Antibody-free detection of Mycobacterium tuberculosis antigen using customized nanotraps.
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.
Department of Nanomedicine, Houston Methodist Research Institute , 6670 Bertner Avenue R8-213, Houston, TX 77030, United States.
Hung-Jen
HJ
Wu
Yaojun
Y
Li
Jia
J
Fan
Zaian
Z
Deng
Zhao
Z
Hu
Xuewu
X
Liu
Edward A
EA
Graviss
Mauro
M
Ferrari
Xin
X
Ma
Ye
Y
Hu
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000032:analysis* / Q000235:genetics; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods*; D056951:Photoelectron Spectroscopy / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000942:Antigens, Bacterial; C072681:Mycobacterium tuberculosis antigens