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Antibody-free detection of Mycobacterium tuberculosis antigen using customized nanotraps.

ABSTRACT: Rapid screening and diagnosis of tuberculosis disease (TB) is still challenging and critically needed for global TB control efforts. In this study, we present a rapid and streamlined technology, using precisely engineered silica nanopore thin films, which are optimized for pore size, structure, capillary force, and film thickness, to isolate Mycobacterium tuberculosis (MTB) antigens in laboratory and clinical samples for rapid TB screening. This technology, referred to here as on-chip fractionation, is integrated with high-throughput matrix-assisted laser desorption/ionization time-of flight mass spectrometry to screen and identify fragments of the MTB antigen, CFP-10, from complex biological samples, without use of immunoaffinity agents. With the use of this comprehensive approach, we were able to clearly distinguish a clinical isolate of MTB from a nonTB species of the genus Mycobacterium avium grown in liquid culture media. This assay can reach a detection limit of 10 fmol and an isolation rate of 90% for the antigen CFP-10. Our strategy has significant potential to fill the conceptual and technical gaps in rapid diagnosis of active TB disease.

SUBMITTER: Wu HJ

PROVIDER: S-EPMC3983014 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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Antibody-free detection of Mycobacterium tuberculosis antigen using customized nanotraps.

Wu Hung-Jen HJ Li Yaojun Y Fan Jia J Deng Zaian Z Hu Zhao Z Liu Xuewu X Graviss Edward A EA Ferrari Mauro M Ma Xin X Hu Ye Y

Analytical chemistry 20140205 4

Rapid screening and diagnosis of tuberculosis disease (TB) is still challenging and critically needed for global TB control efforts. In this study, we present a rapid and streamlined technology, using precisely engineered silica nanopore thin films, which are optimized for pore size, structure, capillary force, and film thickness, to isolate Mycobacterium tuberculosis (MTB) antigens in laboratory and clinical samples for rapid TB screening. This technology, referred to here as on-chip fractionat ...[more]

PMID: 24446580

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Project description:Clinical and laboratory diagnosis of Active Tuberculosis (ATB) and latent Mycobacterium Tuberculosis (M. tuberculosis) infections (LTBI) among people living with HIV/AIDS (PLWHA) presents formidable challenges. In the past, WHO issued an advisory against the use of existing TB sero-diagnostics. Emerging evidence, however, points to a precision of TB sero-diagnostics based on secretory rather than structural M. tuberculosis antigens. We hypothesized that secretory levels of M. tuberculosis thymidylate kinase (TMKmt) can Designate ATBI from LTBI and no TB (NTB). Here, we report in-house validation studies of levels of TMKmt antigen (Ag) and host specific TMKmt antibody (Ab) amongst HIV +ve and HIV -ve participants. Direct TMKmt Ag and host specific IgG Ab detection EIAs were conducted on broadly consented, stored serum (N=281[Ag] vs. 214 [Ab] respective) samples stratified as either HIV +ve or HIV-ve ATB relative to LTBI and No TB. On one hand, UG-peptide 1 and its PAb-based EIAs accurately diagnosed ATB relative to LTBI and NTB among HIV +ve subjects {irrespectively: (a) Ag detection ATB=OD>0.490; 95% CI: 0.7446 to 0.8715 vs. LTBI=OD<0.490; 95% CI 0.4325 to 0.4829 vs. NTB=OD<0.26; 95% CI 0.1675 to 0.2567 and (b) TMKmt specific IgG detection ATB=OD>1.00; 95% CI 1.170 to 1.528 [HIV +ve] and 2.044 to 2.978 [HIV -ve] respectively vs. LTBI=OD<1.00; 95% CI 0.2690 to 0.6396 vs. NTB=OD<; 95% CI 0.1527 to 0.8751}. HIV -ve ATB presented with Ag levels greater than NTB and less than LTBI (i.e. ATB -ve=<0.490 ODs>0.26), but displayed better ant-TMKmt IgG responses (OD>2.00; 95% CI 2.044 to 2.978) relative to HIV +ve ATB (OD<1.600; 95% CI 1.170 to 1.528); suggesting a better control of M. tuberculosis-septicemia. On the other hand, UG-peptide 2 and its PAb-based EIAs did not demonstrate ATB diagnostic potential regardless of HIV sero-status, except towards designating NTB. TMKmt Ab and Ag detecting EIAs based on UG-peptide 1 and its derivative PAb can accurately demarcate ATB from LTBI and NTB among HIV +ve subjects.

Dataset Information

Antibody-free detection of Mycobacterium tuberculosis antigen using customized nanotraps.

Publications

Antibody-free detection of Mycobacterium tuberculosis antigen using customized nanotraps.

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