., Shapiro S., Shah K. V. Prevalence of antibodies to human papillomavirus (HPV) type 16 virus-like particles in relation to cervical HPV infection among college women. Clin Diagn Lab Immunol. 1997 Mar;4(2):122–126. doi: 10.1128/cdli.4.2.122-126.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wideroff L., Schiffman M. H., Nonnenmacher B., Hubbert N., Kirnbauer R., Greer C. E., Lowy D., Lorincz A. T., Manos M. M., Glass A. G. Evaluation of seroreactivity to human papillomavirus type 16 virus-like particles in an incident case-control study of cervical neoplasia. J Infect Dis. 1995 Dec;172(6):1425–1430. doi: 10.1093/infdis/172.6.1425. [DOI] [PubMed] [Google Scholar]
  14. Zweig M. H., Campbell G. Receiver-operating characteristic (ROC) plots: a fundamental evaluation tool in clinical medicine. Clin Chem. 1993 Apr;39(4):561–577. [PubMed] [Google Scholar]

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