ORFeome-based identification of biomarkers for serodiagnosis of Mycobacterium tuberculosis latent infection

Abstract

BackgroundThe challenges posed by Mycobacterium tuberculosis infection require the gradual removal of the pool of latent tuberculosis infection (LTBI). The current cell-immune-based diagnostic tests used to identify LTBI individuals have several irreversible drawbacks. In the present study, we attempted to identify novel diagnostic antigens for LTBI.MethodsA high-throughput glutathione S-transferase (GST)-fusion technology was used to express over 409 TB proteins and sera from LTBI and healthy individuals was used to interrogate these GST-TB fusion proteins.ResultsOf 409 TB proteins, sixty-three reacted seropositive and defined the immuno-ORFeome of latent M. tuberculosis. Within the immuno-ORFeome, the rare targets were predominantly latency-associated proteins and secreted proteins, while the preferentially recognized antigens tended to be transmembrane proteins. Six of novel highly-reactive antigens had the potential to distinguish LTBI from active TB and healthy individuals. A multiple-antigen combination set was selected through analysis of various combinations. A panel of 94 archived serum samples was used to validate the diagnostic performance of the multiple-antigen combination set, which had sensitivity of 66.1% (95% CI 52.9, 77.4) and specificity of 87.5% (95% CI 70.1, 95.1).ConclusionThese results provide experimental evidence of the immunogenicity of novel TB proteins that are suitable for the development of serodiagnostic tools for LTBI.Electronic supplementary materialThe online version of this article (10.1186/s12879-017-2910-y) contains supplementary material, which is available to authorized users.

Immunoreactive Proteins of Dormant Mycobacterium tuberculosis Cells

“…However, the immunogenicity of proteins even in this group varied significantly among patients with TB. A similar large variability in immunogenicity was described in a recent publication on TB serodiagnosis in 755 patients from several countries with a panel of 22 Mtb antigens preselected from the total proteome in the form of recombinant proteins [14], as well as in a work on the identification of biomarkers for the serodiagnosis of latent TB [22].…”

Shotgun proteomic profiling of dormant, ‘non-culturable’Mycobacterium tuberculosis

“…6). Despite the serodiagnosis of latent TB being very obscure, the work of Zhou et al (32), where serodiagnostic analysis was carried out using the sera of patients with confirmed latent TB at the proteome level, is of particular interest. Among 62 seropositive TB proteins reported by Zhou et al, 12 proteins out of 40 were found in the current study.…”

“…The shortened list of proteins was further compared with previously published datasets with annotated Mtb proteins with antigenic properties: i) a list of 181 proteins detected exclusively in the sera of TB-positive patients from a study on the dynamic antibody response to the proteome (31); ii) a list of 62 proteins resulting from serodiagnosis of latent Mtb infection (32); iii) a list of 22 proteins resulting from the analysis of TB-specific IgG antibody profiles, detectable on two different platforms (Luminex and MBio) (33).…”

Assessment of a Fusion Polyprotein Based on HspX and Other Eight Antigen Segments for Tuberculosis Serodiagnosis

“…However, none of any single antigen had enough high sensitivity. 15,16 In this study, we aimed to combine nine antigens (or segments) with un-conserved dominant B-cell epitopes to be a fusion polyprotein, which may improve the whole sensitivity and specificity compared with individual antigen and other combination forms. and E. coli were used to pre-adsorb the serum samples to block antibodies against bacteriaantigens and reduce non-specific reactions.In details,Bacteria were cultured at 100 mL volume overnight at 37℃ and then collected after centrifugation at 13,000 rpm at 4°C for 10 min.…”