Development of a Rapid Molecular Diagnostic Platform with Gold Nanoparticle-Based Signal Amplification System for Spinal Infection

Background:The most common bacterial disease throughout the world is tuberculosis, which is caused by Mycobacterium tuberculosis, and the most common pathogen in spinal infection is Staphylococcus aureus. Delayed medical treatment 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, the need for a rapid, simple and accurate diagnostic method to achieve the goal of "early detection and early treatment" has been greater.Signal amplification of biosensors for analytical purposes has attracted much attention in recent years, especially with nanomaterials such as gold nanoparticles. Since the biological samples are limited and difficult to obtain, also in biological samples, matrix compositions are complex and interfering substances are multiple; therefore, the design of biosensors and signal amplification become very crucial for biosensors.Purpose:The purpose of this study is to establish the colorimetric signal amplification methods using gold nanoparticles and two different systems (perAuxidase-TMB-H2O2 colorimetric system and AuNPs-luminol-H2O2 chemiluminescence system), then integrate these two methods on a microfluidic paper-based analytical device. We expect to develop a quick, specific, effective and sensitive biosensing system for the diagnosis of M. tuberculosis and S. aureus in spinal infection, which 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, and the debrided tissue or fluid specimens from patients received spinal surgery for infectious spondylodiscitis (experimental group) and for herniated intervertebral disc (control group). 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 of two different systems: perAuxidase-TMB-H2O2 colorimetric system and AuNPs-luminol-H2O2 chemiluminescence system to detect M. tuberculosis and S. aureus, respectively. In the third year, we will use collected samples to test the microfluidic paper-based analytical device with integrated signal amplification systems, and compare with laboratory culture and TB-PCR results.

Project ID:PC10708-0955
External Project ID:MOST107-2314-B182-015