Additional information

What sample material to use, how to send a sample and many more

Additional information

What kind of sample material? How to send? How long takes the testing? Here we inform you all out from the correct sampling to result interpretation! 
sample material
Choosing the right sample material is critical for a test result. On our submission forms you'll find a detailed list of usable sample material for each test. Those result from the respective test method, the clinical case and eventual symptoms (also e.g. duration of symptoms). For serological tests (e.g. determination of antibodies after vaccination) please send serum samples only. Plasma is not suited for most applications.
The following table lists some useful sample material:

list of useful sample material depending on clinical symptoms


In the following you find more information on the respective sample material.

1. Blood samples 
It's recommended to have the patient empty-stomached before blood sampling. Also the sampling should take place in a stress-free environment. To avaoid hemolysis the vein should be compressed as shortly as possible. Samples have to be taken sterilely. If using anti-coagulants the vial should be swilled around carefully.

  • Serum
Fluid and solute component of blood after fibrin clotting and separation of cellular components. Blood should remain in a vial without additives at room temperature until clotting is complete. This process can be accelerated by using clot activators. Sample should be centrifuged for 10-15 min at 1000 x g, then serum can be pipetted in another clean vial (serum amount is about 1/3 of whole blood volume). A serum sample (sterile and cell free) is quite stable, it can be cooled at 4-8° C for  1-3 days or stored at -20°C for a longer time period.

  • Plasma
Fluid and cell free component of blood after adding anticoagulants. Plasma is easier to receive than serum. Hemolysis is less likely if using the correct relation of anticoagulant and sample. Immediately vials should be swilled around carefully. After centrifugation for 5 - 10 min at 1000 x g the supernatant can be pipetted into another clean vial. Plasma is less useful for most virologic applications! Possible anticoagulants are Na/K EDTA or Heparin (Li-Heparin).

  • Whole blood sample
Blood samples without anticoagulants should not be sent to avoid hemolysis. Hemolytic samples often react toxic in tests using culture cells.

  • Blood samples with anticoagulants 
If you need to prepare leukocytes for a test, blood samples with anticoagulants are needed. It's necessary for detection of virus / viral components in blood cells (e.g. PCR). Suitable anticoagulants are Na/K EDTA or Heparin (Li-Heparin). Samples should be sent directly to the laboratory, cooling is possible but don't freeze!

  • Sample volume
For most serologic assays about 0,5 ml serum is needed. For the detection of virus in blood cells about 1 -10 ml blood with anticoagulants should be sent.

  • Toxicity (esp. serum samples)
In test methods using cell culture systems (e.g. serum neutralization assay) as a consequence of toxic effects the test cells can die and therefore which makes a test interpretation impossible. There are various reasons for toxicity of sera, e.g.  hemolysis, lipemia, icterus or drugs. Therefore it's recommended to have the patient empty-stomached before blood sampling and to avoid application of medicine if not necessary.

2. Swab samples
Pincipally sterile swabs should be used to collect liquids. The "cytobrush"-method means to rub the swab on the surface to remove cell-containing material. Swabs should be sent in sterile vials WITHOUT ANY MEDIUM, some drops of sterile sodium chloride are useful to avoid dry out. Swabs should be sent to the laboratory as fast as possible.

3. Urine samples
Sampling can made from fresh midstream urine, catheterisation or punction of the bladder. It should be sent in a closed, unbreakable tube.

4. Faeces
Faeces should be fresh and sent in the size of a pea. The container has to be closed tightly and unbreakable. A swab can be taken if pets are very small or young.

5. Cerebrospinal fluid
Cerebrospinal fluid has to be taken sterilely and without blood contaminations. The latter would make it impossible to differentiate between antibodies in the CSF and those from the blood contamination. For most assays 0.5 ml csf is sufficient (exemption: Borna RT-PCR horse, min. 10ml).

6. Punctate
Punctates have to be taken sterilely and without blood contaminations. Different punctates can be use:
  • abdominal punctate/ ascites
  • thoracal punctate
  • bladder punctate

7. Skin scrape / biopsy
Samples can be taken from various skin alterations (pocks, blisters and liquids, crusts or warts). It is useful to collect material at the border to healthy skin. Samples should be sent moisturized with sterile sodium chloride.
   Warts/sarcoids for vaccine preparation: We need about 1 g material per animal for virological tests and vaccine production.

8. Organ samples
Organ samples from freshly died or euthanized animals can be used for virological examinations after gross section. Appropriate sample material comes from affected organic systems and can have visible alterations. Samples should be in the size of a hazelnut and sent in sterile, tightly closed containers. To minimize autolysis samples should be sent as fast as possible, freezing is not recommended.
Subsmission of reports
Results are usually transmitted to the sending veterinarian directly after test analysis. Result transmission to the owner or third persons is only possible if requested by the veterinarian. For the interpretation of test results do not hesitate to contact our veterinarians for advice.

Results are communicated in written form (by email or fax). Information about test results by phone is generally restricted to the sending veterinarian, only our veterinarians are authorized to discuss test results with you. Rabies test certificates for travel purposes are printed, signed, stamped and sent by post. Sending a PDF report (b/w) in advance by email or fax is possible. 
Sample shipment
Independent of the sample material, sample should arrive in the laboratory as fast (and therefore as fresh) as possible. It is useful to send them the same day as they have been taken. If this is not possible, the correct storage of samples is mandatory. Serum samples can be cooled or even frozen for quite a long time. Whole blood or EDTA blood samples should not be frozen. If you want infectious virus to be detected in the respective sample the material should also not be frozen and thawn mor than once. Otherwise a virus can become inactivated and the test would be (false) negative. If you have older or frozen material we recommend to send it frozen (e.g. on dry ice) and to order a PCR test since DNA / RNA is more likely to be detected in this material.
To avoid damage of a sample it should be send in unbreakable (plastic) vials, liquids with some wadding material around the sample container.
All samples must be accompanied by a sample submission form, we provide forms for several animal species. For serum samples for FAVN test from non European countries we have an import permit (German "Einfuhrgenehmigung") to avoid problems at the customs.


Regulation for the transport of hazardous materials
This regulation controls the transportation of infectious materials (materials known or supposed to contain pathogenic germs hazardous for humans or animals) including diagnostic samples.

Packing instructions (in compliance with UN No. 3373 and packing instruction P650):
  • Package must be sufficiently resistant to impacts and pressures to avoid damage or discharge of contents
  • In addition to the sample container a packing container should be used
  • The package (sample and packing container) has to meet the requirements of an inspection

In detail:
1. Liquids:
  • The test tubes should be shut safely and leak-proof. The maximal permissive filling volume is 500 ml.
  • The sample container should be leak-proof and contain absorbing material (sufficient to absorb the entire sample amount)
  • The packing container should not be heavier than 4 kg
2. Solids:
  • The test tube should be dust-proof, maximal filling 500ml
  • The sample container should be leak-proof, if there is more than one sample, please avoid close contact
  • The packing container should not be heavier than 4 kg

Please note:
The dispatcher is responsible for the transported materials (liable if there is damage due to insufficient packing).

Sampling
Please keep in mind:
  • The sample should be taken under clean (sterile) conditions and be transferred immediately to a leak-proof transport container. It should be shipped as fast as possible (delicate samples may require refrigeration until/during transport).
  • For each individual animal a separate submission form should be completed including animal`s ID, vet`s and owner`s name, details about sample material, date of sampling, medical history and requested tests.
  • The sample container has to be labelled unequivocally so that each sample can be assigned to the submission form and mixing up is not possible.
Arrival of samples in our laboratory
In general samples are only accepted from veterinary surgeons, commercial laboratories, veterinary investigation and administration offices ("Veterinäruntersuchungsämter") as well as from institutes of veterinary faculties. Samples can be dispatched by mailing route or by courier service. We directly take care of arriving samples during our opening hours (Mon. - Fri., 9 a.m. - 4:30 p.m.).

Immediately after the arrival samples are unpacked and coded. The analysis of the samples takes place as fast as possible. Sample containers have to be labelled unequivocally, otherwise we will have to contact the sender. Generally, requested analyses will be performed. If there are any ambiguities concerning the requested analysis or the sample material, the quality or the quantity of the sample we will consult you immediately. We will also check if the sample material is suitable for the requested test. Please use our submission forms to include all information we need for sample processing.
Duration of the tests
All tests are performed as fast as possible. The processing time is indicated for each analysis. In exceptional cases (holidays, repeating of tests) analyses may take more time, in these cases we apologize for any inconvenience. You can always contact us by phone or email to ask about the status of your samples.
Interpretation of results
For specific questions about the interpretation of results you may contact our scientific staff who will be glad to help you. In the following you will find some general information regarding the interpretation of the results:

"negative" results
As a general rule negative results do not exclude a virus infection.

Possible reasons for false negative results:
  • Choice of samples (unsuitable samples, improper extraction time,...)
  • Sampling (unsterile samples, unsuitable culture medium,...)
  • Mailing of samples (excess of transport time,...)
  • Limitations of diagnostic methods (sensitivity, test errors)

"positive" results
A positive test result always has to be interpreted in the context with the anamnesis and clinical signs.

Possible reasons for false positive results:
  • Subclinical infections,
  • Ubiquitous germs,
  • Antibody titre caused by vaccination or maternal antibodies

Interpretation of serological results:
  • Paired serum samples taken at an interval of 2 to 4 weeks will in most cases allow the identification of an acute or subacute infection. A significant rise in antibody titres (sero conversion) is indicative in this respect.
  • Discrimination between antibodies due to vaccination or due to infection with a field virus strain will not be possible in most cases. For this reason it is essential to determine the animal's vaccination status before sampling.
  • For most virus-induced diseases thresholds for protective immunity by antibody titres are not defined. This information can only be obtained by animal experiments ("challenge-tests"). The interpretation of the test result generally only shows whether the animal has responded to the vaccination with a low, moderate, high antibody titre or not at all. Exceptions to this rule are rabies and canine parvovirus and other vaccine components where thresholds have been defined (by StIKo Vet).
  • Regarding FAVN test: titres between the lower test limit (0,03 IU/ml) and the internationally accepeted protective titre of 0,5 IU/ml are indicated "positive". Here is to say that all titers above 0,03 IU/ml have to be indicated "positive" in terms of a general detectability of antibodies. So, all detectable titers are indicated "positive" but only titers equal or above 0,5 IU/ml are being considered protective against rabies (and therefore allow a pet to enter European Union). To avoid confusions interpreting our results, we added the following explanation to our reports:
    „Antibodies against rabies virus were detectable (positive). The titre lies below 0,5 IU/ml and therefore no conclusion can be drawn with regard to the protection of the animal tested. An antibody titre of 0,5 IU/ml or above after vaccination indicates protection against rabies according to the guidelines of the WHO/OIE.“
Fees
We can send a price list to sending veterinarians on request, prices are independent of the invoiced party (veterinarian or owner). In general invoices are sent 1 week after the result has been transmitted. If the owner has accepted the meeting of the costs by signing on the sample submission form we can also sent invoices accordingly. Discounts for sending in bulk samples have to be arranged in advance.

For regular shipments we offer direct debit (German bank accounts only), the respective form can be downloaded here.

Payments from foreign countries (rabies antibody test) can be made in cash, by bank transfer or PayPal. Please always indicate our invoice number!
Complaints
If any problems, questions or complaints about procedures, results etc (e.g. rabies certificates) occur, please contact our laboratory, we will immediately try to solve the problem. You can contact us by phone, e-mail or fax. 
Please note: If a rabies certificate needs to be corrected, the original certificate has to be sent back before we can dispatch the modified certificate.
If there are any questions about invoices, please contact our accountancy Mon to Thu from 9am until 2pm (Mrs Castro-Alvarez, phone: +49  641 9938352).