Chapter 3 - Testing
3.2 Serologic Testing
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This part of the Accredited Veterinarian's Manual will provide the following information necessary for serologic testing:
- collection, material and supplies
- preparation and packaging of serum samples
- laboratories
- submission forms
- paper submissions
- digital submissions with a Canadian Food Inspection Agency-approved digital Equine Infectious Anemia (EIA) certification system
- special procedure for submission to the CFIA laboratories, applicable to samples from bovine tested in isolation (IAI notification), from sheep and goats, wild boar or cervids for export to Mexico and bovines tested under the Canadian Health Accredited Herd – Enzootic Bovine Leukosis (EBL-CHAH) program
- shipping samples
- criteria for rejection of samples and submission forms in CFIA animal health laboratories
- test results
Objectives
Section 2 of the Health of Animals Regulations interprets "test" as including:
- the collection of body tissue or fluid from an animal (for example, serum)
- the injection of an animal for the purpose of determining that animal's freedom from infection with disease (for example, tuberculin testing)
The reliability of any diagnostic laboratory procedure depends directly on the type of specimens or samples received and on their condition. The laboratory must receive a sample of good quality for diagnostic purposes.
Collection, materials and supplies
It is extremely important that animals are identified correctly. Animals which are tested must be uniquely identified using a approved indicator for the species being tested. Where no approved indicator exists for species tested, refer to Module 2.1 Identification of Livestock or Module 2.2 Identification of horses for more information.
Unless otherwise indicated, a minimum of a 1 ml sample of serum is required for each test.
- if using serum separator tubes (SSTs), ensure that they are centrifuged at a high enough g-force (1100-1300 relative centrifugal force) to properly seat the separating gel.
- After centrifugation, decant the serum from the SST into a new red top tube before shipping to the laboratory.
- the centrifuged SST tube alone will NOT prevent hemolysis and leakage of blood cell components back across the plug into the serum fraction.
A separate sample must be submitted for each diagnostic test required even if various tests are requested and are done at the same laboratory. Laboratories prefer separate serum samples for each test in order to maintain sample integrity. For example, if brucellosis and leptospirosis tests are requested, 2 samples are required.
For clotted blood samples:
- use silicone coated tubes (red stopper), which should be centrifuged before shipping and the clot removed
- serum should be kept refrigerated until shipment
- use disposable needles
- use a waterproof marker to number the vials
Small (3 ml) vials with or without internal thread closures should be avoided as these are prone to leakage and may cause technical problems for the laboratory.
The costs associated with material and supplies must be paid by the accredited veterinarian.
Preparation and packaging of serum samples
Each vial must be marked indelibly with the number corresponding to the one entered on the submission form in the column under Vial No. Label the vial, not the stopper.
Paired samples must be identified with the same vial number for the same animal.
- the acute sample (the oldest one) will be identified A, and the convalescent sample (the most recent) will be identified C (for example: 001A, 001C, 002A 002C, 003A, 003C, etc.)
Accredited veterinarians must make sure that the packaging and shipping procedures used complies with regulations concerning the transportation of such samples.
- the majority of samples collected under the Accredited Veterinarian Program will be classified as "Exempt Animal Specimens" under the Transportation of Dangerous Goods Regulations
- Exempt Animal Specimens are defined as "Animal specimens not believed to contain infectious substances (for example: samples taken for surveillance or export purposes) or specimens where the pathogens has been neutralized/inactivated"
- for samples not meeting this definition, consult with you District Veterinarian as to the proper classification and shipping procedures required
- refer to the figure below for the standards required when shipping Exempt Animal Specimen
Outer package markings checklist
- name, address, phone number of the shipper and consignee
- the words – "Exempt Animal Specimen" on the packaging
Statements on Shipper's Waybill
- statement – Exempt Animal Specimen
Note: if multiple fragile primary receptacles are placed in a single secondary packaging, they must either be individually wrapped or separated to prevent contact between containers.
Vials should be packed in boxes designed for this purpose in a manner to prevent breakage (for example, a New York box or styrofoam vial shipper).
- the use of an appropriate container is a requirement when submitting samples to a laboratory
- in addition to a legal responsibility, a submitter has a moral obligation to ensure the safety of transportation industry workers and laboratory staff handling the specimen
The laboratory submission forms should be placed in an envelope and attached to the outside of the protective containers.
Laboratories
Most testing, mandated by the Health of Animals Act and regulations or conducted in response to a specific request by an importing country, is conducted in a CFIA laboratory or a CFIA-approved laboratory, unless specified otherwise.
Certain private laboratories are specifically approved by the CFIA to conduct official tests for Equine Infectious Anemia (EIA) and Brucellosis submitted by accredited veterinarians.
- in such cases the accredited veterinarian manual will specify that the test may be performed in a private or authorized laboratory that meets the requirement of the Policy on the Use of External Laboratories
- the accredited veterinarian must verify with the CFIA district office that the laboratory meets the specified requirements before sending a sample
The accredited veterinarian is responsible for the laboratory fees.
- applicable fees for test submitted to CFIA laboratories are available in Module 4.2 CFIA Fees
All specimens sent to a CFIA or CFIA-approved laboratory must be submitted with a CFIA-approved submission form for the official test being requested.
- please contact the CFIA-approved laboratory directly to confirm what documentation or notification is required when using a CFIA-approved digital EIA certification system for sample submission. (Contact information available in Module 3.3)
Paper submission forms
Submission forms for samples sent to CFIA laboratories are available at the CFIA district office.
- for samples submitted to a CFIA laboratory, use Form CFIA/ACIA 5473 – Animal Health Import, Export and Artificial Insemination Specimen Submission
- instructions for submission of samples are printed on that form
- a laboratory notification number, which is available from a CFIA district veterinarian, must be entered in the "Reason for Test" section of Form CFIA/ACIA 5473
- ensure the correct notification number for the samples being tested is used
- for each test requested, a serum sample and a copy of the appropriate completed submission form must be sent to the laboratory
- a copy of the submission form must be kept on file and a copy must be sent to the CFIA district office when the sample is submitted to a CFIA laboratory
To submit brucellosis samples to a CFIA-approved laboratory, use Form CFIA/ACIA 5473 as well.
- when samples are tested for export purpose, a notification number is not required on Form CFIA/ACIA 5473, but the destination should be identified as "USA or Mexico"
it is acceptable to complete the first 2 pages of Form CFIA/ACIA 5473 in full and mark the third page with "See attached list"
The list can be a spreadsheet that records the animals' identification, description and the vial numbers, and should identify "form reference #_______________" and be signed.
- the form reference number is the number which appears in the bottom centre of all 3 pages of Form CFIA/ACIA 5473 and must begin with the current calendar year
- it should be noted that each CFIA Form 5473 bears a unique reference number and can only be used once
- Do not make photocopies of these forms for later use
Note: for brucellosis samples collected for purposes other than export or the artificial insemination program (for example pre movement of zoological species), please consult with the local CFIA District Office for proper submission procedures.
To send EBL-CHAH samples to a CFIA laboratory, use Form CFIA/ACIA 3841 – Canada Health Accredited Herd – Serum Test Report.
- an electronic template for submission of EBL-CHAH samples must be used in lieu of filling out the Animal Information Table of the CFIA 3841 (see below)
For accredited veterinarians working for clinics who have applied for and been granted a "digital exemption" (refer to Module 8.4 Equine infectious Anemia), EIA samples can be sent to a CFIA-approved laboratory using Form CFIA/ACIA 3937 – Equine Infectious Anemia (EIA) Serum Test Report and Certificate.
Forms should be typed or printed and all copies must be legible.
- all information should be recorded on the form including ear tag numbers, tattoos or other types of identification
- the type of test being requested and the date and location where the specimens were collected should be recorded
- accredited veterinarians must include their name, address (including postal code) and telephone number, and must sign at the bottom of the form
- the submitter code must also be indicated, as well as the identification code of the CFIA district office in which the animals tested are located
It is the responsibility of the accredited veterinarian to provide all information required.
- laboratories may refuse to conduct analysis of samples submitted with forms that are not fully complete
- please refer to a CFIA district veterinarian for examples of completed forms and for instructions on the distribution of forms for samples being sent to CFIA laboratories
Digital submissions using a CFIA-approved digital EIA certification system
For accredited veterinarians working for clinics who have not been granted a "digital exemption", a CFIA-approved digital EIA certification system must be used to submit samples for EIA testing to a CFIA-approved laboratory.
- data fields of the electronic system are based on Form CFIA/ACIA 3937
- for certification system provider contact information, please refer to the List of CFIA-approved Electronic EIA Certification Systems in Module 8.4 Equine infectious Anemia
The accredited veterinarian must:
- register with the service provider and receive the necessary training
- send samples to a CFIA-approved laboratory which is registered with the same provider (laboratory contact information available in Module 3.3)
For more details on the required information and digital photographs for identification, please refer to Module 8.4 Equine Infectious Anemia.
Accredited veterinarians are responsible for the security and proper use of their digital signature and shall take reasonable care to prevent its misuse.
only accredited veterinarians are authorized to sign documents using their password protected digital signature
Other clinic staff does not have the authority to be part of the signing process. Utilization of your digital signature by anyone other than yourself would be considered a breach of your agreement as an accredited veterinarian and would render the form invalid.
- please note that provisions of article 10 from your Accredited Veterinarian Agreement also apply to the digital submission of documents through an digital system
Please contact the CFIA-approved laboratory directly to confirm what documentation or notification is required when using a CFIA-approved digital EIA certification system for sample submission. (Contact information available in Module 3.3).
Final CFIA-approved digital EIA test certificates are recognized as official EIA test results and can be accessed online in real-time by the owner (optional), the accredited veterinarian, the CFIA-approved laboratory and CFIA.
- Black and white or colour printed copies of the electronic EIA test certificate are both acceptable and can be used for domestic use within Canada and as proof of a negative EIA test result for export purposes
For additional information regarding the use of an approved digital EIA certification system, please contact the CFIA district office.
Special procedure for submission to CFIA laboratories
This section is applicable to samples from bovine tested in isolation (IAI notification), from sheep and goats, wild boar or cervids for export to Mexico and from bovine tested under the EBL-CHAH program
When an accredited veterinarian needs to send samples to be analyzed in CFIA laboratories from bovine under isolation notification (IAI), sheep and goats, wild boar or cervids for exportation to Mexico or bovines under the EBL-CHAH program, an Excel spreadsheet must be completed and forwarded electronically in addition to the paper form submission (CFIA Form 5473 or CFIA Form 3841) that will be attached to the samples to be sent.
- The template for each activity listed above is available through your local CFIA district office
The spreadsheet will be used by the CFIA laboratories to import data into the Laboratory Sample Tracking System (LSTS), preventing transcription errors and saving time, especially on larger submissions.
The template must be completed as follows:
- for each submission, a new template will need to be completed
- make sure to save it under a new name containing the reference number of the CFIA Form 5473 form it will refer to (for example 2014IEAB-0000079706-4)
- for EBL-CHAH submissions, the file name should contain the name of the herd sampled and date of testing
- the properly named Excel file must be sent to a generic email address: cfia.labtemplate-gabaritlab.acia@inspection.gc.ca
- a copy must be sent as well to the local district office where the animals were located when tested
- each email must contain only 1 attached template
- in the email subject line, provide the CFIA Form 5473 reference number or name of the EBL-CHAH herd only
- in the body of the message, you may provide info on the date the samples are shipped, if possible
no column can be deleted or added to the template and the mandatory format is to be followed
Otherwise, the importation of data at the laboratory level will fail leading to a delay in the analysis until a new template is forwarded.
- mandatory fields in the template include sample No., date sampled, sample type, animal identification (ID) (at least 1 of the 6 fields about ID is mandatory) and breed
- enter the age in column N
- if the age is unknown this column may be left blank as long as you select from the drop down menu "estimate" for column L and "not provided" for column M
- some fields do not apply for isolation testing or for export to Mexico
- They can't be deleted but may be left blank.
The generic laboratory email address is not assigned to a specific laboratory and cannot be used to submit questions or requests
- Any questions will need to be directed to the local CFIA district office
The electronic submission of the template does not preclude the requirement to send a paper copy of the submission form along with the sample.
- it is acceptable to complete the submission form in full and mark "See attached list" in the animal information table
- the list can be a copy of the electronic spreadsheet that records the vial numbers, the animal's identification, identify the "form reference #_______________" and be signed
- the reference number is the number which appears in the bottom centre of all 3 pages of Form CFIA/ACIA 5473
- to be acceptable, the attached list must show all the information for each animal on the same page (use the hide function from the spreadsheet to remove unnecessary columns), the reference number must be written on every page, and each page needs to be initialed by the accredited veterinarian
Shipping samples
Include all pertinent/required forms with the samples.
- Please contact the CFIA-approved laboratory directly to confirm what documentation or notification is required when using a CFIA-approved digital EIA certification system for sample submission. (contact information available in Module 3.3)
The shipment of samples must never be entrusted to the animal owner or exporter.
- the accredited veterinarian must be able to maintain a chain of custody for samples shipped to the laboratories
- samples to be shipped may be grouped at an intermediary location into a larger shipment provided that the samples are not opened and / or repackaged after leaving the accredited veterinarian possession
After collection, samples are refrigerated and shipped to the laboratory without delay. Use the safest and quickest means of shipment available.
Ship samples only during the week to avoid having them delayed in transit or held over at the laboratory on weekends or holidays.
- it is advised to only ship samples Mon-Wed, unless receipt at the lab can be guaranteed prior to close of business for the weekend
- pay particular attention to holiday closures
- in case of delayed shipment, serum samples for EIA may be frozen and sent at a later date
- consult Module 8.4 Equine Infectious Anemia for detailed instructions regarding storage and timing of analysis to ensure validity of results
- Samples for virus isolation tests such as Bovine viral diarrhea – Immunoperoxidase (BVD-IP) must not be frozen prior to submission, and must be protected from extremes of temperature during transit
- any factor that affects the viability of the agent in question will affect the sensitivity of the test and therefore the reliability of the test result
Vial breakage will require re-bleeding an animal and significantly prolong laboratory test times.
It is the responsibility of the accredited veterinarian to resubmit if a specimen is unfit for testing. Refer to the tables below for common causes of "unfit" samples.
The accredited veterinarian is responsible for shipping costs.
- if samples are sent by mistake to the wrong CFIA laboratory, the laboratory may as a courtesy resend it to the appropriate laboratory, if the accredited veterinarian has agreed to provide their courier account number to cover the shipping cost
Criteria for rejection of samples and submission forms in CFIA animal health laboratories
CFIA and CFIA-approved labs must adhere to quality assurance standards in the receipt and testing of all samples received. CFIA-approved labs set their own internal criteria for the rejection of samples received, and have protocols in place to notify the submitters of samples rejection and the reasons for the rejection. These criteria can be obtained directly from the specific CFIA-approved lab. For samples submitted to CFIA labs, criteria for rejection are listed in the tables below.
Reasons for rejection may be related to the sample itself or the sample submission form. Notification of rejection of samples will come to the accredited veterinarian through the CFIA District Office and will consist of the submission reference number, vial(s) number(s) of the rejected samples (if applicable) and the reason for rejection. Based on the reason for rejection, the accredited veterinarian will be required to undertake the appropriate corrective measures for the sample(s) and/or the submission form in order to complete the testing.
A – General for all sample types
No. | Criteria of rejection for samples | Result on Record of Analysis (ROA) | Comments |
---|---|---|---|
A1 | Contamination | Unfit |
|
A2 | Incorrect type of sample or container | Unfit |
|
A3 | No ID on sample's container (vial/bag) | Unfit |
|
A4 | Error in ID | Unfit |
|
A5 | Duplicate samples | Unfit |
|
A6 | Missing sample | No sample received (NSR) |
|
A7 | Extra sample | Sample is not tested |
|
A8 | Broken vial | Broken on arrival |
|
A9 | Insufficient volume/quantity | Insufficient sample |
|
A10 | Sets of samples with the same IDs in a unique box | Unfit |
|
B – Serology samples
No. | Criteria of rejection for samples | Result on Record of Analysis ROA | Comments |
---|---|---|---|
B1 | Lipaemia | Unfit |
|
B2 | Haemolysis | Unfit |
|
B3 | Fibrin clot (avian samples) | Unfit |
|
C – Bovine Spongiform Encephalopathy (BSE) samples
Note: BSE testing is not done under the Accredited Veterinarian Program, but accredited veterinarians may be involved in the collection of these sample on behalf of CFIA
No. | Criteria of rejection for samples | Result on Record of Analysis ROA | Comments |
---|---|---|---|
C1 | Damaged sample | Not tested – No sample |
|
C2 | Autolyzed sample | Not tested – Unsuitable |
|
C3 | Incorrect type of sample | Not tested – No sample |
|
D – Other specific sample types
Note: these specifications apply to samples taken for routine tests and are an excerpt from the document Animal Health Tests and Analyses performed in CFIA laboratories
No. | Criteria of rejection for samples | Result on Record of Analysis ROA | Comments |
---|---|---|---|
D1 | Avian Influenza | Unfit |
|
D2 | Avian Paramyxovirus (Newcastle Disease) | Unfit |
|
D3 | Bovine Viral Diarrhea | Unfit |
|
D4 | Campylobacter fetus | Unfit |
|
D5 | Contagious Equine Metritis | Unfit |
|
D6 | Cysticercosis | Unfit |
|
D7 | Elaphostrongylus cervi | Unfit |
|
D8 | In-vitro fertilized embryo – virus isolation | Unfit |
|
D9 | Infectious Bovine Rhinotracheitis | Unfit |
|
D10 | Mycoplasma | Unfit |
|
D11 | Salmonella | Unfit |
|
D12 | Tritrichomonas (Trichomonosis) | Unfit |
|
D13 | Turkey Rhinotracheitis (TRT) | Unfit |
|
D14 | All Histopathology tests (for example Cysticercosis, TB) | Unfit |
|
Test results
The accredited veterinarian is responsible for advising the owner of the results.
Test results reported from CFIA laboratories are generated through the Laboratory Sample Tracking System (LSTS).
- the test results will be provided to the accredited veterinarian by the local district office
Test results from CFIA-approved laboratories are usually reported directly to the submitter and the CFIA district office.
For additional information on distribution of test results when using a CFIA-approved electronic EIA certification system, please refer to Module 8.4 Equine Infectious Anemia.
Test results from private laboratories are usually reported directly to the submitter only. The accredited veterinarian will provide a copy of such test results to CFIA upon request.
Schedule testing so as to allow sufficient time to receive results.
- turn around times for samples tested are relative to the day the samples are received by the laboratory.
- Date modified: