Molecular Biology Sample Submission Hints
Sample Collection for Avian Influenza (AI) or Newcastle Disease Virus (NDV) Real Time (RT) Polymerase Chain Reaction (PCR) tests:
- The preferred specimens for the real-time RT-PCR assay are tracheal or oropharyngeal swabs from gallinaceous birds such as chickens, turkeys, guinea fowl, pheasants, quail, etc. For other birds, such as ducks, geese, etc., cloacal swabs are preferred
- The RT-PCR has not yet been validated for use in testing pet (caged) birds so the sample site at which the virus can be detected is not established. For this reason, in non-poultry or wild waterfowl, both oropharyngeal and cloacal areas will need to be swabbed and the samples kept separate. For non-export testing PCR will be followed up with Virus Isolation.
- Polyester tipped metal or plastic sticks (synthetic material on a synthetic shaft) are the preferred type of swabs. Wooden sticks contain inhibitors that affect the assay. Do not leave wooden shafted swabs in the medium.
- Swabs must be placed in brain-heart-infusion broth (BHI) tubes. Both swabs and BHI tubes information can be obtained suppliers such as VWR [catalog number 90001-533 (x100)] www.VWR.com, 1-800-932-5000]. Please note that the BHI broth does not have antibiotics added and the volume needs to be between 2 and 2.5ml so you will need to pour half of the volume from these 5ml tubes into a sterile vessel in order to have the right volume for the test. Swish swabs in the BHI for 10 seconds, wring them out against the edge of the tube, and discard them before sealing the tube.
- Label tubes to clearly coordinate them with the submission form. Please note that labeling on glass with a sharpie marker is not the optimal labeling method as the ice and handling can scratch off the labels. Sample identification labeling should be done on tape over glass or on a frosted area on the glass.
- Swabs can be pooled (only if also allowed by the country or state you are shipping to) as follows:
- Tracheal/oropharyngeal swabs: 5 birds/tube
- Cloacal swabs: 5 birds/tube
- DO NOT POOL cloacal and tracheal/oropharyngeal swabs together.
- Swabs from different avian species must be collected and pooled separately
- Wildlife avian influenza surveillance samples cannot be pooled
- The specimens should be kept chilled (wet ice or cold gel packs) and shipped over- night in a cooler. If samples can not be sent until the following day, keep them refrigerated, do not freeze. Samples must be tested within 4 days of collection and received cold for results to be valid.
Last Updated: December 12, 2012
