Protein array identification of protein markers for serodiagnosis of Mycobacterium tuberculosis infection.
The lack of effective and accurate diagnostic tools contributes to the high prevalence of tuberculosis (TB) worldwide. The current serodiagnostics for TB are inadequate mainly due to lack of TB-specific antigens with highly accurate diagnosis. In the current study, we aimed to identify novel diagnostic antigens using glutathione S-transferase (GST)-fusion protein technique. We determined the reactivity of these recombinant proteins arrayed in solution and on GSH-immobilized microplates with TB patient sera. Of 409 TB proteins produced, ninety-two yielded seropositive reactions, fourteen including eight novel proteins showed strong immunoreactivity. Further, six were selected and constructed as a multiple-antigen combination set through analysis of various combinations. A comparative study of the multiple-antigen combination set and a commercially available kit revealed that the combination set showed 66.3% (95% CI 60.5-71.8) sensitivity, which was significantly higher than that of the commercial kit [31.6% (95% CI 26.3-37.3)]. The specificity of both methods was similar at 89.6% (95% CI 83.3-95.4) and 90.6% (95% CI 83.0-95.6), respectively. This study provides a set of novel diagnostic protein markers with great potential for the development of novel diagnostic tools for active TB.
Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.
Fangbin
F
Zhou
Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.
Xindong
X
Xu
Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.
Sijia
S
Wu
Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.
Xiaobing
X
Cui
Clinic and Research Center of Tuberculosis, Shanghai Key Lab of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China.
Lin
L
Fan
Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.|Department of Tropical Infectious Diseases, Second Military Medical University, Shanghai, China.
Weiqing
W
Pan
D000942:Antigens, Bacterial / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000276:immunology / Q000378:metabolism*; D015415:Biomarkers; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000276:immunology / Q000378:metabolism*; D016366:Open Reading Frames; D040081:Protein Array Analysis / Q000379:methods; D020543:Proteome; D040901:Proteomics / Q000379:methods; D011933:Reagent Kits, Diagnostic; D011993:Recombinant Fusion Proteins; D015203:Reproducibility of Results; D012698:Serologic Tests; D014376:Tuberculosis / Q000175:diagnosis* / Q000378:metabolism* / Q000382:microbiology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015415:Biomarkers; D020543:Proteome; D011933:Reagent Kits, Diagnostic; D011993:Recombinant Fusion Proteins