Recipient Organization
UNIVERSITY OF ILLINOIS
2001 S. Lincoln Ave.
URBANA,IL 61801
Performing Department
(N/A)
Non Technical Summary
In this project, we will develop and test a diagnositc testing technology that can be performed on a blood sample from an animal, providing results in <30 minutes to determine if the animal is infected with African Swine Fever Virus (ASFV). The test will be divided into two parts, in which one test will detect the presence of anti-ASFV antibody (to determine if the animal was previously infected) and the secnod test will detect the presence of DNA from the ASFV virus. The project will use a new biosensing technology platform in whichmolecules are tagged with gold nanoparticles and the nanoparticles are counted with a small, inexpensive instrument. The method can be extended for determining infection status for other pathogens of concern.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Goals / Objectives
The overall goal of this project is to develop portable, rapid, sensitive, and inexpensive point-of-use biosensor technology and assays for on-farm detection of ASFV infection in blood samples in aid of field surveillance and outbreak diagnosis.Objective 1: To develop nanosensor-based assays to detect ASFV DNA and anti-ASFV antibody in laboratory settings.Objective 2: To adapt laboratory assays into a field-deployable platform.Objective 3. To validate the nanosensor-based assays using samples from ASFV-infected animals.
Project Methods
1. The PRAM detection instrument utilizes optical design principles for selection and alignment of components (LED, microscope objective, image sensor, optical filters), mechanical design using computer aided design tools (for construction of the enclosure, sample holding stage, 3D printed component holding fixtures), electronic design (for selection and interconnection of microcontroller, BlueTooth circuit board, power management circuit, and rechargeable battery), and softwre.2. The microfluidic cartridge utilizes fluid dynamic simulation software (COMSOL Multiphysics) and mechanical CAD software (AutoCAD) for design of dimensions. 3D printing (FormLabs) is used for fabricating designed components at prototyple scale. Completed cartridges are assembled from their components (3D printed parts, o-rings, adhesive tapes), and some parts are cut to precise dimensions by laser cutting.3. Assays are performed using reagents that are obtained from standard commercially avilable sources (gold nanoparticles from Cytodiagnostics) combined with special-purpose reagents (monoclonal antibodies) developed in our own facility and nucleic acid probe molecules that are synthesized with our specifiedbase sequences by a commercial vendor (IDT).4. Assays will be performed at least in triplicate to determine coefficient of variation values at all tested concentrations, and Receiver Operating Characteristic (ROC) analysis will be used to determine sensitivity and selectivity of the test, via analysis of false positive and false negative rates when using the test upon blood from infected or healthy animals.