Open AccessCloning and Expression of Immunoreactive Antigens from Mycobacterium tuberculosis
Author(s) -
Renee Lay Hong Lim,
Li Tan,
W. Lau,
Maxey Ching Ming,
Hyun Cheol Chung,
Roseanne Dunn,
Heng Phon Too,
Lily Chan
Publication year - 2000
Publication title -
clinical and diagnostic laboratory immunology
Language(s) - English
Resource type - Journals
eISSN - 1098-6588
pISSN - 1071-412X
DOI - 10.1128/cdli.7.4.600-606.2000
Subject(s) - recombinant dna , mycobacterium tuberculosis , antigen , tuberculosis , biology , antibody , microbiology and biotechnology , virology , cloning (programming) , expression vector , gene , immunology , genetics , medicine , pathology , computer science , programming language
Four immunoreactive proteins, B.4, B.6, B.10, and B.M, with molecular weights ranging from 16,000 to 58,000, were observed from immunoblots ofMycobacterium tuberculosis total lysates screened with sera from individuals with active tuberculosis. These proteins were identified from microsequence analyses, and genes of proteins with the highest homology were PCR amplified and cloned into the pQE30 vector for expression studies. In addition, a 37.5-kDa protein, designated C17, was identified from a phage expression library ofM. tuberculosis genomic DNA. Preliminary immunoblot assays indicated that these five resultant recombinant proteins could detect antibodies in individuals with active pulmonary and extrapulmonary tuberculosis. The overall ranges of sensitivities, specificities, positive predictive values, and negative predictive values for the recombinant antigens were 20 to 58, 88 to 100, 69 to 100, and 56 to 71%, respectively. The B.6 antigen showed preferential reactivity to antibodies in pulmonary compared to nonpulmonary tuberculosis serum specimens. All of these recombinant antigens demonstrated potential for serodiagnosis of tuberculosis.