RAPID: Improved Detection of Ebola through Nanomanufacturing of Bio-Inspired Diagnostics

Case Western Reserve University, Cleveland OH

Investigators

Abstract

This Rapid Response Research (RAPID) grant will pioneer a novel diagnostic tool that will enable the detection of the deadly Ebola virus with high accuracy through reduction of false negative results. The test-bed developed through bioengineering design and nanomanufacturing will be implemented in high-tech and low-tech detection assays. The latter will enable detection on-site and in-field, testing human specimens, livestock and agricultural samples without the need for specialized equipment. The ongoing Ebola epidemic could potentially be the largest in history, not only affecting multiple countries in West Africa, but also U.S. and other healthcare personnel working on-site in disease management and containment. While therapeutics and vaccines are undergoing development and testing, protection or treatment of this life-threatening virus is not yet available. Therefore, detection of Ebola virus is the first step, and currently the only option, to prevent the spread of the disease. Results from this research will benefit the economy and society on a global scale. An important further benefit is the training and education of scholars in an interdisciplinary environment fostering team sciences, bridging bioengineering, nanomanufacturing, virology, and molecular biology. K-12 and public engagement will be promoted through the established 'The Nanoman' outreach program. Reverse transcription polymerase chain reaction is a molecular biological technique used to detect Ebola virus in diverse specimens. While this method has the potential to be highly sensitive, the assays lack internal controls to negate false negative results due to processing errors. Contemporary controls are synthetic RNA transcripts. These 'naked' RNA transcripts are unstable and prone to degradation, therefore these molecules cannot be reliably used to spike samples under investigation; the lack of verification of negative results presents a considerable health and environmental risk. This research sets out to overcome this technological hurdle through bioengineering design and nanomanufacturing of an intrinsic positive control, in which the synthetic RNA transcripts are packaged into a plant virus capsid. Such a probe mimics more closely the conditions encountered by the RNA template of the Ebola virus within clinical or environmental samples. The diagnostic probe can be added to each specimen prior to processing and will reveal whether various sample-processing steps have resulted in degradation of the Ebola virus target and resulted in a false negative read-out. In response to the urgency of the Ebola epidemic, the diagnostic probe will be implemented in high-tech and low-tech assays to enable accurate detection in hospitals and in-field and on-site.