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Rapid detection of Mycobacterium tuberculosis biomarkers in a sandwich immunoassay format using a waveguide-based optical biosensor

 Harshini Mukundan  ;  Sandeep Kumar  ;  Dominique N. Price  ;  Sonja M. Ray  ;  Ye-Jin Lee  ;  Seonyeong Min  ;  Seokyong Eum  ;  Jessica Kubicek-Sutherland  ;  Jesse M. Resnick  ;  W. Kevin Grace  ;  Aaron S. Anderson  ;  Soo Hee Hwang  ;  Sang Nae Cho  ;  Laura E. Via  ;  Clifton Barry III  ;  Ramamurthy Sakamuri  ;  Basil I. Swanson 
 TUBERCULOSIS, Vol.92(5) : 407-416, 2012 
Journal Title
Issue Date
Acyltransferases/metabolism* ; Antigens, Bacterial/metabolism* ; Bacterial Proteins/metabolism* ; Biomarkers/metabolism ; Biosensing Techniques* ; Coinfection ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Seropositivity/epidemiology ; HIV Seropositivity/metabolism* ; Humans ; Immunoassay ; Lipopolysaccharides/metabolism* ; Male ; Mycobacterium tuberculosis/isolation & purification* ; Mycobacterium tuberculosis/pathogenicity ; Reagent Kits, Diagnostic ; Reproducibility of Results ; Sensitivity and Specificity ; Tuberculosis, Pulmonary/diagnosis ; Tuberculosis, Pulmonary/epidemiology ; Tuberculosis, Pulmonary/metabolism*
Tuberculosis ; Diagnosis ; Lipoarabinomannan ; Early secretory antigenic target 6 ; Antigen 85 complex ; Waveguide-based optical biosensor
Early diagnosis of active tuberculosis (TB) remains an elusive challenge, especially in individuals with disseminated TB and HIV co-infection. Recent studies have shown a promise for the direct detection of pathogen-specific biomarkers such as lipoarabinomannan (LAM) for the diagnosis of TB in HIV-positive individuals. Currently, traditional immunoassay platforms that suffer from poor sensitivity and high non-specific interactions are used for the detection of such biomarkers. In this manuscript, we demonstrate the development of sandwich immunoassays for the direct detection of three TB-specific biomarkers, namely LAM, early secretory antigenic target 6 (ESAT6) and antigen 85 complex (Ag85), using a waveguide-based optical biosensor platform. Combining detection within the evanescent field of a planar optical waveguide with functional surfaces that reduce non-specific interactions allows for the ultra-sensitive and quantitative detection of biomarkers (an order of magnitude enhanced sensitivity, as compared to plate-based ELISA) in complex patient samples (urine, serum) within a short time. We also demonstrate the detection of LAM in urine from a small sample of subjects being treated for TB using this approach with excellent sensitivity and 100% corroboration with disease status. These results suggest that pathogen-specific biomarkers can be applied for the rapid and effective diagnosis of disease. It is likely that detection of a combination of biomarkers offers greater reliability of diagnosis, rather than detection of any single pathogen biomarker.
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1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Cho, Sang Nae(조상래)
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