title: Use of multiepitope polyproteins in serodiagnosis of active tuberculosis.
abstract: Screening of genomic expression libraries from Mycobacterium tuberculosis with sera from tuberculosis (TB) patients or rabbit antiserum to M. tuberculosis led to the identification of novel antigens capable of detecting specific antibodies to M. tuberculosis. Three antigens, Mtb11 (also known as CFP-10), Mtb8, and Mtb48, were tested together with the previously reported 38-kDa protein, in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in TB patients. These four proteins were also produced as a genetically fused polyprotein, which was tested with two additional antigens, DPEP (also known as MPT32) and Mtb81. Sera from individuals with pulmonary and extrapulmonary TB, human immunodeficiency virus (HIV)-TB coinfections, and purified protein derivative (PPD)-positive and PPD-negative status with no evidence of disease were tested. In samples from HIV-negative individuals, the ELISA detected antibodies in >80% of smear-positive individuals and >60% smear-negative individuals, with a specificity of approximately 98%. For this group, smears detected 81.6% but a combination of smear and ELISA had a sensitivity of approximately 93%. The antigen combination detected a significant number of HIV-TB coinfections as well as antibodies in patients with extrapulmonary infections. Improved reactivity in the HIV-TB group was observed by including the antigen Mtb81 that was identified by proteomics. The data indicate that the use of multiple antigens, some of which are in a single polyprotein, can be used to facilitate the development of a highly sensitive test for M. tuberculosis antibody detection.
authors.affiliation: Corixa Corporation, Seattle, Washington 98104, USA. houghton@corixa.com
authors.forename: Raymond L
authors.initials: RL
authors.lastname: Houghton
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Lodes
authors.forename: Davin C
authors.initials: DC
authors.lastname: Dillon
authors.forename: Lisa D
authors.initials: LD
authors.lastname: Reynolds
authors.forename: Craig H
authors.initials: CH
authors.lastname: Day
authors.forename: Patricia D
authors.initials: PD
authors.lastname: McNeill
authors.forename: Ronald C
authors.initials: RC
authors.lastname: Hendrickson
authors.forename: Yasir A W
authors.initials: YA
authors.lastname: Skeiky
authors.forename: Diana P
authors.initials: DP
authors.lastname: Sampaio
authors.forename: Roberto
authors.initials: R
authors.lastname: Badaro
authors.forename: Konstantin P
authors.initials: KP
authors.lastname: Lyashchenko
authors.forename: Steven G
authors.initials: SG
authors.lastname: Reed
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D000939:Epitopes / Q000235:genetics; D015723:Gene Library; D015658:HIV Infections / Q000150:complications; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods / Q000592:standards; D014397:Tuberculosis, Pulmonary / Q000150:complications / Q000175:diagnosis*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000939:Epitopes; D011993:Recombinant Fusion Proteins