Early detection of the growth of Mycobacterium tuberculosis using magnetophoretic immunoassay in liquid culture.

Tuberculosis (TB) is an often neglected, epidemic disease that remains to be controlled by contemporary techniques of medicine and biotechnology. In this study, a nanoscale sensing system, referred to as magnetophoretic immunoassay (MPI) was designed to capture culture filtrate protein (CFP)-10 antigens effectively using two different types of nanoparticles (NPs). Two specific monoclonal antibodies against CFP-10 antigen were used, including gold NPs for signaling and magnetic particles for separation. These results were carefully compared with those obtained using the commercial mycobacteria growth indicator tube (MGIT) test via 2 sequential clinical tests (with ca. 260 clinical samples). The sensing linearity of MPI was shown in the range of pico- to micromoles and the detection limit was 0.3pM. MPI using clinical samples shows robust and reliable sensing while monitoring Mycobacterium tuberculosis (MTB) growth with monitoring time 3-10 days) comparable to that with the MGIT test. Furthermore, MPI distinguished false-positive samples from MGIT-positive samples, probably containing non-tuberculous mycobacteria. Thus, MPI shows promise in early TB diagnosis.

[1]  Madhukar Pai,et al.  Novel and improved technologies for tuberculosis diagnosis: progress and challenges. , 2009, Clinics in chest medicine.

[2]  C. Chiou,et al.  Rapid Identification of the Mycobacterium tuberculosis Complex by Combining the ESAT-6/CFP-10 Immunochromatographic Assay and Smear Morphology , 2010, Journal of Clinical Microbiology.

[3]  Eduardo Gotuzzo,et al.  Rapid molecular detection of tuberculosis and rifampin resistance. , 2010, The New England journal of medicine.

[4]  S. Shin,et al.  Identification and Diagnostic Utility of Serologic Reactive Antigens from Mycobacterium tuberculosis Sonic Extracts , 2009 .

[5]  Karen Steingart,et al.  New and improved tuberculosis diagnostics: evidence, policy, practice, and impact , 2010, Current opinion in pulmonary medicine.

[6]  S. Lawn,et al.  Point-of-care detection of lipoarabinomannan (LAM) in urine for diagnosis of HIV-associated tuberculosis: a state of the art review , 2012, BMC Infectious Diseases.

[7]  M. Hoelscher,et al.  Detection of mycobacterial lipoarabinomannan with an antigen-capture ELISA in unprocessed urine of Tanzanian patients with suspected tuberculosis. , 2005, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[8]  J. Andrews,et al.  HIV coinfection in multidrug- and extensively drug-resistant tuberculosis results in high early mortality. , 2010, American journal of respiratory and critical care medicine.

[9]  A. Buiting,et al.  Comparison of fluorescent BACTEC 9000 MB system, Septi-Chek AFB system, and Lowenstein-Jensen medium for detection of mycobacteria , 1996, Journal of clinical microbiology.

[10]  Jun Li,et al.  Size control of gold nanocrystals in citrate reduction: the third role of citrate. , 2007, Journal of the American Chemical Society.

[11]  Christopher Dye,et al.  The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. , 2003, Archives of internal medicine.

[12]  Simon S. Park,et al.  A plasmon-assisted fluoro-immunoassay using gold nanoparticle-decorated carbon nanotubes for monitoring the influenza virus. , 2015, Biosensors & bioelectronics.

[13]  Tae Jung Park,et al.  Rapid monitoring of CFP-10 during culture of Mycobacterium tuberculosis by using a magnetophoretic immunoassay , 2013 .

[14]  P. Andersen,et al.  Specific immune-based diagnosis of tuberculosis , 2000, The Lancet.

[15]  K. Koh,et al.  Green synthesis of phytochemical-stabilized Au nanoparticles under ambient conditions and their biocompatibility and antioxidative activity , 2011 .

[16]  H. Mollenkopf,et al.  Differential T cell responses to Mycobacterium tuberculosis ESAT6 in tuberculosis patients and healthy donors , 1998, European journal of immunology.

[17]  M. Pai,et al.  Diagnosing tuberculosis with urine lipoarabinomannan: systematic review and meta-analysis , 2011, European Respiratory Journal.

[18]  M. Pai,et al.  New Diagnostics for Latent and Active Tuberculosis: State of the Art and Future Prospects , 2008, Seminars in respiratory and critical care medicine.

[19]  M. Gennaro,et al.  Heterogeneous Antibody Responses in Tuberculosis , 1998, Infection and Immunity.

[20]  D. Fernig,et al.  Determination of size and concentration of gold nanoparticles from UV-vis spectra. , 2007, Analytical chemistry.

[21]  Jaebeom Lee,et al.  Clinical immunosensing of tuberculosis CFP-10 antigen in urine using interferometric optical fiber array , 2015 .

[22]  B. Hamasur,et al.  Diagnostic evaluation of urinary lipoarabinomannan at an Ethiopian tuberculosis centre. , 2001, Scandinavian journal of infectious diseases.

[23]  W. Rom,et al.  Current approaches to the diagnosis of active pulmonary tuberculosis. , 1994, American journal of respiratory and critical care medicine.

[24]  G. Alvarez-Uria Lung disease caused by nontuberculous mycobacteria , 2010, Current opinion in pulmonary medicine.

[25]  Yasuaki Maeda,et al.  Characterization and Catalytic Activity of Core−Shell Structured Gold/Palladium Bimetallic Nanoparticles Synthesized by the Sonochemical Method , 2000 .

[26]  D A Armbruster,et al.  Limit of detection (LQD)/limit of quantitation (LOQ): comparison of the empirical and the statistical methods exemplified with GC-MS assays of abused drugs. , 1994, Clinical chemistry.

[27]  G. Araj,et al.  Improved detection of mycobacterial antigens in clinical specimens by combined enzyme-linked immunosorbent assay. , 1993, Diagnostic microbiology and infectious disease.

[28]  S. Jeong,et al.  Evaluation of an Immunochromatographic Assay Kit for Rapid Identification of Mycobacterium tuberculosis Complex in Clinical Isolates , 2008, Journal of Clinical Microbiology.

[29]  C. Dye,et al.  Consensus statement. Global burden of tuberculosis: estimated incidence, prevalence, and mortality by country. WHO Global Surveillance and Monitoring Project. , 1999, JAMA.

[30]  Neus G Bastús,et al.  Kinetically controlled seeded growth synthesis of citrate-stabilized gold nanoparticles of up to 200 nm: size focusing versus Ostwald ripening. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[31]  S. Rüsch-Gerdes,et al.  Multicenter Evaluation of Fully Automated BACTEC Mycobacteria Growth Indicator Tube 960 System for Susceptibility Testing of Mycobacterium tuberculosis , 2002, Journal of Clinical Microbiology.