Rapid and cost-effective virus detection methods using molecular sensors and nano-devices

Document
Description

Accurate virus detection is important for diagnosis in a timely manner to facilitate rapid interventions and treatments. RNA viruses affect an extensive amount of the world’s population, particularly in tropical

Accurate virus detection is important for diagnosis in a timely manner to facilitate rapid interventions and treatments. RNA viruses affect an extensive amount of the world’s population, particularly in tropical countries where emerging infectious agents often arise. Current diagnostic methods have three main problems: they are time consuming, typically not field-portable, and expensive. My research goal is to develop rapid, field-portable and cost sensitive diagnostic methods for RNA viruses. Herein, two different approaches to detect RNA viruses were proposed: Conjugated gold nanoparticles for detection of viral particles or virus-specific antibodies by monitoring changes in their optical properties, and Tentacle Probes coupled with qPCR for detection and differentiation of closely-related viral strains. The first approach was divided into two projects: the study and characterization of the gold nanoparticle-antibody system for detection of virus particles using dynamic light scattering (DLS) and UV-Vis spectrophotometry, and development of a detection method for antibodies using static light scattering (SLS) and antigen-conjugated gold nanoparticles. Bovine serum albumin (BSA) conjugated gold nanoparticles could successfully detect BSA-specific antibodies in vitro, and protein E from Dengue Virus serotype 2 conjugated gold nanoparticles could detect Dengue-specific antibodies, both in vitro and in serum samples. This method is more accurate than currently used detection methods such as dot blots. The second approach uses Tentacle Probes, which are modified molecular beacons, to detect with high specificity two different strains of Lymphocytic Choriomeningitis Virus (LCMV), Armstrong and Clone-13, which differ in only one nucleotide at the target sequence. We successfully designed and use Tentacle Probes for detection of both strains of LCMV, in vitro and in serum from infected mice. Moreover, detection of as little as 10% of Clone-13 strain was possible when diluted in 90% Armstrong strain. This approach enables the detection of different strains of virus even within a mixed quasispecies and may be important for improving intervention strategies for reducing disease. The detection methods provide rapid detection of viruses, including viral strains within mixed populations, and should enhance our ability in providing early responses to emerging infectious diseases due to RNA viruses including Zika or Dengue virus.