The University of New Mexico

Sandia Corporation

National Technology & Engineering Solutions of Sandia, LLC

Sandia National Laboratories

Brian Hjelle

David R Wheeler

Darren W Branch

Thayne L Edwards

Richard Smith Larson

David C Brown

Susan M Brozik

Marco Bisoffi

Steve Jenison

Pam R Hall

Viruses and other bioagents are of high medical and biodefense concern and their detection at concentrations well below the threshold necessary to cause health hazards continues to be a challenge with respect to sensitivity, specificity, and selectivity. Ideally, assays for accurate and real time detection of viral agents and other bioagents would not necessitate any pre-processing of the analyte, which would make them applicable for example to bodily fluids (blood, sputum) and man-made as well as naturally occurring bodies of water (pools, rivers). We describe herein a robust biosensor that combines the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325 MHz with the specificity provided by antibodies and other ligands for the detection of viral agents. In preferred embodiments, a lithium tantalate based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against Coxsackie virus B4 or the negative-stranded category A bioagent Sin Nombre virus (SNV), a member of the genus Hantavirus, family Bunyaviridae, negative-stranded RNA viruses. Rapid detection (within seconds) of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, although the sensor was approximately 50&#xd7;10-fold more sensitive for the detection of SNV. For both pathogens, the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1. The biosensor was able to detect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS). Further, in a proof-of-principle real world application, the SAW biosensor was capable of selectively detecting SNV agents in complex solutions, such as naturally occurring bodies of water (river, sewage effluent) without analyte pre-processing.

Detection of bioagents using a shear horizontal surface acoustic wave biosensor

A biosensor combining the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325 MHz with the specificity provided by antibodies and other ligands for the detection of viral agents. In a preferred embodiment, a lithium tantalate based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against Coxsackie virus B4 or the negative-stranded category A bioagent Sin Nombre virus (SNV). Rapid detection of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, and the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1 The biosensor was able to delect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS).

The invention provides HARDS virus rDNA for expression in molecular clones. The expressed products are useful in immunodiagnostics, prophylactics, and therapeutics for the HARDS virus and related hantaviruses. Of particular interest are a type-specific epitope of the HARDS virus G1 protein, and dominant epitopes of the HARDS virus N protein cross-reactive with antibodies to the HARDS virus and the related hantavirus PHV, both expressed by cDNA clones according to the invention.

Molecular clones producing recombinant DNA antigens of the hantavirus-associated respiratory distress (HARDS)

Hantavirus-associated respiratory distress virus antigens

Growing community of inventors

Placitas, NM, United States of America

Brian Hjelle