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. 1998 Dec 7;11(5):251–257. doi: 10.1002/(SICI)1098-2825(1997)11:5<251::AID-JCLA3>3.0.CO;2-A

Determination of IgE antibodies to the benzylpenicilloyl determinant: A comparison of the sensitivity and specificity of three radio allergo sorbent test methods

Juan J Garcia 1,✉, Miguel Blanca 1, Felisa Moreno 1, Jose M Vega 1, Cristobalina Mayorga 1, Javier Fernandez 2, Carlos Juarez 3,4, Antonino Romano 5, Enrique de Ramon 2
PMCID: PMC6760709  PMID: 9292392

Abstract

The quantitation of in vitro IgE antibodies to the benzylpenicilloyl determinant (BPO) is a useful tool for evaluating suspected penicillin allergic subjects. Although many different methods have been employed, few studies have compared their diagnostic specificity and sensitivity. In this study, the sensitivity and specificity of three different radio allergo sorbent test (RAST) methods for quantitating specific IgE antibodies to the BPO determinant were compared. Thirty positive control sera (serum samples from penicillin allergic subjects with a positive clinical history and a positive penicillin skin test) and 30 negative control sera (sera from subjects with no history of penicillin allergy and negative skin tests) were tested for BPO‐specific IgE antibodies by RAST using three different conjugates coupled to the solid phase: benzylpenicillin conjugated to polylysine (BPO‐PLL), benzylpenicillin conjugated to human serum albumin (BPO‐HSA), and benzylpenicillin conjugated to an aminospacer (BPO‐SP). Receiver operator control curves (ROC analysis) were carried out by determining different cut‐off points between positive and negative values. Contingence tables were constructed and sensitivity, specificity, negative predictive values (PV–), and positive predictive values (PV+) were calculated. Pearson correlation coefficients (r) and intraclass correlation coefficients (ICC) were determined and the differences between methods were compared by χ2 analysis. Analysis of the areas defined by the ROC curves showed statistical differences among the three methods. When cut‐off points for optimal sensitivity and specificity were chosen, the BPO‐HSA assay was less sensitive and less specific and had a lower PV– and PV+ than the BPO‐PLL and BPO‐SP assays. Assessment of r and ICC indicated that the correlation was very high, but the concordance between the PLL and SP methods was higher than between the PLL and HSA or SP and HSA methods. We conclude that for quantitating IgE antibodies by RAST to the BPO determinant, BPO‐SP or BPO‐PLL conjugates offer advantages in sensitivity and specificity compared with BPO‐HSA. These results support and extend previous in vitro studies by our group and highlight the importance of the carrier for RAST assays. J. Clin. Lab. Anal. 11:251–257, 1997. © 1997 Wiley‐Liss, Inc.

Keywords: benzylpenicilloyl, IgE antibodies, polylysine, human serum albumin, spacer

References

  • 1. Batchelor FR, Dewdney JM, Gazzard D: Penicillin allergy: The formation of the penicilloyl determinant. Nature 6: 362–364, 1965. [DOI] [PubMed] [Google Scholar]
  • 2. Levine BB: Immunochemical involved in penicillin hypersensitivity in experimental animals and in human beings. Fed Proc 24: 45–50, 1965. [PubMed] [Google Scholar]
  • 3. De Week AL, Blum G: Recent clinical and immunochemical aspects of penicillin allergy. Int Arch Allergy Appl Immunol 27: 221–229, 1965. [DOI] [PubMed] [Google Scholar]
  • 4. Parker CW, Shapiro J, Kern J, Eisen HN: Hypersensitivity to penicillenic acid derivatives in human beings with penicillin allergy. J Exp Med 115: 821–828, 1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. Dewdney JM: Immunology of the antibiotics In Antigens 5. Sela M, ed. Academic Press, New York, 1977, p 73–245. [Google Scholar]
  • 6. Kraft D, Berglund A, Rumpold H, Roth A, Ebner H: Radioallergosorbent test with conjugates specific for “minor” haptenic determinants in the diagnosis of IgE‐mediated penicillin allergy in man. Clin Allergy 11: 579–587, 1981. [DOI] [PubMed] [Google Scholar]
  • 7. Edwards RG, Spackman DA, Dewdney JM: Development and use of three new radioallergosorbent tests in the diagnosis of penicillin allergy. Int Arch Allergy Appl Immunol 68: 352–357, 1982. [DOI] [PubMed] [Google Scholar]
  • 8. Adkinson NF, Thompson WL, Maddrey WC, Lichtenstein LM: Routine use of penicillin skin testing on an inpatient service. N Engl J Med 285: 22–24, 1971. [DOI] [PubMed] [Google Scholar]
  • 9. Juhlin L, Wide L: Detection of penicillin allergy by the radioallergosorbent test. Lancet 2: 261, 1979. [DOI] [PubMed] [Google Scholar]
  • 10. Jarisch R, Roth A, Boltz A, Sandor I: Diagnosis of penicillin allergy by means of Phadebas RAST penicilloyl G and V and skin tests. Clin Allergy 11: 155–160, 1981. [DOI] [PubMed] [Google Scholar]
  • 11. Christie G, Coleman W, Newby S, McDiarmaid‐Gordon A, Hampson JP, Breckenridge AM, Park BK: A survey of the prevalence of penicillin‐specific IgG, IgM and IgE antibodies detected by ELISA and defined by hapten inhibition, in patients with suspected penicillin allergy and in healthy volunteers. Br J Clin Pharmacol 25: 381–386, 1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. Kraft D, Wilson DV, Dervey ME: Penicillin allergy studies by modified red‐cell‐linked antigen‐antiglobulin reaction. Int Arch Allergy Appl Immunol 52: 248–256, 1976. [DOI] [PubMed] [Google Scholar]
  • 13. Ohno Y, Stahmann M: A new detection method for penicillin antibodies and hypersensitivity in humans using penicillin coupled to polyacrylamide particles. Immunochemistry 9: 1087–1093, 1972. [DOI] [PubMed] [Google Scholar]
  • 14. de Haan P, Kalsbeek GL: Induction of benzylpenicilloyl specific antibodies, including IgE, by long‐term administration of benzylpenicillin. Clin Allergy 13: 563–569, 1983. [DOI] [PubMed] [Google Scholar]
  • 15. Harle DG, Baldo BA: Drugs as allergens: An immunoassay for detecting IgE antibodies to cephalosporins. Int Arch Allergy Appl Immunol 92: 439–444, 1990. [DOI] [PubMed] [Google Scholar]
  • 16. Daxun Z, Adriansson J, Martin A, Malmberg E, Ahlatedt S: Diagnostic capabilities of penicillin‐based structures as tools in the in vitro diagnosis of penicillin allergy. J Invest Allergol Clin Immunol 2: 240–246, 1992. [PubMed] [Google Scholar]
  • 17. Klein GF, Stanzl U, Fritsch PO, Varga JM: Enzyme‐linked immunosorbent assay for detection of antibodies to the unmodified beta‐lactam ring. Allergy 48: 151–157, 1993. [DOI] [PubMed] [Google Scholar]
  • 18. Wide L, Juhlin L: Detection of penicillin allergy of the immediate type by radioimmunoassay of reagins (IgE) to penicilloyl conjugates. Clin Allergy 1: 171–175, 1971. [DOI] [PubMed] [Google Scholar]
  • 19. Urena V, Delgado RG, Daroca, Lahoz C: Polystyrene tube radioimmunoabsorbent assay for IgE anti‐penicillin antibodies. Lancet 1: 1210–1211, 1977. [DOI] [PubMed] [Google Scholar]
  • 20. Blanca M, Mayorga C, Pérez E, Suau R, Juárez C, Vega JM, Carmona MJ, Perez‐Estrada M, García J: Determination of IgE antibodies to the benzyl penicilloyl determinant. A comparison between poly‐L‐lysine and human serum albumin as carriers. J Immunol Methods 153: 99–105, 1992. [DOI] [PubMed] [Google Scholar]
  • 21. de Haan P, Kalsbeeck GL: The humoral immune response to some ben‐zylpenicillin preparations. Allergy 38: 319–324, 1983. [DOI] [PubMed] [Google Scholar]
  • 22. Isetta AM, Minghetti A, Radojkovic J, Vasile C, Celada F: Affinity and specificity of penicillin‐antibody interaction determined by an enzyme (E. coli β‐galactosidase) immunoassay. Eur J Immunol 6: 737–742, 1976. [DOI] [PubMed] [Google Scholar]
  • 23. Botello A, Moneo I, Cuevas M, Alvarez E: Detección de anticuerpos anti‐penicilina: correlación entre la técnica de RAST y el enzimoinmunoanálisis a doble paso. Allergol Immunopathol (Suppl VII): 297, 1980.
  • 24. Mamas S, Dray F: Viroimmunoenzymoassay for detection of anti‐penicilloyl antibodies and penicilloyl residues: Comparison of results obtained by radio‐, viro‐ and enzymoimmunoassay. Ann Immunol Inst Pasteur 30: 407–418, 1979. [PubMed] [Google Scholar]
  • 25. Caneva E, Di Genaro P, Farina F, Orlandi M, Rindone B: Synthesis and characterization of a penicillin‐poly(L‐lysine) which recognizes human IgE anti‐penicillin antibodies. Bioconjugate Chem 4: 309–314, 1993. [DOI] [PubMed] [Google Scholar]
  • 26. Moreno F, Blanca M, Mayorga C, Terrados S, Moya MC, Pérez E, Suau R: Studies of the specificities of IgE antibodies found in sera from subjects with allergic reactions to penicillins. Int Arch Allergy Appl Immunol 108: 74–81, 1995. [DOI] [PubMed] [Google Scholar]
  • 27. Mayorga C, Obispo T, Jimeno L, Blanca M, Moscoso J, Carreira J, Garcia JJ, Juárez C: Epitope mapping of beta lactams antibiotics with the use of monoclonal antibodies. Toxicology 97: 225–234, 1995. [DOI] [PubMed] [Google Scholar]
  • 28. Blanca M, Vega JM, García J, Carmona MJ, Terrados S, Avila MJ, Miranda A, Juárez C: Allergy to amoxicillin with good tolerance to other penicillins. Study of the incidence in patients allergic to beta lactams. Clin Exp Allergy 20: 475–481, 1991. [DOI] [PubMed] [Google Scholar]
  • 29. Blanca M, Moreno F, Mayorga C, García J, Fernández M, Pérez E, Juárez C, Suau R: The nature of the carrier in the RAST assay influences the capacity for detecting IgE antibodies to penicillins. J Clin Immunoassay 17: 1–5, 1994. [Google Scholar]
  • 30. Sackett DL: Basic research. Clinical research. Clinical epidemiology and general internal medicine. J Gen Intern Med 2: 40–47, 1987. [DOI] [PubMed] [Google Scholar]
  • 31. Bartko: The intraclass correlation coefficient as a measure of reliability. Psychol Rep 19: 3–11, 1966. [DOI] [PubMed] [Google Scholar]
  • 32. Weiss ME, Adkinson NF: Immediate hypersensitivity reactions to penicillin and related antibiotics. Clin Allergy 18: 515–540, 1988. [DOI] [PubMed] [Google Scholar]
  • 33. Levine BB, Zolov DM: Prediction of penicillin allergy by immunological tests. J Allergy 43: 231–244, 1969. [DOI] [PubMed] [Google Scholar]
  • 34. Gadde J, Spence M, Wheeler B, Adkinson F: Clinical experience with penicillin skin testing in a large inner‐city STD clinic. JAMA 20: 2456–2463, 1993. [PubMed] [Google Scholar]
  • 35. Blanca M, Fernández J, Miranda A, Terrados S, Torres MJ, Vega JM, Avila MJ, Pérez E, García J, Suau R: Cross‐reactivities between penicillin and cephalosporins. Clinical and Immunological Studies. J Allergy Clin Immunol 83: 381–385, 1989. [DOI] [PubMed] [Google Scholar]
  • 36. Lew AM: The effect of epitope density and antibody affinity on the ELISA as analysed by monoclonal antibodies. J Immunol Methods 72: 171–176, 1984. [DOI] [PubMed] [Google Scholar]
  • 37. Lee D, Dewdney JM, Edwards RG: The influence of hapten density on the assay of penicilloylated proteins in fluids. J Immunol Methods 84: 235–243, 1985. [DOI] [PubMed] [Google Scholar]
  • 38. Lafaye P, Lapresle C: Fixation of penicilloyl groups to albumin and appearance of anti‐penicilloyl antibodies in penicillin‐treated patients. J Clin Invest 82: 7–12, 1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39. Ressler C, Mendelson LM: Skin test for diagnosis of penicillin allergy‐current status. Ann Allergy 59: 167–170, 1987. [PubMed] [Google Scholar]
  • 40. Parker CW, Thiel JA: Studies in human penicillin allergy: A comparison of various penicilloyl‐polylysines. J Lab Clin Med 3: 482–491, 1963. [PubMed] [Google Scholar]

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