AAVLD Annual Conference 2016
Visit us at booths 219, 221 and 223, October 13th-19th
We look forward to seeing you at the American Association of Veterinary Laboratory Diagnosticians (AAVLD) Conference 2016, and sharing with you some of our leading-edge solutions for your lab.
Don’t miss our informative presentations, poster, and workshop (see details below). Also, don’t forget to visit us at booth #219–223: stop by and play the interactive trivia game to receive a free gift. You can also register for a chance to win a Bose™ SoundLink™ Color Bluetooth Speaker.*
Speaker: Angela Burrell, Staff Scientist, Thermo Fisher Scientific
Date & Time: Sunday October, 16 at 10:15 am ET
Location: Imperial F
Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent for Johne’s disease in cattle and causes severe economic losses in the cattle industry due to reduced productivity, reproductive losses, and the eventual death or culling of the infected animal.
The new Applied Biosystems™ VetMAX™-Gold MAP Detection Kit is a real-time PCR assay for the rapid in vitro detection of MAP DNA purified from bovine feces. The assay targets a unique sequence element in the MAP genome to enable highly sensitive and specific results. The purpose of this study is to determine the performance characteristics of the VetMAX-Gold MAP Detection Kit in detecting MAP DNA from nucleic acid extracted from individual and pooled bovine fecal samples.
The kit was evaluated with 126 individual MAP-positive and 134 individual MAP-negative bovine fecal samples. The MAP status of each sample was confirmed with culture prior to the start of the study. MAP samples were sourced from diverse geographic regions and represented a range of MAP infectivity, including heavy, moderate, and light shedders.
The feasibility of pooling up to 5 bovine fecal samples into a single nucleic acid extraction and detection test was evaluated by testing 51 MAP-positive pools and 24 MAP-negative pools. All pools consisted of 5 individual fecal samples. 49 positive pools were created by combining 1 MAP-positive sample with 4 MAP-negative samples. 2 positive pools were created by combining 2 MAP-positive samples with 3 MAP-negative samples.
A collaborator laboratory purified nucleic acid from bovine fecal samples using the Applied Biosystems™ MagMAX™ Total Nucleic Acid Isolation Kit. MAP bacterium was physically and chemically lysed by homogenizing the fecal supernatant samples using the FastPrep-24™ homogenizer in the presence of lysis solution. 5,000 copies/reaction of Applied Biosystems™ Xeno™ DNA Control were spiked into the lysis solution of each purification to monitor for inhibition. Samples were processed using the Applied Biosystems™ MagMAX™ Express-96 Deep Well Magnetic Particle Processor. 8 µL of extracted nucleic acid was tested using the VetMAX-Gold MAP Detection Kit on the Applied Biosystems™ 7500 Fast Real-Time PCR System according to the Instructions for Use.
The results of testing show the VetMAX-Gold MAP Detection Kit produced diagnostic sensitivity and specificity values of 96.2% and 96.4%, respectively, when testing individual fecal samples as compared to culture. Pooled sample testing with the VetMAX-Gold MAP Detection Kit resulted in diagnostic sensitivity and specificity values of 96.2% and 100%, respectively, as compared to culture. This study indicates that DNA isolated from diagnostic bovine fecal sample, tested with the VetMAX-Gold MAP Detection Kit, provides an economical and rapid solution for MAP detection from both individual and pooled fecal samples. The results of this study are under review by APHIS Center for Veterinary Biologics in support of a Biological Product License application.
Speaker: Rick Conrad, Sr. Manager, R&D, Thermo Fisher Scientific
Date & Time: Sunday, October 16, 2016 at 11:15 AM ET
Location: Imperial F
qPCR/qRT-PCR are the most sensitive high-throughput technologies for the diagnosis of infections in food animals. A common reason for false negatives is molecular inhibitors in the biological sample. Inhibition is mitigated by chemical characteristics of the reaction master mix. Consequently, animal health (AH) customers are vigilant about the performance of master mixes. To address this need, we conducted a performance study of 10 master mixes in the Thermo Fisher portfolio alongside 10 of our leading competitors.
We extracted five common biological sample types encountered by our customers. Into each extract, we spiked a dilution series of positive control and assay to detect a porcine virus (for qRT-PCR) or a bovine bacterium (for qPCR). We measured master mix performance by detection sensitivity. For both qPCR and qRT-PCR, Applied Biosystems™ Path-ID™ master mixes performed the best overall out of the Thermo Fisher products, and performed equal to or better than external competitors. However, the most interesting results were individual performance by sample type: for example, a master mix that is superior for blood samples is not necessarily one to use for saliva samples.
This study directly benefits our customers by providing guidelines for master mix use for specific sample types, as well as for overall performance.
Speaker: Robert Tebbs, Sr. Manager, R&D, Thermo Fisher Scientific
Date & Time: Saturday October, 15 at 2:00 pm ET
Location: Imperial H
Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine delta coronavirus (PDCoV) cause gastrointestinal diseases in pigs. Clinical signs can include diarrhea, vomiting, dehydration and anorexia. The diseases are often associated with villous atrophy.
The emergence of PEDV and PDCoV in the US in May 2013 and January 2014, respectively, caused the deaths of approximately 7 million pigs in the first year, mainly among piglets (1). We designed a real-time, reverse transcriptase PCR (RT-PCR) assay to detect all three coronaviruses in a single reaction. The multiplex RT-PCR assay uses a different fluorescent dye for each target for pathogen ID, and includes an internal positive control. The assay was designed to detect all PEDV, PDCoV, and TGEV sequences found in GenBank and tested on field samples to validate sensitivity and specificity. The assay showed an analytical sensitivity of ≤10 copies for all three targets when present individually, and ≤80 copies in complex samples when the concentration of the other two targets was >106 copies.
The sensitivity and specificity of the multiplex coronavirus assay was determined on field samples consisting of oral fluids, feces, and environmental samples. All field samples were purchased from Iowa State University, the University of Minnesota, or Rural Technologies. Over 400 field samples were tested. The workflow for testing field samples includes RNA isolation, RT-PCR, and data analysis. Total RNA was isolated from field samples using the Applied Biosystems™ MagMAX™ Pathogen RNA/DNA Kit. The RT-PCR reaction was prepared by combining the Applied Biosystems™ TaqMan® Fast Virus 1-Step Master Mix, the coronavirus primer/probe mix, water, and 8 µL of the isolated RNA sample (20 µL total reaction volume). The samples were run on the 7500 Fast Real-Time PCR System using standard run mode with the following cycling conditions: 48°C for 10 min, 95°C for 10 min, followed by 40 reps at 95°C for 15 seconds and 60°C for 45 seconds. The data were analyzed by setting the threshold as a percentage of thepositive control reactions (control-based threshold). Results with a Ct value ≤37 are classified positive; results between 37 and 40 are suspect and require repeat testing; and undetected results (≥40 Ct) are classified negative. Results from testing field samples showed a sensitivity of ≥97% and a specificity of 100% for all three targets.
In conclusion, a multiplex RT-PCR assay was designed for detecting three enteric coronaviruses. Here we show a workflow that is simple, sensitive, and accurate for detecting PEDV, PDCoV, and TGEV in diverse biological and environmental samples.
Speakers: Michelle Swimley, Rohan Shah, and Rick Conrad
Misdiagnosis of one sick animal can endanger the health of an entire herd, affecting the producer’s livelihood and potentially damaging a lab’s reputation. When using real-time PCR for molecular diagnostic testing, false negatives rarely result from a lack of sensitivity. Rather, the biological material used to test these animals can contain a variety of compounds that inhibit the actual PCR chemistry. A simple way to test for PCR inhibition is to include an internal positive control. Applied Biosystems™ VetMAX™ Xeno™ Internal Positive Control (IPC) products for both DNA and RNA testing can be easily integrated into any real-time PCR workflow, greatly reducing the likelihood of costly false negatives and thus providing confidence in the accuracy of our customer’s qPCR test results.
Although Xeno chemistry has been incorporated into many of our existing animal health diagnostic products, these products, which include Xeno IPC templates and Xeno IPC assays, are now conveniently available as standalone products that are directly orderable from our catalog. The Xeno IPC assay is a highly specific primer/probe mix that detects the Xeno IPC and is available in VIC and LIZ dye channel formats for flexibility of multiplexing with a variety of diagnostic assays. The Xeno IPC template, available as either DNA or RNA, was designed as a unique synthetic sequence and was evaluated against a comprehensive nucleotide database to show no off-target alignments to sequences relevant to animal health. The Xeno IPC template serves as both an internal positive control for the recovery of nucleic acid during the isolation process, and as a positive control for qPCR. It is a very effective control for qPCR inhibition, which enables the customer to be easily alerted to problematic test results, helping to reduce the likelihood of false-negatives.
Xeno IPC performance was tested and shown to be compatible with a wide variety of veterinary sample matrices and qPCR master mixes. Nucleic acid isolation was performed using the MagMAX Total Nucleic Acid Isolation Kit with an automated MagMAX Express-96 instrument. Real-time PCR was performed on a 7500 Fast Real-Time PCR System and on an Applied Biosystems™ QuantStudio™ 5 Real-Time PCR System.
Recommended by the American Association of Veterinary Laboratory Diagnosticians (AAVLD) and used in our USDA-licensed kits, Xeno IPC is a trusted product intended for use in a growing animal health diagnostic market seeking improved quality control.
Speaker: Derek Grillo, Thermo Fisher Scientific Product Manager, Animal Health
Date & Time: Saturday, October 15, 6:30–7:00 p.m. ET
Location: Imperial B
Accelerating today’s monitoring of animal health is what you do best. Advanced diagnostic testing is how you get there. Dependable chemistry, flexible platforms, government-licensed testing solutions–they’re all critical in setting new farm animal testing standards. And it all must work together, rapidly and accurately, because timely diagnosis of just one sick animal can help ensure the health of an entire herd, protect a producer's livelihood, and safeguard a lab's business. The Applied Biosystems AgriBusiness team is innovating workflows that help labs stay ahead of the evolving animal disease landscape, such as:
- VetMAX Xeno IPC products: Novel solutions for lab-designed assays, designed to offer a verification layer to help ensure that the qPCR test results delivered to customers are accurate and actionable
- QuantStudio 3 & 5 Systems: The next-generation qPCR instrumentation for users who need an affordable, easy-to-use qPCR system without compromising on performance and quality
- Thermo Fisher Cloud: A state-of-the-art, scientific computing & analysis ecosystem, which enables users to access and share data easily and securely
All workshop attendees will receive a free portable USB charger.**
Short Terms & Conditions
* NO PURCHASE NECESSARY TO ENTER OR WIN. This promotion is open only to registered attendees of AAVLD 2016 (excluding vendors) who are 21 years and older. Health care professionals (term defined in the complete terms and conditions) may not participate in this promotion. Eligible participants who play the interactive trivia game (the “Game”) at Sponsor’s booth #219, 221, and 223 must stop by the booth no later than 1:00 p.m. on October 16, 2016 to be included in the Giveaway. Each participant will receive a cow USB device (approx. retail value $6.55 USD) after arrival at the booth. The participant who completes the Game with the fastest time each day (October 15 and 16) will receive a Bose™ SoundLink™ Color Bluetooth Speaker(approx. retail value $116.99). By playing the Game, you warrant that you are not prohibited by employment, contract, or law from accepting a prize from the Sponsor. Offer void where prohibited, licensed, or restricted by federal, state, provincial, or local laws, regulations, or agency/institutional policies. Other restrictions may apply. Complete Terms and Conditions are available at the Sponsor's booth.
** NO PURCHASE NECESSARY TO ENTER OR WIN. This promotion is open only to registered attendees of AAVLD 2016 (excluding vendors) who are 21 years and older. Health care professionals (term defined in the interactive trivia game complete terms and conditions available at Sponsor's booth) may not participate in this promotion. The workshop free charger(approx. retail value $10USD) giveaway will take place on October 15, 2016. Participants must attend the workshop no later than 6:35 p.m. on October 16, 2016 to be included in the Giveaway. By attending the workshop, you warrant that you are not prohibited by employment, contract, or law from accepting a prize from the Sponsor. Offer void where prohibited, licensed, or restricted by federal, state, provincial, or local laws, regulations, or agency/institutional policies. Other restrictions may apply.
NO PURCHASE NECESSARY TO ENTER OR WIN.
HOW TO ENTER: Stop by the Thermo Fisher Scientific booth (booth #219,221,223) and get a free gift. (cow USB) The fastest time each day will receive a Bose® - SoundLink® Color Bluetooth Speaker $116.99.
ELIGIBILITY: Participation is open only to registered attendees (excluding vendors) of the American Association Of Veterinary Laboratory Diagnosticians (AAVLD) who are 21 years or older and who play the interactive trivia game. Healthcare Professionals may not participate in this promotion. The term Healthcare Professionals includes individuals (clinical or non-clinical physicians, physician assistants, nurses, technicians, research coordinators, administrators, and purchasing personnel) and entities (including hospitals, rehabilitation centers, nursing facilities, home health agencies, clinics and group purchasing organizations, managed care organizations, physician group practices, medical directors for health insurance organizations) and their employees and representatives who are involved in the actual provision of health care services.
DURATION: The booth giveaway begins October 15th & 16th, 2016. Participants must stop by the booth at booth 219, 221, and 223 no later than October 16th, 2016 at 1:00 p.m. to be included in the Giveaway.
WINNER: Each participant will receive a cow USB after arrival at the booth. The fastest time each day will receive a Bose® - SoundLink® Color Bluetooth Speaker $116.99. The participant must warrant that she or he is not a Healthcare Professional, nor prohibited by employment, contract, or law from entering the Sweepstakes or accepting a prize from the Sponsor.
PRIZE: Every participant will receive a cow USB (retail value approximately $6.55 USD). No cash or other substitutions are permitted, except by the Sponsor, which reserves the right to substitute the prize with one of equal or greater value. No refunds or credits are allowed. Prize is not transferable. All taxes on the prize and reporting thereof are the sole responsibility of the Winner.
By entering the Giveaway, each participant unconditionally accepts and agrees to comply with and abide by these rules and the decisions of the Sponsor, which shall be final and binding in all respects.
PUBLICITY: Sponsor will not publish the name of the participants.
INDEMNITY; DISCLAIMERS: To the full extent allowed by law, participants and each Winner release, indemnify and hold harmless the Sponsor and its affiliates, directors, officers and employees from all liability for any damages, injury or losses to any person (including death and bodily injury) or property of any kind resulting in whole or in part, directly or indirectly, from acceptance, possession, misuse or use of the prize or participation in any promotion-related activity or participation in this promotion. The Sponsor has not made any warranty, representation, or guarantee, express or implied, in fact or in law, with respect to any prize, including, without limitation, to such prize’s quality or fitness for a particular purpose. IN NO EVENT SHALL THE SPONSOR BE LIABLE, WHETHER IN CONTRACT, TORT, WARRANTY, OR UNDER ANY STATUTE OR ON ANY OTHER BASIS FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES SUSTAINED BY ANY WINNER OR ANY OTHER PERSON OR ENTITY ARISING OUT OF THE SPONSOR'S PERFORMANCE OR FAILURE TO PERFORM ITS OBLIGATIONS RELATING TO THE AWARD OF THE PRIZE OR PERFORMANCE OF SERVICES, THE POSSESSION OR USE OF ANY PRODUCT, OR THE PERFORMANCE BY THE SPONSOR OF ANY SERVICES, WHETHER OR NOT FORESEEABLE AND WHETHER OR NOT THE SPONSOR IS ADVISED OF THE POSSIBILITY OF SUCH DAMAGES, INCLUDING WITHOUT LIMITATION DAMAGES ARISING FROM OR RELATED TO LOSS OF USE, LOSS OF DATA, DOWNTIME, OR FOR LOSS OF REVENUE, PROFITS, GOODWILL, OR BUSINESS OR OTHER FINANCIAL LOSS.
For Veterinary Use Only. For In Vitro Use Only. Regulatory requirements vary by country; products may not be available in your geographic area.