Developing a Surface Antigen Detection Device for Tuberculosis Using Rv3807c/Rv3887c Transmembrane Proteins with Paper-Based Colorimetric Gold Nanoparticles

Background & Purpose: Tuberculosis (TB) is a common and often lethal infectious disease caused by various strains of mycobacteria, usually Mycobacterium tuberculosis in humans. It has always been a global public health problem. If not treated, spinal tuberculosis may cause to spinal deformity, or even paraplegia, and pulmonary insufficiency. Diagnosing TB is difficult, especially hard to distinguish between TB infection, other bacterial infections and spinal tumor from X-ray, CT and MRI assessments. Gold nanoparticles (AuNPs) have recently attracted widespread attention due to their unique electronic and optical properties; they have been widely used for diagnostics and increasing used in the field of therapeutics. There are several advantages using nanoparticles in aggregation-based immunoassay. In order to develop a rapid diagnostic tool for TB, we plan to directly detect the surface antigen of Mycobacterium tuberculosis rather than detecting the DNA. The purpose of this study is to investigate the efficiency of using Rv3807c and Rv3887c transmembrane proteins to produce antibody to detect the surface antigen of TB. Then, integrate them into gold nanoparticles, so TB can be easily and quickly detected using aggregation-based immunoassay through color change, which allows surgeons to make rapid and accurate diagnosis and treatment. Materials and Methods: Patients with spinal tuberculosis often required surgical intervention to prevent neurologic deficits and correct spinal deformities in advanced cases. Surgically debrided tissue samples are collected to diagnose TB by detecting the surface antigen of Mycobacterium tuberculosis. This study will only use debrided tissues that were soaked in formaldehyde to inactivate bacteria for infection control, so it will not affect the treatment of patients, and it also will not involve any bacterial cultures. The first year of the study: We plan to test the antibody derived from Rv3807c and Rv3887c transmembrane proteins on histological slides sectioned from formalin fixed paraffin wax embedded disc tissue samples. The tissue samples were surgically harvested from patients with TB and stored at the Tissue Bank. Immunohistochemistry is performed to evaluate the specificity of antibody for TB, and immunostaining is examined under light microscope. In addition, western blotting and ELISA are also performed. The second year of the study: We plan to develop a surface antigen detection device for tuberculosis with Rv3807c/Rv3887c transmembrane proteins and paper-based colorimetric gold nanoparticles. In order to find the minimal concentration of DNA and optimal size of gold nanoparticles, we are going to perform aggregation-based immunoassay with gold nanoparticles to test the surface antigen Rv3807c/Rv3887c on harvested tissues, and record the color change of solutions. The efficiency of using gold nanoparticles as a diagnostic tool for TB is investigated by comparing its sensitivity and specificity results with other tests such as PCR, smear, culture and pathology. The possibility of using gold nanoparticles on surgically debrided tissues directly is also examined.

Project ID:PC10308-1714
External Project ID:MOST103-2314-B182-034