HIGH-THROUGHPUT SYSTEM DEMONSTRATION FOR FOOT AND MOUTH DISEASE VIRUS DETECTIONIn late July 2006, the CSU Veterinary Diagnostic Lab (CSU VDL) participated in a collaborative effort along with Lawrence Livermore National Laboratory (LLNL), the Department of Homeland Security (DHS), the US Department of Agriculture (USDA) and Animal and Plant Health Inspection Service (APHIS) to demonstrate a high-throughput system for the detection of foot and mouth disease (FMD) virus. Although the last outbreak of foot and mouth disease virus in the United States occurred in 1929, it is still enzootic in many parts of the world.1 Outbreaks create severe economic impacts by halting trade in the cattle industry, creating animal welfare problems, and hindering tourism.1 There are currently no active surveillance systems for FMD2 in the United States and suspect cases currently are tested only at Plum Island. In Colorado, outbreaks of vesicular stomatitis, when affecting cattle, are indistinguishable from FMD. Pre-clinical detection by a method with quick turnaround time, reduced cost, increased efficiency and the ability to differentiate between foot and mouth disease and other clinically similar diseases is key. LLNL, DHS, USDA and APHIS, working with the National Animal Health Laboratory Network (NAHLN) - of which the CSU VDL is a core lab -- developed a multiplexed FMD virus rule-out assay that has the ability to simultaneously detect the presence of viral DNA or RNA for seven different viruses, including those that closely mimic foot and mouth disease, such as bovine herpesvirus, bovine popular stomatitis virus, bovine viral diarrhea virus, bluetongue virus, swine vesicular disease, and vesicular exanthema of swine. This assay also distinguishes between multiple serotypes of each of the viruses listed above including 2 FMDV serotypes (FMDV-1 and FMDV-2). The assay, based on luminex bead technology, hybridizes PCR product extracted from samples to a bead suspension that is labeled with a fluorescent probe. Fluorescence produced by these beads is measured and, based on the individual sample's ratio of red to near-infrared, identifies the presence or absence of the target nucleic acid sequences from the viruses listed above. In September 2005, the CSU VDL was selected as one of 14 labs to participate in an interlaboratory comparison using a manual system and the same technology. Equipment was purchased for each participating lab, and shipped to their facilities. After equipment installation, on-site training by a LLNL staff member took place at each lab. This was followed by an interlaboratory comparison during which each of the participating labs processed and reported results from 200 samples over the course of several days. Following completion of the interlaboratory comparison, LLNL asked for labs that would be interested in further participating in a surge-capacity demonstration. The goal of this demonstration, as stated by the USDA/APHIS, was to "demonstrate to state and federal authorities and laboratory personnel an integrated high-throughput laboratory system capable of processing the surge in sample submissions that would result from a foot and mouth disease outbreak." The CSU VDL and UC Davis CAHFS lab were the two labs chosen to participate in the surge-capacity demonstration. High-throughput automated robotics equipment was installed by LLNL staff in the VDL, and over the course of 8 days, LLNL staff demonstrated the equipment, trained CSU VDL staff on the equipment, and conducted an overview and review of the pilot execution and data collected. On Tuesday, July 25, LLNL staff demonstrated the full capacity of the automated robotics system, processing-from sample receipt by the laboratory through data analysis and results reporting- 1000 samples during a 10-hour day, with two technicians. The following day the CSU VDL staff who had received training on the equipment the week before succeeded in processing 800 samples in a little over 10 hours, demonstrating that in the event of a foreign animal disease outbreak or similar situation, even with minimal training, the equipment and assay could be successfully drop-shipped to approved laboratories, installed, and in use within several days. Not only is such surge capacity ability beneficial during a potential foreign animal disease outbreak, but following an outbreak continued surveillance to establish and prove freedom from the disease may continue for months or even years at high levels. On Thursday, July 27, previously invited guests had the opportunity to tour the laboratory, observe the equipment in operation, and attend scientific presentations related to the demonstration. Interest in this demonstration was overwhelming, with more than 80 participants representing CSU, the Veterinary Medical Center, LLNL, DHS, USDA, AHPIS, the State Veterinarian's office, Senator Ken Salazar's office, Congresswoman Marilyn Musgrave's office, Senator Wayne Allard's office, other laboratories throughout the country, the Colorado Veterinary Medical Association, the Colorado Cattlemen's Association, the Colorado Livestock Association, the National Park Service, various scientific supply companies, and many other organizations. A round-table discussion session in the afternoon allowed participants to ask questions and express possible concerns. Also discussed were future directions and applications including expansion to create bovine and porcine-specific assays and validation of multiple matrices, or sample types, to more closely mimic submission of samples to diagnostic labs. Potential integrated use of the robotics equipment for additional, routine laboratory procedures was also considered. On Friday, July 28, CSU VDL Director Dr. Barb Powers, NAHLN Coordinator for the USDA Barb Martin, deputy director of science for the DHS at the Plum Island Animal Disease Center Dr. Tammy Beckham, and LLNL veterinarian Dr. Pam Hullinger addressed the media, again answering questions, addressing concerns, and providing a tour of the equipment in the lab. Participation in this surge capacity demonstration was an invaluable experience for the CSU veterinary diagnostic lab, giving staff
a chance to work with new equipment, evaluate a new assay, and form and strengthen connections with both community members and other
professionals in the field of veterinary diagnostic medicine. Many thanks to everyone who participated. References: |
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