FAQs from the consult archives Emmeline Tan, DVM, DVSc, Dipl. ACVP (Clinical Pathology) October 29, 2015
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1 FAQs from the consult archives Emmeline Tan, DVM, DVSc, Dipl. ACVP (Clinical Pathology) October 29, 2015
2 Today: Some tips to help maximize value of your cytology submissions Only if we have time: Diagnostic testing for infectious diseases: It s alphabet soup and I m not sure which test to pick
3 Cytology interpretations we all hate! Hypocellular sample Insufficient cellularity Sample unsuitable for cytologic evaluation Inconclusive = Non-diagnostic sample Not enough info to render interpretation Increase costs of testing (wasted time, $)
4 Tips for cytology submissions: 1. Know the limitations of cytology No tissue architecture Canine mammary neoplasms Keratin-producing, follicular/adnexal tumors Feline lymphoma, canine indolent nodular lymphomas Most oral/gingival lesions Some lesions just don t exfoliate well & are not amenable to cytologic diagnosis! Some mesenchymal neoplasms Very small tumors Manage your (and your client s) expectations!
5 Tips for cytology submissions: 2. Provide a good history The Influence of Clinical Information in the Histopathologic Diagnosis of Melanocytic Skin Neoplasms Ferrara G, Argenyi Z, Argenziano G, Cerio R, Cerroni L, et al. (2009) PLoS ONE 4(4): e5375. doi: /journal.pone
6 Tips for cytology submissions: 2. Provide a good history Clinical information IMPROVES test reading accuracy (the bias argument is bogus) No Hx: Clinical pathologists are MORE likely to hedge, LESS likely to provide definitive diagnosis (clinical impact of diagnosing malignancy in bronchial brush specimens, for ex.) Some instances: definitive Dx = clinicallyaided
7 Tips for cytology submissions: 2. Provide a good history Acellular slides Soft, subcutaneous and mobile mass over left thorax. Skin and hair over top are normal. No history provided Compatible with lipoma Non-diagnostic sample
8 Tips for cytology submissions: 3. Spread your samples gently! Cytological assessment requires a monolayer of intact cells. Poorly spread samples are too thick to discern cytological detail. Samples just deposited onto slide without spreading
9 Tips for cytology submissions : 3. Spread your samples gently! Examples of well-spread slides (L-R): BM aspirate; LN aspirate; aspirate of hairless skin mass on pinna of dog
10 Tips for cytology submissions : 3. Spread your samples gently! Over-exuberant squishing may rupture cells Lymphoid tissue Round cell tumors Especially LN aspirates: It s a matter of balance, and lots of practice!
11 Tips for cytology submissions: 4. Check that slides are cellular Unless you suspect lipoma Diff Quick slide should be submitted along with your unstained slide(s) Why do clinical pathologists prefer Wright or Wright- Giemsa?
12 Tips for cytology submissions: 5. Keep formalin far, far away Photos courtesy Dr. Dorothee Bienzle, Dept. of Pathobiology, University of Guelph
13 Tips for cytology submissions: 6. Use pencil to label slides Pen and Marker ink (most indelible ones, too) wash away during staining
14 Maximizing cytology submissions: 7. Don t be afraid to ask! If you are unsure it is worthwhile to do FNA or have any other questions about submitting a sample feel free to ask clinical pathologist!
15 Any questions?
16 Infectious disease testing: Swimming in alphabet soup! ELISA IFA PCR MAT WB Leptospira Yes (Ab) x Yes Yes (Ab, Gold standard) FeLV Yes (Ag) Yes (Ag) Yes x x FIV Yes (Ab) Yes (Ab) Yes x Yes (Gold standard) Hemotropic Mycoplasma x x Yes x x Toxoplasma Yes (Ab) Yes (Ab) Yes Yes (Ab)** x x
17 Infectious disease testing: Really only 2 approaches Detecting infectious agent: Proteins produced by agent IFA, ELISA DNA of the agent PCR The whole agent culture or virus isolation Detecting host immune response against agent: Antibody detection IFA, ELISA, agglutination, Western blot
18 Infectious disease testing: ELISA Enzyme-conjugated antibody that recognizes Ag or Ab Substrate provided, enzyme catalyzes reaction coloured product is detected
19 Infectious disease testing: IFA Fluorofore-conjugated antibody that recognizes Ag or Ab Exposure to specific wavelength of light fluorofore excitation and re-emission of light Detector system picks up this signal
20 Infectious disease testing: WB WESTERN BLOT (IMMUNOBLOT)
21 Infectious disease testing: MAT AGGLUTINATION TESTS Mostly for detection of antibodies
22 Infectious disease testing: PCR 15PCR.htm Amplification of a target DNA region Primer: starting point, determines target DNA region amplified Also need nucleoties, some heat, and stable DNA polymerase
23 So, in conclusion Methods to detect infectious agent may be affected by prior treatment (which may reduce the actual # of bugs that can be detected or their viability) Methods to detect antibody response may be affected by prior vaccination or alterations in immune response Except not Lyme QuantC6 (based on surface protein ONLY produced by Borrelia when it is inside the mammalian host; not when organism is within tick or used to make the Vx) Titres may be useful to differentiate prior vaccination from true infection
24 Example: Leptospirosis Methodology Pros Cons MAT Considered gold standard Vx can cause positive but have idea if probable infection or prior Vx (actual titre provided) Long TAT (labor intensive) Have to keep live leptospires Expensive Acute/convalescent titres req d ELISA Quick TAT, inexpensive Pos/neg only no titre (no differentiation recent Vx vs. true infection) PCR Quick TAT compared to MAT, inexpensive No interference with prior Vx Need paired urine/blood samples False negative with prior antibiotic treatment, aged samples (DNA breakdown)
25 Rule in/out leptospirosis: Which test(s) would you pick? 10 y.o. FS mini schnauzer Acute onset vomiting (3 dd), lethargy, inappetance, ~10% dehydration, mild fever edvm did in-house CBC/chem, started IVF, ABx on intake transferred to rdvm the next day CBC: Neutrophilic left shift with toxicity, monocytosis Vx (DHPP+L) 4 months ago
26 Any questions?
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