Development of a Rapid Molecular Sensing Platform Using Fluorescent Nanomaterials for Clinical Diagnosis of Bacterial Infection

Background: Tuberculosis, which is caused by Mycobacterium tuberculosis, is one f the most common bacterial disease throughout the world, and Staphylococcus aureus is the most common pathogen in spinal infection. Delayed medical treatment for spinal infection may lead to paralysis of lower extremity and sepsis. However, the traditional detection methods rely on high-end instruments or prolonged bacterial culture with the chance of resulting in false negatives and false positives. Therefore, there’s a need for a rapid, simple and accurate diagnostic method to achieve the goal of "early detection and early treatment". Signal amplification of biosensors for analytical purposes has attracted much attention in recent years, especially with noble metal nanomaterials such as gold, silver, copper, and platinum. Since the biological samples are limited and difficult to obtain, and also have a highly complex matrix compositions with multiple interfering substances; therefore, the design of biosensors and signal amplification has become very crucial for biosensors. Purpose: The purpose of this study is to develop a rapid molecular sensing platform using the rolling circle amplification and loop-mediated isothermal amplification combined with fluorescent nanomaterials to detect bacterial infection such as Mycobacterium tuberculosis and Staphylococcus aureus. We expect this quick, specific, effective and sensitive biosensing system will meet the ASSURED (affordable, sensitive, specific, user-friendly, rapid and robust, equipment-free and deliverable to end-users) criteria, outlined by World Health Organization. Materials and Methods: Throughout the entire 3-year study period, we will collect the delinked residual sample with TB-PCR results. Also, we will use the collected synovial fluid samples of patients received revision surgery for PJI (experimental group) and for aseptic loosening or implant wear debris (control group) after hip or knee arthroplasties from previous study. The medical charts including patient demographic data and laboratory reports of enrolled patients are also reviewed, and later compared with experimental results. In the first 2 years of study, we will test the established signal amplification assays using two different systems (rolling circle amplification and loop-mediated isothermal amplification) and both combined with fluorescent nanomaterials to detect M. tuberculosis and S. aureus, respectively. In the third year, we will use collected samples to test the molecular sensing platform with integrated signal amplification systems, and compare with laboratory culture and TB-PCR results.

Project ID:PC10901-1630
External Project ID:MOST108-2314-B182-025-MY3