, 925-422-5545, smith350@llnl.gov Contributors:Linda Danganan, 925-422-2553, danganan1@llnl.gov Lance Tammero, PhD., 925-423-9545, tammero2@llnl.gov Beth Vitalis, PhD., 925-422-0149, vitalis1@llnl.gov Chemical and Biological Countermeasures Division Nonproliferation, Homeland and International Security (NHI)Lawrence Livermore National Laboratory (LLNL) Livermore, CA Principle Investigators: Ray Lenhoff, Ph.D., PhD., 925.422.5665, pnaraghi@llnl.gov Benjamin Hindson, PhD., hindson1@llnl.gov This work was performed under the auspices of the U.S. . This effort has the ability to improve our nation's capability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect food and agricultural resources with a diagnostic test which could enhance the nation's capabilities for early detection of a foreign animal disease.In FY2005 A timeline for this development is presented in Table 1. The development of the Version 1.0 panel for FMDV rule-out and the most current efforts aimed to designed species specific panels has spanned over 2 ½ years with multiple collaborative partnerships.This document provides a summary of the development, testing and performance data at OIE Stage 1 Feasibility into Stage 2 Assay Development and Standardization 1 (see Table 2), gathered as of June 30 th , 2007 for the porcine and bovine MUX assay panels. We present an overview of the identification and selection of candidate genetic signatures, the assay development process, and preliminary performance data for each of the individual signatures as characterized in the multiplexed format for the porcine and bovine panels. The Stage 1 Feasibility data of the multiplexed panels is presented in this report also includes relevant data acquired from the Version 1.0 panel as supporting information where appropriate. In contrast to last years' effort, the development of the bovine and porcine panels is pending additional work to complete analytical characterization of FMDV, VESV, SVD, RPV and MCF. The signature screening process and final panel composition impacts this effort. The unique challenge presented this year was having strict predecessor limitations in completing characterization, where efforts at LLNL must precede efforts at PIADC, such challenges were alleviated in the 2006 reporting by having characterization data from the interlaboratory comparison and at Plum Island under AgDDAP project. We will present an addendum at a later date with additional data on the characterization of the porcine and bovine multiplex assays when that data is available. As a summary report, this document does not provide the details of 90% complete: during development, each MUX panel was tested against viral nucleic acid extracts (total nucleic acids, non-titered) from each organism. Preliminary results assessed by low-resolution titration curves, for each step-wise addition of assay to the multiplexed panel. 1.2.1.2.Calibration test of reference standards 90% complete: refere...
Primary Author:Sally Smith, 925-422-5545, smith350@llnl.gov Co-authors:Linda Danganan, 925-422-2553, danganan1@llnl.gov Lance Tammero, PhD., 925-423-9545, tammero2@llnl.gov Chemical and Biological National Security Program Lawrence Livermore National Laboratory (LLNL) Livermore, CA Principle Investigators: Ray Lenhoff, Ph.D., PhD., 925.422.5665, pnaraghi@llnl.gov Benjamin Hindson, PhD., hindson1@llnl.gov This work was performed under the auspices of the U.S. Department of Energy by the University of California, Lawrence Livermore National Laboratory under Contract No. W-7405-Eng-48, with funding through the Department of Homeland Security (DHS). Distribution authorized to US government agencies and their contractors for administrative and operational use.This document was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor the University of California nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States Government or the University of California. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States Government or the University of California, and shall not be used for advertising or product endorsement purposes. Bioassays and Signatures Program Page 1 of 489Ag Assay Development: FMDV Rule-out panel Ag Assay Development: FMDV Rule-out panel Report Executive summaryLawrence Livermore National Laboratory (LLNL), in collaboration with the Department of Homeland Security (DHS) and the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Services (APHIS) has developed advanced rapid diagnostics that may be used within the National Animal Health Laboratory Network (NAHLN), the National Veterinary Services Laboratory (Ames, Iowa) and the Plum Island Animal Disease Center (PIADC). This effort has the potential to improve our nation's ability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect animal populations of high economic importance in the United States. This document provides details of signature generation, evaluation, and testing, as well as the specific methods and materials used. A condensed summary of the development, testing and performance of the multiplexed assay panel was presented in a 126 page separate document, entitled "Development and Characterization of A Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out". This supp...
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