Regional Diagnostic Virology Services (RDVS) at AVC Diagnostic Services

General Information

The Regional Diagnostic Virology Services (RDVS) is a major component of AVC Diagnostic Services at the Atlantic Veterinary College. It was established and opened for service in 1988, two years after the Atlantic Veterinary College started. Prior to this, the diagnostic virology services for the Atlantic Region were being done by Agriculture and AgriFood Canada in Sackville, New Brunswick using traditional virology test, e.g. virus isolation. It has to be emphasized that the RDVS at AVC was developed and established through the financial support of the veterinary laboratories and agriculture department of the four Atlantic Provinces (New Brunswick, Newfoundland, Nova Scotia and Prince Edward Island) through grant and a fee for service arrangement until 2013 (25 years). The provincial laboratories remain strong clients of RDVS at AVC utilizing another agreement since 2013.

The RDVS initially offered diagnostic virology services for food animals (bovine, porcine, avian, caprine, ovine), equine and furbearing animals in the region as a part of the regional diagnostic agreement. It has since expanded to include companion animals and wildlife for practitioners and other clients in the region.

The RDVS offers a wide range of diagnostic virology tests for mammalian species from traditional methods such as virus isolation using cell culture to the established rapid tests such as negative staining electron microscopy, enzyme linked immunosorbent assay (ELISA), fluorescent antibody test (FAT) and molecular diagnostic test such as polymerase chain reaction (PCR), reverse-transcriptase PCR (RT-PCR), nested PCR (nPCR) and real time PCR (rt PCR). The request for traditional tests such as virus isolation (which has a long turnaround time, laborious and expensive) has gradually decreased in numbers and has been replaced by molecular diagnostic tests, FAT, and ELISA. However, virus isolation for mammalian species is still available for special cases (undiagnosed disease outbreaks, legal issues, etc.) for clients (pathologists and practitioners) in the region.

In 1993, the RDVS developed and established virology services for aquatic species initially in the Atlantic Region and has since expanded nationally and globally. The most popular test that was offered for this species was virus isolation using one to several cell lines (up to 5-7). The molecular diagnostic tests such as PCR, RT-PCR, nPCR and rt-PCR for specific virus(es) were also developed and established.

The RDVS is accredited by Canadian Food Inspection Agency (CFIA) for two tests-Equine Infectious Anemia and Enzootic Bovine Leukosis. In 2014, the RDVS laboratory achieved ISO/IEC 17025:2015 certification by the Standard Council of Canada.

The RDVS is a member of the Canadian Animal Health Surveillance Network (CAHSN) and actively performs tests mandated by CFIA and CAHSN.

Requirements for Sample Submission

All samples for virology must be submitted through a licensed veterinarian or provincial laboratories. The submitter becomes responsible for test(s) requested and fees for each submission.

Samples received for testing become the property of AVC Diagnostic Services. The RDVS Laboratory does not accept sample(s) submitted by the owner.

General Virology Submission Forms

There were three virology submission forms which can be downloaded and printed from the computer. They are as follows:

  1. Food and Fur Bearing Animal Virology Submission Form http://files.upei.ca/avc/diagnosticservices/QA-F-101B-01_MC.pdf
  2. Multiple Animal Virology Submission Form (for multiple tests and multiple samples) http://files.upei.ca/avc/diagnosticservices/QA-F-101D-01_MC.pdf
  3. Companion Animal Virology Submission Form. http://files.upei.ca/avc/diagnosticservices/QA-F-101A-01_MC.pdf

Please fill out and complete the appropriate form. Written history should briefly be filled out in appropriate place in the form. Please use back of the form, if needed.

CFIA Regulatory Forms

The RDVS Laboratory is accredited to perform Equine Infectious Anemia (EIA) ELISA and Enzootic Bovine Leukosis (EBL) ELISA by CFIA. Only accredited veterinarians can collect and send samples for these tests.

The following official forms are required:

  1. EIA ELISA (CFIA Form No. 3937)
  2. BLV ELISA (CFIA Form No. 3841)

The accredited veterinarians can request these forms from their respective CFIA District Offices.

Please complete all necessary boxes in the form. Please ensure that the information including owner’s information, animal location, reason for test and description of the horse is completed. Also, please ensure that the names of the accredited veterinarian, their signature, as well as the sampling date are included.

Specimen Selection and Collection

Clinical virology samples should be collected as soon as possible following the onset of illness. Please refer to the AVC Diagnostic Services Website where you will find a link to the ”Detailed Guide in the Selection and Collection of Virology Specimen” http://files.upei.ca/avc/diagnosticservices/AVCDS_DetailedGuideSelectionCollectionVirologySpecime ns.pdf . The table shows the available tests for specific virus(es), the recommended specimens for each test and information on the collection procedure and transport devices that can be used.

All specimens must be properly identified and labelled. The samples must be identified with Animal ID or laboratory or hospital reference number which is also present on the sample submission form.

Virus Isolation:

VI samples must be labelled with the origin of the specimens (e.g. nasal swabs, name of tissue or name of pooled tissues, etc.). The specimens for virus isolation should be collected aseptically and should be refrigerated promptly. Materials can be collected (using special swabs and special viral transport medium provided by the RDVS) from nasal sinuses, conjunctival surfaces or tissues. If samples cannot be sent within 24 hours (e.g. weekend), freeze at -700C to -800C. If this type of freezer is not available, store at refrigeration temperature (40C). Do not freeze specimens for virus isolation at -200C or regular freezer temperature.

Negative Staining Electron Microscopy:

The tracheal or bronchoalveolar washings should be placed in leak-proof container. For EM, 5-10 grams or 5-10 ml of feces or intestinal contents is required. If possible, feces should be collected directly from the animal. The intestine containing fluid materials should be tied on both ends before placing in a leak-proof container.

Molecular Diagnostic Test:

For molecular diagnostic tests, please use special swab and special viral transport medium (provided by the RDVS) for feces, intestinal content, nasal sinuses, conjunctival and laryngopharyngeal specimens. When submitting tissues, please collect as aseptically as possible and place it in a plastic bag before putting it in a leak-proof container.

Serology:

A minimum of 1.0 ml of serum is needed for a single test. For more than one test, a minimum of 2.0 ml is needed. Collect whole blood in red-top vacutainers. Please note that 10.0 ml of blood can generate 3-5 ml serum. Allow the samples to clot at room temperature. Release the clot using a sterile applicator stick, spin down and transfer the serum supernatant to a sterile tube. If a centrifuge is not available, remove the clot and set aside the serum.

For serodiagnosis, paired serum samples are required. The first sample is collected at the onset of illness and the convalescent is collected 2-3 weeks later. Acute sample(s) should be stored at -20C while waiting for the convalescent samples.

There are situations where the submitting of a single samples is necessary. Please specify the reason(s) in your submission form, e.g. sale, animal has died, determination of vaccination titer, etc.

Paired serum samples must be labelled as acute (A) or convalescent (C).

Fluorescent Antibody Test:

The specimens should be refrigerated or frozen very soon after collection. Place these in sterile plastic bags or leak-proof containers. The name of the organ(s) must be marked in the container. Around 3 to 5 cm tissue(s) should be collected. For smaller animals and birds, include the whole organs.

Special Virology Swabs and Viral Transport Medium

These special supplies can be requested from RDVS Laboratory. The cost is $3.00 per unit. If requested, the RDVS can send these to appropriate clinic or laboratory using courier collect.

Packing and Shipment of Specimens for Virology Testing

Leak-proof primary containers with feces (reinforced with tape, if needed), leak-proof primary containers with tissues, tubes containing viral transport medium and tube(s) with blood or serum should be placed in a suitable secondary container such as sealed plastic bags after disinfecting the outside of the primary container. Multiple tissues can be placed in the same bag, as long as they are from the same animal and there is sufficient absorbent material to absorb in case of leaks. Disinfect the outside of the secondary leak-proof container before final packaging.

The specimen(s) should be placed in a refrigerated container (Styrofoam box) with ice packs and should be surrounded with sufficient absorbent material to absorb in case of leaks.