LABORATORY FOR CLINICAL DIAGNOSTICS
Directory of tests 2019/20 2019/20 Foreword LABORATORY FOR CLINICAL DIAGNOSTICS Dear colleagues, you are now holding our catalogue, very Our fields of interest: conventional, indeed, as a paper version, First things first - taking care of the but also available as a digital one to analytics to serve your needs, to serve browse online. So many lab manuals exist. your clients, is our first and foremost Still, we believe we have put together goal. At the same time, whenever we find some useful information that is not so the existing analytics lacking something, easily at hand, a very unique manual either accuracy or speed or availability, indeed. we will look into that in detail. Research has always been part of our work to gain Our lab profile: access to innovative new methods or You will find some basic parameters tests. You can rely on us! and profiles as well as some unique parameters that are not so frequently Our service: found elsewhere but very helpful. We take From sample collection to availability of pride in establishing new tests, always results 24/7 - we are convinced that it is searching for the best possible technique, our duty to make analytics and the work the best possible parameter. Some with the lab as easy as possible. Invoicing specialties: You will find a vast range of is either by order or on a monthly basis. tests especially created to meet the needs And turnover dependent debates are in the diagnostic field of small mammals something we give regularly, no need and reptiles. And our antibiotic testing to contact us. Plus, our representatives is taylor-made to help with an optimised throughout the European countries will be therapy for all species involved. happy to help either by phone, fax or by paying a visit whenever a problem arises. Our quality: Just let us know please! Accreditation ensures that the test quality We want you to be able to concentrate on is constant and certified by external diagnostics and therapy - just leave the specialists. You as our customer can lab part to us! rely on that. Accreditation audits the qualification of the people involved as Our vision: well. You can be assured, our system In an ever-changing world there is a of constant learning is set up to make constant need for accuracy and reliability innovation possible within our lab. The for the vet when commissioning the lab. growing number of specialists, including We promise to give you the best possible diplomates in various fields and members support. of university staffs working for us makes You can rely on us! it possible for us to take an active part in congresses and seminars throughout Kind regards Europe. At the same time, we love routine diagnostics and we are happy to discuss cases with you in practice. Dr. Elisabeth Müller CEO LABOKLIN GmbH & Co KG
3 2019/20 LABOKLIN at a Glance 2019/20 Contents LABORATORY FOR CLINICAL DIAGNOSTICS LABOKLIN at a Glance Contents LABOKLIN is accredited according to DIN EN ISO/IEC 17025:2005 Foreword 4 2.1.4 Blood Donor Profiles – Small Animals 39 An overview of LABOKLINs range of services LABOKLIN at a Glance 4 2.1.5 PCR Profiles – Dog/Cat 40 Profiles and Screenings Pathology 2.1.6 Faecal Profiles – Contents 5 • Dog and cat • Histopathology Small Animals 42 • Small mammals • Immunohistology Our contacts 8 2.2 Profiles – Small Mammals, • Birds • Cytology Birds and Reptiles 43 • Reptiles • BRAF Mutation Abbreviations 11 2.2.1 Clinical Chemical • Horse • Exsudate/Transudate Profiles 43 1 Pre-analytics 13 • Ruminants • Cerebrospinal fluid 2.2.2 PCR Profiles – 1.1 Blood, plasma, serum samples 13 • New World camelids • Synovia Small Mammals, • Pig • Other aspirates 1.1.1 Preparation of the Birds and Reptiles 45 • Amphibians patient 13 2.2.3 Faecal Profiles – PCR detection 1.1.2 Which sample? 13 Small Mammals, Blood examinations • Dog 1.1.3 Factors interfering with • Allergy • Cat analysis 15 Birds and Reptiles 47 • Endocrinology • Small mammals 1.1.4 Specific features 17 2.3 Profiles/Screenings – Horse 48 • Function tests • Birds 1.2 Microbiology 17 2.3.1 Clinical Chemical • Haematology • Reptiles 1.3 Hygiene 18 Profiles 48 • Clinical chemical parameters • Horse 1.4 Histology and immuno 2.3.2 PCR Profiles – Horse 52 • Serology/Infectious diseases • Cattle histochemistry 18 2.3.3 Faecal Profiles – Horse 53 • Pig 1.4.1 Skin punches 19 2.4 Profiles/Screenings – Hereditary diseases • Amphibians 1.4.2 Cytology 19 Ruminants 54 • Dog • Fish 1.5 Polymerase chain reaction 2.4.1 Clinical Chemical • Cat • Sex determination in birds (PCR) 20 Profiles 54 • Horse • etc. 1.6 Genetic testing 20 2.4.2 Serological Profiles – • Cattle 1.7 Immune status 22 Ruminants 56 • Pig Other genetic examinations 1.8 Sample material/Shipping 2.4.3 PCR Profiles – • etc. • Breed analysis material 22 Ruminants 56 1.9 Labelling 24 • Identity and parentage 2.4.4 Faecal Profiles – 1.10 Packaging and transport 26 Hygiene • DNA profile Ruminants 57 • Hygiene examinations • Species differentiation 1.11 Reordering tests 27 2.5 Profiles/Screenings – Pig 58 • Profiles • Coat colour/coat structure 2 Profiles and Screenings 29 2.5.1 Clinical Chemical Profiles 58 2.5.2 Serological Profiles – Pig 59 Microbiology and parasitology Preanalytics 2.1 Profiles/Screenings – 2.5.3 PCR Profiles – Pig 59 • Bacteriology • Blood, plasma, serum samples Companion Animals 29 • Mycology • Microbiology 2.1.1 Clinical Chemical 2.5.4 Faecal Profiles – Pig 60 • Virology • Urine samples Profiles 29 2.6 Profiles – Hygiene 60 • Parasitology • Histology und immunohistology 2.1.2 Serological Profiles 36 3 Haematology 61 • Maldigestion/Malabsorption • Polymerase chain reaction (PCR) 2.1.3 Travel Profiles and 3.1 Blood Cells 61 • Autovaccines • Immune status Thrombocyte Profiles • etc. Small Animals/ 3.2 Coagulation 63 Tick-borne Diseases 37 3.3 Blood Grouping 66
4 5 2019/20 Contents 2019/20 Contents LABORATORY FOR CLINICAL DIAGNOSTICS
4 Clinical Chemistry 68 15.2 Testing for Specific Parasitic or 20.3.2 Coat Colour/Coat 23.4 Reference ranges farm 4.1 Enzymes 68 Protozoa Infections 244 Structure Cat 326 animals 358 4.2 Substrates 74 15.3 Parasitological Examination – 20.4 Horse 328 23.4.1 Clinical chemistry 358 4.3 Minerals and Trace Minerals 80 Skin 245 20.4.1 Hereditary Diseases 328 23.4.2 Haematological reference 20.4.2 Coat Colour/Coat ranges farm animals 359 5 Urinalysis 86 16 Tests for Indigestion and Structure Horse 335 Diarrhoea 246 20.4.3 Performance Horse 338 24 Conversion Table for Laboratory 6 Allergy 90 16.1 Bacteriological Examination 246 20.5 Cattle 339 Diagnostic Parameters 360 6.1 Allergy Testing 90 16.1.1 Faecal Profiles 246 20.5.1 Hereditary Diseases 339 24.1 Clinical-chemical Parameters 360 6.2 Allergen-specific 16.1.2 Testing for Specific 20.5.2 Breed Characteristics 24.2 Blood Parameters 361 Immunotherapy 95 Indigestion/Diarrhoea Cattle 342 25 Courier Service 362 Pathogens (Bacteria) 250 20.6 Small Ruminants and 7 Immunological Tests/Markers for 16.2 Virological Examination 252 New World Camelids 343 26 Invoicing 362 Inflammation 96 16.2.1 Faecal Profiles – Virology 20.6.1 Hereditary Diseases 343 252 8 Endocrinology/Tumour Markers 103 20.6.2 Breed Characteristics 16.2.2 Testing for Specific Small Ruminants and Indigestion/Diarrhoea New World Camelids 345 9 Function Tests/Calculations 115 Pathogens (viruses) 253 20.7 Pig 345 16.3 Diagnostics of Maldigestion/ 10 Vitamins 127 Malabsorption 254 21 DNA Profile, Breed, Species 346 11 Drug Level 130 16.4 Determination of an Inflammatory 21.1 Identity and Parentage 346 Exudative Process 255 21.2 Breed and Species 347 12 Intoxication 132 17 Autovaccine/Herd-specific 22 Hygiene Examinations 349 13 Infectious Diseases: Pathogenic Vaccine 257 22.1 Profiles Hygiene 349 Agents and Antibody Detection 133 22.2 Single Tests 349 13.1 Viruses 133 18 Pathology 259 18.1 Histopathology 259 13.2 Bacteria 176 23 Reference Ranges 352 18.2 Immunohistology 259 13.3 Fungi 212 23.1 Dog, cat, horse 352 18.3 Cytology 260 13.4 Parasites 215 23.1.1 Clinical chemistry 352 18.4 Lymphocyte Clonality (PARR) 260 23.1.2 Haematological 14 Bacteriology/Mycology 234 18.5 BRAF Mutation 261 reference ranges 14.1 Smears/Puncture 18.6 Differentiation Exsudate/ dog, cat, horse 353 Transudate 261 Fluids/Milk/Faeces 234 23.1.3 Hormones dog, cat, 14.2 Skin/Hair/Feathers 237 19 Sex Determination in Birds 262 horse 354 14.3 Bacteriological Examination 23.2 Reference ranges rabbit, Horse 238 20 Hereditary Diseases/Phenotype/ guinea pig and ferret 355 14.4 Testing for Specific Infectious Breed Characteristics 263 23.2.1 Clinical chemistry 355 Agents 239 20.1 Heredities 263 23.2.2 Haematological 14.5 Sensitivity Testing 241 20.2 Dog 263 reference ranges rabbit, 14.6 Additional sensitivity testing 242 20.2.1 Hereditary Diseases 263 guinea pig, ferret 356 20.2.2 Coat Colour/Coat 23.3 Reference ranges birds 357 15 Parasitology 243 Structure Dog 312 23.3.1 Clinical chemistry 357 15.1 Parasitological Examination – 20.3 Cat 319 23.3.2 Haematological Faeces 243 20.3.1 Hereditary Diseases 319 reference ranges birds 357
6 7 2019/20 Our contacts 2019/20 Our contacts LABORATORY FOR CLINICAL DIAGNOSTICS Our contacts Laboratories Offices
LABOKLIN Germany Tel.: +49 (0) 971 7 20 20 LABOKLIN Belgium Tel.: +32 13 48 05 05 Steubenstraße 4 Fax: +49 (0) 971 6 85 46 E-Mail: [email protected] 97688 Bad Kissingen E-Mail: [email protected] LABOKLIN Croatia Tel.: +385 (0) 91 11 22 121 LABOKLIN Austria Tel.: +43 (0) 732 7172420 E-Mail: [email protected] Paul-Hahn-Straße 3 / BT-D / 1. Stock Fax: +43 (0) 732 717322 4020 Linz E-Mail: [email protected] LABOKLIN Czech Republic Tel.: +42 (0) 730 105 024 E-Mail: [email protected] LABOKLIN Nederlands Tel: +31 (0) 85 4890580 Verlengde Klinkertstraat 6 Fax: +49 (0) 971 68546 LABOKLIN Denmark Tel.: +45 4352 1228 6433 PL Hoensbroek E-Mail: [email protected] E-Mail: [email protected]
LABOKLIN Poland Tel.: +48 (0) 22 691 93 10 LABOKLIN Estonia Tel.: +372 5696 4488 ul. Powstańców Śląskich 101 Fax: +48 (0) 22 691 92 92 E-Mail: [email protected] 01-495 Warszawa E-Mail: [email protected] LABOKLIN Finland Tel: +358 (0) 40 / 7104017 LABOKLIN Slovakia Tel.: +421(0) 948 783 888 Tel: +358 (0) 44 / 0675353 Líščie údolie 57 E-Mail: [email protected] E-Mail: [email protected] 84231 Bratislava LABOKLIN France Tel: +33 (0) 9 67 32 85 80 LABOKLIN Spain Tel.: +34 644 030 557 Fax: +33 (0) 3 88 09 43 82 Polígono Industrial de Alcobendas Tel.: +34 91 198 67 71 E-Mail: [email protected] Avenida de la Industria 4, edificio 3 E-Mail: [email protected] Planta 1ª Oficina A LABOKLIN Iceland E-Mail: [email protected] 28108 Alcobendas (Madrid) LABOKLIN Ireland Tel: +353 19 02 68 06 LABOKLIN Switzerland Tel.: +41 (0) 61 319 60 60 E-Mail: [email protected] P.O. Box Fax: +41 (0) 61 319 60 65 4002 Basel (sample shipment) E-Mail: [email protected] LABOKLIN Italy Tel: +39 (0) 392 033 45 86 Max Kämpf-Platz 1, Erlenmatt, Fax: +39 (0) 051 082 19 75 4058 Basel E-Mail: [email protected]
LABOKLIN United Kingdom Tel.: +44 024 7632 3275 LABOKLIN Latvia E-Mail: [email protected] Batt Laboratories Ltd, E-Mail: [email protected] The Venture Centre University LABOKLIN Lithuania Tel.: +370 6122 2020 of Warwick Science Park E-Mail: [email protected] Sir William Lyons Road, Coventry CV4 7EZ 8 9 2019/20 Our contacts 2019/20 Abbreviations/additional information LABORATORY FOR CLINICAL DIAGNOSTICS
LABOKLIN Norway Tel.: +47 9946 2020 E-Mail: [email protected] Abbreviations Sample material LABOKLIN Portugal E-Mail: [email protected] You will find these abbreviations in part also on our submission forms. The materials required for the individual tests are also indicated on the submission forms, however, for LABOKLIN Russia E-Mail: [email protected] lack of space, not always completely.
LABOKLIN Slovenia E-Mail: [email protected] , Information connected with FACS fluorescence-activated cell comma: you can choose sorting LABOKLIN Sweden Tel.: +46 (0) 723 73 2020 which sample you want to FAVN fluorescent antibody virus E-Mail: [email protected] submit neutralisation + Information connected with FLP fragment length “+”: you must submit all of polymorphism these sample materials FTIR Fourier-transform infrared spectrometry A swab without medium HAH haemagglutination inhibition BAL bronchoalveolar lavage assay CB citrate blood HPLC high performance liquid CFS cerebrospinal fluid chromatography CP citrate plasma ICA immunochromatography EB EDTA blood assay EP EDTA plasma ICPMS inductively coupled plasma FA faeces mass spectrometry IFAT indirect fluorescent antibody H urine technique HB heparin blood ISE ion-selective electrodes HP heparin plasma LCMS liquid chromatography-mass NaFB sodium fluoride blood spectrometry S serum MALDI-TOF matrix-assisted laser TBS tracheobronchial secretion desorption/ionisation – time of TM swab with medium flight mass spectrometry MAT microscopic agglutination test Test methods PARR polymerase chain reaction for antigen receptor rearrange- AAS atom absorption spectrometry ments cELISA competitive ELISA PCR polymerase chain reaction CLIA chemiluminescence assay RIA radioimmunoassay CFT complement fixation test SAF sodium acetate formaldehyde EIA corresponds to ELISA STRs short tandem repeats ELISA enzyme linked immuno www.laboklin.com • www.labogen.com sorbent assay VNT virus neutralisation test
10 11 2019/20 Abbreviations/additional information concerning the test descriptions 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS
Other abbreviations New World camelids llama, alpaca; 1 Pre-analytics AB antibodies Being polygastric, new world bdw body weight camelids may appear under * partner laboratory 1.1 Blood, plasma, serum samples the heading „ruminants“. The first step in the process of examining a sample is the pre-analysis. Pre-analysis Farm includes all steps from patient preparation, specimen collection and transport of the Numbers you may find in the test animals Ruminants and pigs sample to the lab to the preparation of the sample for analysis. descriptions (1) Specifications apply to testing 1.1.1 Preparation of the patient Further notes with method (1) Before taking a blood sample, the patient should normally fast for 10 - 12 hours, (2) Specifications apply to testing The obligation to notify the authorities upon provided the physiology of the species concerned permits this. Otherwise, faulty results with method (2) suspicion of a disease applies to Germany. are to be expected, especially for cholesterol, glucose and TLI. In addition, parameters (3) Specifications apply to testing The obligation to notify the authorities upon such as α-amylase, ALT, AST, bilirubin, total protein, triglycerides, serum bile acids, with method (3) diagnosis of a disease applies to Germany. leukocytes and calcium can be affected. Fasted blood samples are not unproblematic in horses and only indicated for special > greater than tests (e.g. insulin and glucose for the diagnosis of the Equine Metabolic Syndrome). Duration The information applies from < less than It is advisable to inform the owner about the influence of physical activity or stress on the date of arrival of the the results of a blood examination. Particularly muscular enzymes such as CK, LDH samples at Laboklin. and AST can show increased levels in the serum after physical exertion. Additionally, ”Days” means ”working days”. glucose and lactate can also show elevated serum levels. Due to delays in transport, test duration may take longer Before doing any allergy tests, including feedstuff tests, any administration of for tests which are carried out corticosteroids should be stopped. To do so, the following withdrawal times are by a partner laboratory. recommended: The specified test durations - local/topical corticosteroids: 2 - 4 weeks are supplied without liability. - oral corticosteroids (e.g. Prednisolone): up to 8 weeks - depot cortisone preparations (e.g. Voren®): up to 3 months If these times cannot be observed, false negative results are possible. In the case of a Species positive result, the reaction class must be assessed taking into account the previous Large administration of cortisone. Please note that other itch-suppressing medication may animals Horses and farm animals also have a negative impact on the allergy test. Our allergy team will be pleased to advise you. Small mammals Rabbit, guinea pig, rat, 1.1.2 Which sample? mouse, hamster, ferret and other small mammals which Details on the recommended material (blood, serum, plasma) for the requested test are being kept as pets can be taken from our test descriptions or the submission form. For labelling the In individual cases, tests sample, it is also necessary to indicate the sample type (see Chapter 1.9 and 1.10). for small mammals may be also applicable for small wild Whole blood samples mammals (e.g. hedgehogs). EDTA blood (EB) Small - For doing a blood count, EDTA blood is the most suitable material in mammals animals Dog and cat (however, for birds and reptiles it is heparin blood, see below). - For the serological examination of the blood type, EDTA whole blood is needed as well.
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- As the cells in the sample are not stable, EDTA samples for haematological tests Serum should not be older than 48 hours. - Samples are drawn into tubes without anticoagulants. - For most PCR analyses and genetic tests, EDTA blood is required. - Allow to stand for 30 – 60 min. - To determine certain parameters such as ACTH or pro-BNP, only EDTA plasma which - Centrifuge for 10 min at 2000 g. was promptly centrifuged and cooled can be used to obtain reliable results. - Remove the supernatant by pipette and transfer it into an uncoated test tube, then label the test tube. Heparin blood (HB) - For correctly determining individual parameters like insulin or fructosamine, only - To collect heparin samples, lithium heparin (LiHep) tubes are available. serum should be used. - For doing a blood count for reptiles and birds, lithium heparin blood should generally - Sending non-centrifuged samples should only be done exceptionally (e.g. in case of be used. a very low sample quantity), as the transport might result in cell damage and thus lead - Since the amount of blood is often very low in small mammals, lithium heparin tubes to a haemolytic serum. are particularly suitable, as they cannot only be used for doing a blood count but also for determining a wide range of blood-chemical parameters. - For the PCR, lithium heparin whole blood should only be used under exceptional 1.1.3 Factors interfering with analysis circumstances, as lithium heparin can inhibit the PCR and might thus lead to false negative results. Haemolysis Haemolysis is caused by leakage of intracellular components of the erythrocytes such Citrate blood (CB) as phosphate, iron, potassium and especially haemoglobin due to a damage of the cell - To determine the coagulation parameters, only the appropriate citrate tubes should be membrane. Haemoglobin causes a red colouration of serum/plasma which primarily used. For getting a correct evaluation, their shelf life may not be exceeded. It is also interferes with the photometric testing done in clinical chemistry. necessary to have an exact mix ratio of 1:10 (1 part citrate + 9 parts blood). - Most coagulation parameters can be analysed using citrate whole blood. Lipaemia - For correctly performing platelet function tests, citrate whole blood is required. Lipaemia refers to the milky/turbid discolouration of serum/plasma due to triglycerides. It is mostly caused by diet- or stress-related factors. Lipaemia can also occur as a result Sodium fluoride blood (NaFB) of endocrinological diseases like Cushing’s syndrome or hypothyroidism. Sodium fluoride inhibits enzyme activities which lead to a reduction of some parameters. It should be used for the correct determination of glucose and lactate. Lipaemic samples often complicate the measurement of certain clinical parameters, e. g. bilirubin. Plasma Icterus - Samples are drawn into tubes with anticoagulants (heparin, EDTA, citrate). Icterus is a yellowish discolouration of serum/plasma. Excess amounts of bilirubin, - Can be centrifuged immediately after collection (10 min, 2000 g). which is the reason for the yellow colouring, are normally caused by a medical condition - Remove the supernatant by pipette and transfer it into an uncoated test tube, then and cannot be influenced. indicate the sample materials on the test tube or use the appropriate bar code label. - Please note: The additives limit the number of analyses! The yellow colouration is physiological in horses. - Heparin plasma (HP) is needed for many clinical-chemical examinations. HP cannot be used for agglutination tests. - The collection of EDTA plasma (EP) for clinical-chemical and/or serological parameters should only take place in exceptional cases, as EDTA disturbs through various mechanisms the measurement of individual parameters such as calcium, magnesium and AP. Likewise, potassium cannot be determined when using EDTA plasma, since EDTA is added as K-EDTA. - Some coagulation parameters can only be analysed using citrate plasma (CP). Performing platelet function tests using centrifuged citrate plasma is not possible.
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Interfering factor Parameter Level 1.1.4 Specific features Haemolysis LDH, CK, AST, bilirubin, AP, creatinine, PO4, K, Mg, Blood counts Fe, fructosamine - EDTA or lithium heparin blood Haemolysis Ca, glucose - When collecting the sample, discard the first 0.5 ml of blood, if possible, as they contain an increased amount of coagulation factors, or first obtain a serum sample. Lipaemia ALT, AST, AP, bilirubin, glucose, Ca, PO4, - Let the blood run down slowly on the side of the sample tube. total protein, lipids, haemoglobin - Pay attention to the fill volume! Preferably fill up to the mark, since an insufficient Lipaemia albumin, amylase, Na, CI, K volume can result in changes in cell morphology. Do not overfill the tube in any case, as the sample might clot. Icterus AP, total protein, Cl, PO4 - After drawing the sample, tilt the test tube carefully several times. - Do not store blood smears in the refrigerator and not close to formalin. Icterus triglycerides, creatinine, Mg - Pack the samples frost-proof in winter; possibly cool them in summer. - Reliable results can only be obtained from samples not older than 48 hours.
Clinical chemistry of serum or heparin plasma Medicine Parameter Level Prompt centrifugation of the samples lead to better test results, as it reduces the risk of Penicillin G K haemolysis caused by transportation. However, serum should be allowed to stand for a minimum of 30 min to ensure a complete clotting of the sample. Tetracyclines PO4 Serum samples can also be shipped frozen; they will then reach the laboratory cooled. Repeated freezing/thawing, though, should absolutely be avoided. Tetracyclines K Determination of glucose and lactate Salicylates CK, AP, glucose, Na, total protein - Requires sodium fluoride blood or sodium oxalate blood Salicylates K, Ca - Fill the test tube up to the mark at the maximum. Corticosteroides CK, AP, glucose, Na, total protein Coagulation parameters - Determination is carried out using sodium citrate plasma which is obtained from Corticosteroides K, Ca citrate blood with the mix ratio being 10:1 (9 parts blood + 1 part sodium citrate). Centrifugation should take place at the practice. Phenylbutazone Ca, Na - If commercial citrate-treated tubes are used, the expiry date needs to be checked before collecting the sample. Expired tubes may no longer be used, as skewed results Barbiturates CK are to be expected. When drawing the sample, special attention must be paid to the Halothane CK, PO4 exact fill level (marking on the tube). anaesthesia - If no commercial tubes are available, sodium citrate 3.13% can be drawn into a syringe. - No heparinised needles or catheters may be used. Glucose infusion glucose Glucose infusion PO4 1.2 Microbiology
- It is important to collect samples as sterile as possible to avoid contamination with physiological and/or ubiquitous flora. - Smears for bacteriology (aerobic and anaerobic microbes) and for mycology should be sent with a transport medium (“swab with medium” = TM) to protect the microbes during shipment.
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- Smears for examinations using PCR should be sent without a transport medium Detailed explanations: (“smear without medium” = A, “swab without medium”, “dry swab”) or with a special As a sample, a representative piece of tissue free of preparation artefacts (e.g. medium. disruption, squashing, electrocoagulation) should be taken. The diameter of the sample - Urine samples should be sent in a sterile tube using Uricult or using a swab with should not be less than 0.5 cm. An exception to this are samples which, for technical transport medium. reasons, cannot be obtained otherwise (such as endoscopically taken stomach - Hairs and/or skin scrapings for the diagnosis of dermatophytes are best sent in a biopsies). Furthermore, it should be borne in mind that samples which are too small paper bag or in aluminium foil. only provide little information, whereas samples that are too large cannot be fixed - For sending faeces/excrement, special transport tubes should be used, no bags or properly. Pieces of tissue with an edge length of 1 cm are recommended. However, gloves tied with a knot; for birds, use cloacal swabs with transport medium. this might vary depending on the lesion to be examined, the sampling site and the - Blood culture flasks can be ordered with prior written notice (subject to a objective. charge). Small lesions should be placed centrally so they are not overlooked and thus truncated during preparation. If in doubt, several samples should be collected.
1.3 Hygiene 1.4.1 Skin punches As skin samples, punch biopsies of all dermal layers with a diameter ≥ 0.6 cm are to The contact plates required for examining the surface disinfection must be ordered be submitted. Primary lesions from several locations should be selected. The biopsied from the laboratory in good time, as they can only be stored for a short period of time. area should not be pre-treated by scraping or shaving. The anamnesis should contain all The surfaces to be sampled have to be cleaned and disinfected and must be well dried relevant data which might be important for the diagnosis. It is recommended to use our before sampling. The contact plates, which need to be labelled on the bottom, must be submission form Pathology, which especially focuses on skin and tumour diagnostics, returned cooled to the laboratory within 24 hours together with the filled-in submission but also leaves room for any other type of anamnesis. form. The bioindicators necessary for evaluating the proper functioning of the sterilisers 1.4.2 Cytology should be requested from the laboratory as needed and can be applied right in the Samples can primarily be taken as wipe test, scraping or puncture (with or without device together with the regular sterilisation material. After testing, these bioindicators aspiration). The most common technique is the fine needle aspiration, using a thin have to be sent directly to the laboratory together with the filled-in submission form. hollow needle (G 22 – G 27) attached to a syringe. A vacuum is created and, if possible, For testing the disinfection of endoscopes, two contact plates and two rinse samples the tissue should be punctured several times in different directions. Before detaching are necessary for each endoscope. Before testing, the material must be requested the needle, the vacuum must be released to avoid the material receding into the from the laboratory in due time. A detailed description of all the processes of taking the syringe. The material obtained is then pressed out of the needle onto the side of a glass sample will be provided. It is required to return the collected samples to the laboratory slide. A second slide is placed flat at a right angle on top of the first one and is then within 24 hours. carefully pulled away across the slide. If the sample is more liquid, a steeper angle (45°) – like in a blood smear – should be applied. For the cytological examination of aspirates, excretions or secretions, the fluids 1.4 Histology and immunohistochemistry obtained are centrifuged at 2500 – 3000 rpm for three to five minutes. The supernatant is decanted and the sediment is carefully spread like a blood smear and shipped air- When submitting tissue samples for histopathological and immunohistochemical exa- dried. If the aspirates are sent directly, EDTA tubes should be used as test vessels. minations, the following points must be observed: For bronchial, conjunctival and vaginal cytology, the swab obtained (cytobrush) should - artefact-free extraction of a typical lesion sufficient in size (diameter > 0.5 cm) be rolled onto a glass slide, not smeared. - immediate fixation (4% neutral buffered formaldehyde 10% formalin) All smears should generally be sent in air-dried, but unfixed and unstained. The most - preparation of an anamnesis including objective and clinical picture important point is to create a thin smear consisting of only one layer (monolayer). The - shipment in a suitable container (available from us free≙ of charge) most common reason for getting a limited quality up to not being able to assess at all - Immunohistochemistry can always be done after histopathology with the material are smears that are too thick. supplied.
18 19 2019/20 Pre-analytics 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS 1.5 Polymerase chain reaction (PCR) swabs (without transport medium) should be supplied. To create DNA profiles and parentage reports in dogs and cats, we recommend to always send in a blood sample. PCR is a very sensitive and specific method for the direct detection of infectious For all genetic testing in horses, it is sufficient to supply about 20 hair roots from mane agents. Via PCR, gene sequences characteristic for the respective pathogen are or tail for DNA isolation. reproduced and detected – if necessary, even of germs which are no longer viable. EDTA blood is the most suitable sample material. It is absolutely essential to use EDTA The sample material that must be supplied for the PCR highly depends on the as anticoagulant. Lithium heparin or citrate are unsuitable as anticoagulants, as they pathogen to be detected and the present symptoms or the objective. Depending on may inhibit the subsequent PCR. In very rare cases, haemolysis induced by transport how the pathogen has spread in the body and its excretion, different sample materials or extreme stress during sample collection might lead to the situation that no result can are suitable. be obtained. However, the percentage of blood samples which cannot be evaluated is extremely low, being < 1%. At this stage of infection, pathogens causing viraemia, parasitaemia or bacteraemia can be detected directly in an EDTA blood sample (EB). Lithium heparin is less suitable as Buccal smears, often incorrectly called saliva samples, are very suitable sample materi- an anticoagulant, as it can inhibit the PCR. For blood samples or other liquid samples, als for genetic testing in dogs and cats, as long as the sampling procedure is performed an amount of at least 0.2 ml is required. correctly observing the following rules: 1. The animal should not have eaten anything for about 1 hour prior to the sample collec- In contrast to cultural bacteriological/mycological examinations, for PCR tests it is tion. It should be ensured that puppies and kittens have not been nursed for a minimum recommended to use sterile swabs without transport medium (“smear without of 2 hours, as otherwise maternal cells might skew the results. medium” = A, “swab without medium”, “dry swab”). If the concentration of the 2. When taking the sample, it should be scrubbed strongly at the inside of the cheek to pathogen is low, smears in a medium can lead to false negative results. For collecting make sure that enough cells of the oral mucosa and thus genetic material is attached to the sample, the swabs can be moistened with physiological saline solution. For PCR the swab. Genetic testing can only be conducted if enough genetic material adheres to tests, so-called cytobrushes (brush swabs) are also suitable, which can be shipped in the swab. Generally, saliva alone is not sufficient. However, there should not be any blood an uncoated sterile tube. on the swabs! 3. In order to prevent the growth of bacteria and mould, the swabs should be dried for For the detection of pathogens in faeces, a sample of approximately the size of a about 2 – 4 hours after collecting the sample. This is done best by keeping the test tubes hazelnut is needed. For some agents (e.g. coronavirus, Tritrichomonas foetus) we a little open for a while. recommend collecting faeces samples for 3 days, since these pathogens are excreted As there is considerably less cell material available from mucosal swabs compared to intermittently in the faeces. blood samples, it is not always possible to isolate enough DNA from buccal smears for Further sample materials, e.g. skin biopsies, organ material, urine, synovial fluid, liquor, a genetic test. This applies to about 5% of the submitted buccal swabs. We recommend bone marrow aspirates and lymph node aspirates, for PCR tests are best sent in sterile, sending two buccal samples per animal, so there is more material available for testing. uncoated test vessels. Fixation solutions such as formalin or the like can lead to DNA degradation, PCR inhibition and thus to false negative results. For horses, hair roots can be used to perform genetic examinations. To do so, about 20 Samples do not normally need to be sent cooled. Until it is dispatched, the sample pulled mane or tail hairs are needed. If samples are taken from various animals, hands material can be stored in the refrigerator at 2 – 8 °C. Repeated freezing/thawing should must be cleaned thoroughly after each sampling – even a single hair of a different animal absolutely be avoided. can skew the result. Hairs can, for instance, be shipped in little plastic bags or in envelopes. It is, however, Please note: Creating an antibiogram is not possible after doing a PCR test. absolutely necessary to make sure that the hairs are put in a closed envelope, separate from the submission form, when sent in.
There should not be any blood samples sent in for cattle from multiple births because of 1.6 Genetic testing a possible blood chimerism, but if the test allows it, hair roots, sperm or tissue samples can be used. One exception to this is the freemartin test, for which a blood sample is As sample material for the molecular genetic detection of hereditary diseases, for paren- mandatory. tage analysis as well as for the genetic determination of coat colours and blood types, If you wish to supply sample materials different from those listed above for performing EDTA whole blood samples (approx. 1 ml) are suitable. Alternatively, in dogs and genetic tests, please contact us before sending the samples. cats, buccal swabs, so-called cheek swabs, can be used. For each animal, 2 buccal
20 21 2019/20 Pre-analytics 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS 1.7 Immune status (5) Citrate tube# (11) Shipping container for CB = Citrate blood: It can be swabs with/without medium EDTA blood samples must not be older than 48 hours! shipped in this tube (+ No. 8). CP = Citrate plasma: The sample should be centrifuged and the supernatant needs to be transferred 1.8 Sample material/Shipping material into a neutral tube (e.g. Eppendorf tube). It must then be marked accordingly as CP. (12) Urine tube Note regarding the sample materials listed in the test descriptions from Chapter 2 onwards: (suitable shipping container If the abbreviations are separated by a comma, you can choose the material which is (6) Salivette® see No. 8) easiest for you to collect from the given list. When collecting sample material for the for collecting saliva samples detection of a pathogen using PCR, you should preferably collect that material from the listed alternatives which is likely to have the highest concentration of pathogens. If the specifications are connected by one or more “+”, both or all the materials joined with “+” need to be provided for determining all the parameters of the selected testing (13) Container for histology block. (7) Blood smear (formalin tube with The materials required for the individual tests are also indicated on the submission Blood smears should always shipping container) forms, however, for lack of space, not always completely. be sent in air-dried, unfixed and unstained. For transportation, the Sample and shipping containers that are available for the collection and transport of depicted transport covers (shipping (14) Faeces tube the samples include: containers) are suitable. Before with shipping container transport, store at room temperature (25 °C) (may not be cooled).
(1) EDTA tube# (3) NaFB = Sodium fluoride blood (8) Shipping containers for EB = EDTA blood: It can be With NaFB samples, too, please pay blood tubes or urine tubes (15) Blood culture flask shipped in this tube (+ No. 8). attention to the labelling. EP = EDTA plasma: EDTA blood has to be centrifuged and the supernatant needs to be trans- ferred into a neutral tube (e.g. Eppendorf (4) S = Serum# (9) TM = Swab with tube). It must then be marked accordingly To collect serum, the coagulated transport medium # On special request, small test tubes as EP or labelled with the appropriate bar blood should be centrifuged at (orange: thin swab, (see each of the tubes shown on the left) code. 2000 g 30 minutes after being black: thick swab) are provided for collecting small amounts collected. The supernatant should of EDTA blood, EDTA plasma, heparin (2) Heparin tube# then be transferred into a neutral blood, heparin plasma, citrate blood, HB = Heparin blood: It can be tube or another serum tube (remove citrate plasma and serum, e.g. from small shipped in this tube (+ No. 8). beads before!) and marked accordingly mammals. If required, please order these HP = Heparin plasma: Heparin as serum or labelled with the appropriate small test tubes by e-mail or telephone blood has to be centrifuged and bar code. only. the supernatant needs to be transferred (10) A = Swab At your express request (information by into a neutral tube (e.g. Eppendorf tube). without transport medium, telephone or e-mail), we can also send It must then be marked accordingly as HP. (dry swab) you vacuum tubes for the examination of blood samples at Laboklin.
22 23 | 2019/20 Pre-analytics 2019/20 Pre-analytics |} LABORATORY FOR CLINICAL DIAGNOSTICS 1.9 Labelling Labeling of samples and shipping materials - The names of the animal and the owner should be clearly marked on the submission form and the sample. Alternatively, bar code labels can be used to unmistakably iden- Step 1 Step 2 1< GRG fq| G|