a unique bvdv screening and identification protocol
TRANSCRIPT
A Unique BVDV Screening and Identification Protocol.
BVD 2006 CONFERENCE
Why is There BVDV in the USA?Why is There BVDV in the USA?
• Industry Resistance
• Lack of Individual Identification
• Lack of Good Screening Tools
• Industry Resistance
• Lack of Individual Identification
• Lack of Good Screening Tools
A Perfect PI BVDV Screening TestA Perfect PI BVDV Screening Test
• 100% Sensitivity– Moderate Specificity is OK– Acute vs Persistant Infection
• High Throughput– Sampling– Assay Technology
• Low Cost
• 100% Sensitivity– Moderate Specificity is OK– Acute vs Persistant Infection
• High Throughput– Sampling– Assay Technology
• Low Cost
Current BVD Diagnostic Tests Current BVD Diagnostic Tests
VIVI IHCIHC ELISAELISA
Single Single Sample Sample
PCRPCR
GoodGood GoodGood GoodGood GoodGood SamplingSampling
PoorPoor PoorPoor GoodGood PoorPoor ThroughputThroughput
PoorPoor PoorPoor FairFair FairFair Turn AroundTurn Around
GoodGood GoodGood GoodGood GoodGood SensitivitySensitivity
FairFair FairFair GoodGood PoorPoor TechnologyTechnology
PoorPoor PoorPoor FairFair PoorPoor CostCost
Reverse-Transcriptase Polymerase Chain Reaction on Pooled Samples to Detect Bovine Viral Diarrhea Virus by Using Fresh Ear-Notch-Sample Supernatants
James A. Kennedy, Robert G. Mortimer, Barbara Powers
J. Veterinary Diagnostic Investigations (2006)
Reverse-Transcriptase Polymerase Chain Reaction on Pooled Samples to Detect Bovine Viral Diarrhea Virus by Using Fresh Ear-Notch-Sample Supernatants
James A. Kennedy, Robert G. Mortimer, Barbara Powers
J. Veterinary Diagnostic Investigations (2006)
PCR Analysis of Pooled SamplesPCR Analysis of Pooled Samples
• Jim Kennedy’s Screening Protocol
– Screen 100 Pooled Samples per PCR Reaction
– Sort and Confirm Positives by ELISA
– Demonstrated Sensitivity and Specificity
• Jim Kennedy’s Screening Protocol
– Screen 100 Pooled Samples per PCR Reaction
– Sort and Confirm Positives by ELISA
– Demonstrated Sensitivity and Specificity
Detection of BVDV RNA by RT-PCRDetection of BVDV RNA by RT-PCR
MWSTD
1x 0.1x
0.01
x
600 kb
PCR Analysis of Pooled SamplesPCR Analysis of Pooled Samples
• Issues
– Technically Challenging
– Throughput is Moderate
– Polymerase Inhibitors
• Issues
– Technically Challenging
– Throughput is Moderate
– Polymerase Inhibitors
PCR Analysis of Pooled SamplesPCR Analysis of Pooled Samples
• Potential Solutions
– Real Time Analysis
– Optimize RNA Preparation
– Automation
• Potential Solutions
– Real Time Analysis
– Optimize RNA Preparation
– Automation
Front End ObstaclesFront End Obstacles
• Identification
• Sampling
• Shipping
• Identification
• Sampling
• Shipping
Stores all information on the tag
Ranch IDRanch IDBirth dateBirth dateISO #ISO #Visual IDVisual IDTreatment dateTreatment date
Sampling Sampling
A Unique BVDV Screening and Identification Protocol.
A Unique BVDV Screening and Identification Protocol.
• Sampling• Shipping• Write Tag• Screening and Diagnosis• Consulting
• Sampling• Shipping• Write Tag• Screening and Diagnosis• Consulting
Special Thanks to James Kennedy, DVM
Special Thanks to James Kennedy, DVM
BVD 2006 CONFERENCE