ESAT-6/CFP-10 fusion protein and peptides for optimal diagnosis of Mycobacterium tuberculosis infection by ex vivo enzyme-linked immunospot assay in The Gambia

被引:54
作者
Hill, PC
Jackson-Sillah, D
Fox, A
Franken, KLMC
Lugos, MD
Jeffries, DJ
Donkor, SA
Hammond, AS
Adegbola, RA
Ottenhoff, THM
Klein, MR
Brookes, RH
机构
[1] MRC Unit, Bacterial Dis Program, TB Div, Banjul, Gambia
[2] Leiden Univ, Med Ctr, Dept Immunohematol & Blood Transfus, Leiden, Netherlands
基金
英国医学研究理事会;
关键词
D O I
10.1128/JCM.43.5.2070-2074.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Overlapping peptides of Mycobacterium tuberculosis antigens ESAT-6 and CFP-10 offer increased specificity over the purified protein derivative skin test when they were used in an ex vivo enzyme-linked immunospot (ELISPOT) assay for gamma interferon detection for the diagnosis of M. tuberculosis infection from recent exposure. We assessed whether equivalent results could be obtained for a fusion protein of the two antigens and whether a combined readout would offer increased sensitivity in The Gambia. We studied the ELISPOT assay results for 488 household contacts of 88 sputum smear-positive tuberculosis (TB) cases. The proportions of subjects positive by each test and by the tests combined were assessed across an exposure gradient, defined according to sleeping proximity to a TB case. Eighty-eight (18%) subjects were positive for CFP-10 peptides, 148 (30%) were positive for ESAT-6 peptides, 161 (33%) were positive for both peptides, and 168 (34%) were positive for the fusion protein; 188 (39%) subjects had either a positive result for a peptide or a positive result for the fusion protein. There was reasonable agreement between the peptide and the protein results (kappa statistic = 0.78) and no significant discordance (P = 0.38). There was a strong correlation between the fusion protein and combined peptide spot counts (r = 0.9), and responses to the peptide and the proteins all increased significantly according to M. tuberculosis exposure. The proportion of subjects positive for either the pool of peptides or the fusion protein offered maximum sensitivity, being significantly higher than the proportion of subjects positive for ESAT-6 peptides alone (P = 0.007). A fusion protein of ESAT-6 and CFP-10 is equivalent to overlapping peptides for the diagnosis of latent M. tuberculosis infection. Use of a combination of peptides and fusion protein offers improved sensitivity.
引用
收藏
页码:2070 / 2074
页数:5
相关论文
共 16 条
[1]   Tuberculin skin testing compared with T-cell responses to Mycobacterium tuberculosis-specific and nonspecific antigens for detection of latent infection in persons with recent tuberculosis contact [J].
Arend, SM ;
Engelhard, ACF ;
Groot, G ;
de Boer, K ;
Andersen, P ;
Ottenhoff, THW ;
van Dissel, JT .
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 2001, 8 (06) :1089-1096
[2]   Detection of active tuberculosis infection by T cell responses to early-secreted antigenic target 6-kDa protein and culture filtrate protein 10 [J].
Arend, SM ;
Andersen, P ;
van Meijgaarden, KE ;
Skjot, RLV ;
Subronto, YW ;
van Dissel, JT ;
Ottenhoff, THM .
JOURNAL OF INFECTIOUS DISEASES, 2000, 181 (05) :1850-1854
[3]   Diagnosing latent tuberculosis infection - Turning glitter to gold [J].
Barnes, PF .
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 2004, 170 (01) :5-6
[4]   A Mycobacterium tuberculosis operon encoding ESAT-6 and a novel low-molecular-mass culture filtrate protein (CFP-10) [J].
Berthet, FX ;
Rasmussen, PB ;
Rosenkrands, I ;
Andersen, P ;
Gicquel, B .
MICROBIOLOGY-UK, 1998, 144 :3195-3203
[5]   Rapid detection of active and latent tuberculosis infection in HIV-positive individuals by enumeration of Mycobacterium tuberculosis-specific T cells [J].
Chapman, ALN ;
Munkanta, M ;
Wilkinson, KA ;
Pathan, AA ;
Ewer, K ;
Ayles, H ;
Reece, WH ;
Mwinga, A ;
Godfrey-Faussett, P ;
Lalvani, A .
AIDS, 2002, 16 (17) :2285-2293
[6]   Purification of His-tagged proteins by immobilized chelate affinity chromatography: The benefits from the use of organic solvent [J].
Franken, KLMC ;
Hiemstra, HS ;
van Meijgaarden, KE ;
Subronto, Y ;
den Hartigh, J ;
Ottenhoff, THM ;
Drijfhout, JW .
PROTEIN EXPRESSION AND PURIFICATION, 2000, 18 (01) :95-99
[7]   Immune recognition of a human renal cancer antigen through post-translational protein splicing [J].
Hanada, K ;
Yewdell, JW ;
Yang, JC .
NATURE, 2004, 427 (6971) :252-256
[8]  
Heifets Leonid B., 1994, P85
[9]   Large-scale evaluation of enzyme-linked immunospot assay and skin test for diagnosis of Mycobacterium tuberculosis infection against a gradient of exposure in The Gambia [J].
Hill, PC ;
Brookes, RH ;
Fox, A ;
Fielding, K ;
Jeffries, DJ ;
Jackson-Sillah, D ;
Lugos, MD ;
Owiafe, PK ;
Donkor, SA ;
Hammond, AS ;
Otu, JK ;
Corrah, T ;
Adegbola, RA ;
McAdam, KPWJ .
CLINICAL INFECTIOUS DISEASES, 2004, 38 (07) :966-973
[10]   Rapid effector function in CD8(+) memory T cells [J].
Lalvani, A ;
Brookes, R ;
Hambleton, S ;
Britton, WJ ;
Hill, AVS ;
McMichael, AJ .
JOURNAL OF EXPERIMENTAL MEDICINE, 1997, 186 (06) :859-865