Viral RNA testing and automation on the bead-based CBNE detection microsystem.

Galambos, Paul C; Bourdon, Christopher Jay; Farrell, Cara M; Rossito, Paul; McClain, Jaime L; Derzon, Mark Steven; Cullor, James Sterling; Rahimian, Kamayar

doi:10.2172/947327

Title: Viral RNA testing and automation on the bead-based CBNE detection microsystem.

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Abstract

We developed prototype chemistry for nucleic acid hybridization on our bead-based diagnostics platform and we established an automatable bead handling protocol capable of 50 part-per-billion (ppb) sensitivity. We are working towards a platform capable of parallel, rapid (10 minute), raw sample testing for orthogonal (in this case nucleic acid and immunoassays) identification of biological (and other) threats in a single sensor microsystem. In this LDRD we developed the nucleic acid chemistry required for nucleic acid hybridization. Our goal is to place a non-cell associated RNA virus (Bovine Viral Diarrhea, BVD) on the beads for raw sample testing. This key pre-requisite to showing orthogonality (nucleic acid measurements can be performed in parallel with immunoassay measurements). Orthogonal detection dramatically reduces false positives. We chose BVD because our collaborators (UC-Davis) can supply samples from persistently infected animals; and because proof-of-concept field testing can be performed with modification of the current technology platform at the UC Davis research station. Since BVD is a cattle-prone disease this research dovetails with earlier immunoassay work on Botulinum toxin simulant testing in raw milk samples. Demonstration of BVD RNA detection expands the repertoire of biological macromolecules that can be adapted to our bead-based detection. The resources of thismore »« less

Authors:

Galambos, Paul C; Bourdon, Christopher Jay; Farrell, Cara M; Rossito, Paul ^[1]; McClain, Jaime L; Derzon, Mark Steven; Cullor, James Sterling ^[1]; Rahimian, Kamayar

University of California at Davis

Publication Date:: Mon Sep 01 00:00:00 EDT 2008

Research Org.:: Sandia National Laboratories (SNL), Albuquerque, NM, and Livermore, CA (United States)

Sponsoring Org.:: USDOE

OSTI Identifier:: 947327

Report Number(s):: SAND2008-7332
TRN: US200906%%57

DOE Contract Number:: AC04-94AL85000

Resource Type:: Technical Report

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; ANIMALS; AUTOMATION; AVAILABILITY; CATTLE; CHEMISTRY; DETECTION; DIARRHEA; DISEASES; IMMUNOASSAY; MODIFICATIONS; NUCLEIC ACID HYBRIDIZATION; NUCLEIC ACIDS; RNA; SENSITIVITY; TESTING; TOXINS; Biochemistry.; Nucleic acids-Analysis; Chemical sensors.; Microelectromechanical devices-Technological innovations

Citation Formats


                    Galambos, Paul C, Bourdon, Christopher Jay, Farrell, Cara M, Rossito, Paul, McClain, Jaime L, Derzon, Mark Steven, Cullor, James Sterling, and Rahimian, Kamayar. Viral RNA testing and automation on the bead-based CBNE detection microsystem..  United States: N. p., 2008. 
        Web.  doi:10.2172/947327.

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                    Galambos, Paul C, Bourdon, Christopher Jay, Farrell, Cara M, Rossito, Paul, McClain, Jaime L, Derzon, Mark Steven, Cullor, James Sterling, & Rahimian, Kamayar. Viral RNA testing and automation on the bead-based CBNE detection microsystem..  United States.  https://doi.org/10.2172/947327

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                    Galambos, Paul C, Bourdon, Christopher Jay, Farrell, Cara M, Rossito, Paul, McClain, Jaime L, Derzon, Mark Steven, Cullor, James Sterling, and Rahimian, Kamayar. 2008.  
        "Viral RNA testing and automation on the bead-based CBNE detection microsystem.".  United States.  https://doi.org/10.2172/947327.  https://www.osti.gov/servlets/purl/947327.

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@article{osti_947327,

  title        = {Viral RNA testing and automation on the bead-based CBNE detection microsystem.},

  author       = {Galambos, Paul C and Bourdon, Christopher Jay and Farrell, Cara M and Rossito, Paul and McClain, Jaime L and Derzon, Mark Steven and Cullor, James Sterling and Rahimian, Kamayar},

  abstractNote = {We developed prototype chemistry for nucleic acid hybridization on our bead-based diagnostics platform and we established an automatable bead handling protocol capable of 50 part-per-billion (ppb) sensitivity. We are working towards a platform capable of parallel, rapid (10 minute), raw sample testing for orthogonal (in this case nucleic acid and immunoassays) identification of biological (and other) threats in a single sensor microsystem. In this LDRD we developed the nucleic acid chemistry required for nucleic acid hybridization. Our goal is to place a non-cell associated RNA virus (Bovine Viral Diarrhea, BVD) on the beads for raw sample testing. This key pre-requisite to showing orthogonality (nucleic acid measurements can be performed in parallel with immunoassay measurements). Orthogonal detection dramatically reduces false positives. We chose BVD because our collaborators (UC-Davis) can supply samples from persistently infected animals; and because proof-of-concept field testing can be performed with modification of the current technology platform at the UC Davis research station. Since BVD is a cattle-prone disease this research dovetails with earlier immunoassay work on Botulinum toxin simulant testing in raw milk samples. Demonstration of BVD RNA detection expands the repertoire of biological macromolecules that can be adapted to our bead-based detection. The resources of this late start LDRD were adequate to partially demonstrate the conjugation of the beads to the nucleic acids. It was never expected to be adequate for a full live virus test but to motivate that additional investment. In addition, we were able to reduce the LOD (Limit of Detection) for the botulinum toxin stimulant to 50 ppb from the earlier LOD of 1 ppm. A low LOD combined with orthogonal detection provides both low false negatives and low false positives. The logical follow-on steps to this LDRD research are to perform live virus identification as well as concurrent nucleic acid and immunoassay detection.},

  doi          = {10.2172/947327},

  url          = {https://www.osti.gov/biblio/947327},
  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {Mon Sep 01 00:00:00 EDT 2008},

  month        = {Mon Sep 01 00:00:00 EDT 2008}

}

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