Detecting Ebola Virus Infections

DESCRIPTION provided by applicant Ebolavirus attacks are devastating and unpredictable To control an ebolavirus outbreak it is essential to identify isolate and treat infected individuas rapidly However the available diagnostics ELISA and RT PCR require laboratory sophistication that is not possible at point of care in many African and other isolated locations Thus a portable device that needs no laboratory support is essential to detect early markers of ebolavirus infection We propose to develop a point of care detection device targeting the conserved and highly abundant soluble ebolavirus glycoprotein sGP a marker of viral infection that appears in the blood stream early during infection The detection will be enabled by aptamers attached to microcantilevers Aptamers are nucleic acids with many of the properties of antibodies but with the advantage that they are selected and synthesized in vitro and can be reliably reproduced They can be adapted to many assay formats including electrochemical optical colorimetric chemiluminescent andamp fluorescent and mechanical Aptamers are very stable when dehydrated and ideal for sensors that must be stored for long time periods before their use for occasional viral outbreaks As recognition elements for biosensors aptamers have many advantages over antibodies including small size capability for repetitive use long shelf life at room temperature while dehydrated amenability to chemical synthesis and chemical modifications and adaptability to broad assay formats Thus aptamer based biosensors aptasensors are more suitable than antibodies for point of care biosensor development The goal of this proposal is to develop an aptasensor for early diagnosis of ebolavirus infection at point of care either at bedside or at mobile clinic that will lead to effective treatment and containment of the virus outbreak We have three specific aims Use a combination of SELEX next generation sequencing and bioinformatics to identify DNA aptamers that recognize sGPs from all African ebolavirus species with nanomolar affinity Build a prototype array of aptamer functionalized microcantilever device for sGP detection to provide proof of concept for future field application Assess the efficacy and sensitivity of the detection device for recombinant sGP and sGP in serum from infected macaques The innovation of this proposal lies in the integration of aptamer and microcantilever detection technologies which will allow the development of a point of care biosensor for early detection of ebolavirus infection based on the appearance of viral sGP in the blood stream The longer term goal is to expand the capability of the microcantilever aptasensor which can be multiplexed to include the detection of other hemorrhagic viruses that present similar symptoms to ebolavirus in the early stages PUBLIC HEALTH RELEVANCE Ebolavirus outbreaks are devastating unpredictable increasing in frequency result in many lives lost and have the potential for use in terrorism or bio warfare Adequate point of care diagnostics are not yet available particularly for regions of the world lacking sophisticated laboratory equipment We propose to develop an aptamer microcantilever device aptasensor with superior storage qualities and immediate readouts for identifying ebolavirus infected individuals for their immediate treatment and isolation from other susceptible individuals