USDA Highly Pathogenic Avian Influenza Diagnostic Training Course

The Institute for International Cooperation in Animal Biologics  (IICAB) hosted five “USDA Highly Pathogenic Avian Influenza Diagnostic Training Courses”  in 2006-2007 that were funded by the USDA.  The purpose was to teach diagnosticians from laboratories throughout the world how to diagnose avian influenza and to build a network of colleagues in  this field.  In 2005, the IICAB hosted a similar course, funded by the USDA Agriculture Research Service and focused on diagnosing avian influenza and exotic Newcastle disease.  The courses were taught by scientists from the USDA APHIS National Veterinary Services Laboratories and the USDA ARS Southeast Poultry Research Laboratory. View attendee list and photographs here!

The five day course has been recorded and includes lectures on avian influenza and Newcastle disease viruses, types of diagnostic tests, molecular diagnostics, agar gel immunodiffusion, virus characterization, and sample collection, preservation and shipping.  Detailed information on laboratory tests are shown along with printable resources to help viewers follow each step in the procedures.  Additional printable resources on avian influenza are included as well.  The materials on the website are also available on DVD from the USDA or by contacting the Institute for International Cooperation in Animal Biologics at iicab@iastate.edu.   

Overview of Avian Influenza
Dr. David Suarez

Duration: 49 min.
[View Video]
Slide Presentation: [PPT] [PDF]

   
Overview of Newcastle Disease Virus
Mr. Dennis Senne

Duration: 58 min.
[View Video]
Slide Presentation: [PPT] [PDF]
   

Laboratory Orientation and Safety
Mr. Dennis Senne

Duration: 26 min.
[View Video]
Slide Presentation: [PPT] [PDF]
Biosafety in Microbiological and Biomedical Laboratories (BMBL) 4th Edition: [HTM]

   

Laboratory Diagnosis of Avian Influenza and Newcastle Disease
Mr. Dennis Senne

Duration: 54 min.
[View Video]
Slide Presentation: [PPT] [PDF]

   

Application of Real Time RT-PCR to Detect Avian Influenza Virus
Dr. David Suarez

Duration: 56 min.
[View Video]
Slide Presentation: [PPT] [PDF]

   

Characterization and Pathotyping of AIV and APMV-1
Mrs. Jan Pedersen

Duration: 35 min.
[View Video]
Slide Presentation: [PPT] [PDF]

   

Recommended Procedures for Sample Collection, Preservation and Shipping
Mr. Dennis Senne

Duration: 34 min.
[View Video]
Slide Presentation: [PPT] [PDF]

   

Avian Influenza Lab Demonstrations: All
The lab demonstrations are accompanied by the following reference guide which can be used to follow along with the video:

Lab Demonstration Reference Guide (PDF 16.4MB) 

All Lab Demonstrations (listed below)
USDA


Duration: 37 min.
[View Video]
   

Avian Influenza Lab Demonstrations: Virus Isolation
The demonstrations listed below are accompanied by the following reference material:

Virus Isolation and Specimen Processing: [PPT] [PDF

Biological Safety Cabinets
USDA


Duration: 1 min. 19 sec.
[View Video]
   
Tracheal Swabs
USDA


Duration: 45 sec.
[View Video]
   

Oropharyngeal Swabs
USDA


Duration: 30 sec.
[View Video]

   
Cloacal Swabs
USDA


Duration: 37 sec.
[View Video]
   
Tissue Samples
USDA


Duration: 2 min. 7 sec.
[View Video]
   
Candling Eggs
USDA


Duration: 1 min. 11 sec.
[View Video]
   

Egg Inoculation
USDA


Duration: 1 min. 3 sec.
[View Video]

   
AAF Collection (Dead Embryos)
USDA


Duration: 1 min. 28 sec.
[View Video]
   
AAF Collection (Live Embryos)
USDA


Duration: 1 min. 30 sec.
[View Video]
   

Avian Influenza Lab Demonstrations: HA and HI
The demonstrations listed below are accompanied by the following reference material:

Detection and Identification of Hemagglutinating Isolates: [PPT] [PDF]

USDA NVSL Testing Protocol

HI Test for Subtype Identification of Influenza A Virus Antibody: [PDF]
HI Test to Detect Serum Antibodies to Avian Paramyxoviruses: [PDF]
HA and HI Tests for Avian Paramyxovirus (APMV) Identification: [PDF]

USDA NVSL Standard Operating Procedure

Preparation of Erythrocytes for HA and HI Tests: [PDF]
Removal of Natural Agglutinins from Avian Serums: [PDF]

HA Test
USDA


Duration: 3 min. 57 sec.
[View Video]
   
HA Test Results
USDA


Duration: 57 sec.
[View Video]
   

HA Back Titration
USDA


Duration: 2 min. 5 sec.
[View Video]

   
HI Test
USDA


Duration: 3 min. 37 sec.
[View Video]
   

Avian Influenza Lab Demonstrations: ELISA
The demonstration listed below is accompanied by the following reference material:

ELISA: [PPT] [PDF]
Laboratory Handout: [PDF]

IDEXX ELISA
USDA


Duration: 3 min. 18 sec.
[View Video]


   

Avian Influenza Lab Demonstrations: Antigen Detection
The demonstation listed below is accompanied by the following reference material:

AIV Antigen Detection: [PPT] [PDF]
Laboratory Handout: [PDF]

Antigen Detection
USDA


Duration: 1 min. 31 sec.
[View Video]


   

Avian Influenza Lab Demonstrations: rRT-PCR
The demonstations listed below are accompanied by the following reference material:

Recommended Procedures for the Extration of RNA: [PPT] [PDF]
Amplification and Detection of Nucleic Acid by the Real-Time RT-PCR Procedure: [PPT] [PDF]
Interpretation of Results: [PPT] [PDF]
rRT-PCR Lab Handouts: [PDF]

NVSL Testing Protocol

Real-Time RT-PCR for Detection of Virulent Newcastle Disease Virus in Clinical Samples: [PDF]
Real-Time RT-PCR for Detection of Avian Influenza Virus and Identification of H5 and H7 subtypes in Clinical Samples: [PDF]

Ambion® Magnetic Bead Extraction
USDA


Duration: 3 min. 47 sec.
[View Video]

   

Qiagen® RNeasy® Mini Columns
USDA


Duration: 4 min. 6 sec.
[View Video]

   

Master Mix Preparation
USDA


Duration: 3 min. 22 sec.
[View Video]

Other Resources Distributed to Course Participants

Highly Pathogenic Avian Influenza
OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals 2004 (Chapter 2.1.14): [PDF]
OIE Terrestrial Animal Health Code 2004 (Chapter 2.7.12): [PDF]

Newcastle Disease
OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals 2004 (Chapter 2.1.15): [PDF]
OIE Terrestrial Animal Health Code 2004 (Chapter 2.7.13): [PDF]