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Clinical and Diagnostic Laboratory Immunology, November 2005, p. 1311-1316, Vol. 12, No. 11
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.11.1311-1316.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Selected RD1 Peptides for Active Tuberculosis Diagnosis: Comparison of a Gamma Interferon Whole-Blood Enzyme-Linked Immunosorbent Assay and an Enzyme-Linked Immunospot Assay

Delia Goletti,1,2* Donatella Vincenti,2 Stefania Carrara,2 Ornella Butera,2 Federica Bizzoni,2 Giuliana Bernardini,3 Massimo Amicosante,4 and Enrico Girardi5

Second Division, Health Department of the National Institute for Infectious Diseases (INMI) Lazzaro Spallanzani,1 Translational Research Unit, INMI,2 Department of Internal Medicine, University of Tor Vergata,4 Epidemiology Department, INMI, Rome,5 Division of Infectious Diseases, Ospedale Belcolle, Viterbo, Italy3

Received 23 June 2005/ Returned for modification 2 August 2005/ Accepted 25 August 2005

We recently set up a gamma interferon (IFN-{gamma}) enzyme-linked immunospot assay (ELISPOT), using selected early secreted antigenic target 6 (ESAT-6) peptides, that appears specific for active tuberculosis (A-TB). However, ELISPOT is difficult to automate. Thus, the objective of this study was to determine if the same selected peptides may be used in a technique more suitable for routine work in clinical laboratories, such as whole-blood enzyme-linked immunosorbent assay (WBE). For this purpose, 27 patients with A-TB and 41 control patients were enrolled. Our WBE, using the already described selected peptides from ESAT-6 plus three new ones from culture filtrate protein 10, was performed, and data were compared with those obtained by ELISPOT. Using our selected peptides, IFN-{gamma} production, evaluated by both WBE and ELISPOT, was significantly higher in patients with A-TB than in controls (P < 0.0001). Statistical analysis showed a good correlation between the results obtained by WBE and ELISPOT (r = 0.80, P < 0.001). To substantiate our data, we compared our WBE results with those obtained by QuantiFERON-TB Gold, a whole-blood assay based on region of difference 1 (RD1) overlapping peptides approved for TB infection diagnosis. We observed a slightly higher sensitivity with QuantiFERON-TB Gold than with our WBE (89% versus 81%); however, our test provided a better specificity result (90% versus 68%). In conclusion, results obtained by WBE based on selected RD1 peptides significantly correlate with those generated by ELISPOT. Moreover, our assay appears more specific for A-TB diagnosis than QuantiFERON-TB Gold, and thus it may represent a complementary tool for A-TB diagnosis for routine use in clinical laboratories.

* Corresponding author. Mailing address: Laboratorio di collegamento tra ricerca di base e clinica, Padiglione Del Vecchio, Istituto Nazionale per le Malattie Infettive Lazzaro Spallanzani, I.R.C.C.S., Roma 00149, Italy. Phone: 39-06-55170-954. Fax: 39-06-55170-904. E-mail: d.goletti{at}tiscali.it.

Clinical and Diagnostic Laboratory Immunology, November 2005, p. 1311-1316, Vol. 12, No. 11
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.11.1311-1316.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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