SEROLOGYALLERGY TESTINGS OTHER OPTION

OVERVIEW

WHAT IS AN ALLERGY? SYMPTOMS SUPPRESSION VS HYPOSENSITIZATION WHEN TO ORDER TYPES OF TESTING HOT TOPICS IN ALLERGIES TODAY Q&A

WHAT IS AN ALLERGY?

SYMPTOMS:

SKIN ISSUES

RESPIRATORY ISSUES

DIGESTIVE PROBLEMS

OCULAR SYMPTOMS

SKIN ISSUES

ITCHING

REDNESS

HOTSPOTS

HAIR PULLING

RESPIRATORY ISSUES:

COUGHING

SNEEZING

WHEEZING

REVERSE COUGH

DIGESTIVE ISSUES:

VOMITING

DIARRHEA

SYMPTOMS OVERVIEW:

SUPPRESSION VS HYPOSENSITIZATION SUPPRESSION:

STEROIDS CYCLOSPORINE ANTIHISTAMINES

HYPOSENSITIZATION: SMALL, CONTROLLED AMOUNTS OF

OFFENDING ALLERGENS GIVEN SUBCUTANEOUSLY IN INCREASING DOSES AND FREQUENCY; PLANT DERIVED.

SUPPRESSION SIDE EFFECTS

STEROIDS DOGS:

WEIGHT GAIN, HYPERACTIVITY, PANTING, DIARRHEA & DEPRESSION

LIVER ADRENAL GLANDS PANCREAS DIABETES

CATS: REQUIRE HIGHER DOSES TO ACHIEVE RESPONSE

=COST LESS LIKELY TO DEVELOP ADVERSE SIDE EFFECTS

SUPPRESSION SIDE EFFECTS ANTIHISTAMINES

ONLY ABOUT 1/3 OF PET PARENTS REPORT SUCCESS

TRY SEVERAL DIFFERENT TYPES BEFORE GIVING UP

COMMON SIDE EFFECTS CAN RANGE FROM DROWSINESS TO SEIZURES TO NAUSEA.

SUPPRESSION SIDE EFFECTS CYCLOSPORINE

FEW KNOWN SIDE EFFECTS TO DATE, LONG TERM RISKS STILL UNCERTAIN

COMMON SIDE EFFECTS INCLUDE: VOMITING, NAUSEA, SOFT STOOLS

LESS COMMON HEPATITIS, BONE MARROW SUPPRESSION, SEIZURES HAVE BEEN REPORTED.

STEROID’S LEGACY

1. SKIN

INCREASED SUSCEPTIBILITY TO INFECTION; THINNING OF THE SKIN.

2. ADRENAL

CUSHING’S SYNDROME

3. GASTROINTESTINAL

VOMITING, DIARRHEA, ULCERS

4. KIDNEY

RENAL COMPROMISE, ELECTROLYTE IMBALANCE, INCREASED THURST/URINATION

5. MUSCLE

WASTING AND WEANESS, PENDULOUS ABDOMEN

6. PANCREAS

TYPE 2 DIABETES

PRELIMINARY STEPS

MEDICAL HISTORY AGE OF SYMPTOMS ONSET

ROUTE OF POSSIBLE EXPOSURE

RELATIONSHIP FOOD/ENVIRONMENTAL EXPOSURE

RESPONSE TO ADMINISTERED TREATMENT

(HANDOUT)

PRELIMINARY STEPS CONTINUED… PHYSICAL EXAMINATION

TRACK CLINICAL SIGNS (SEASONAL, PERENNIAL)

ADDRESS ANY SECONDARY INFECTIONS DISTINGUISH SIGNS FROM OTHER DISEASE

INITIAL LABORATORY TESTS IF NECESSARY TO RULE OUT SYSTEMIC

CONDITIONS

(HANDOUT)

ALLERGY TESTING

WHEN TO CONSIDER AN ALLERGY WORKUP:

PET PARENT IS COMMITTED TO TREATMENT PROCESS REALISTIC RESULTS GOOD USE OF FUNDS

CONFIRM SUSPECTED DIAGNOSISNOT FIRST LINE OF DEFENSERULE OUT OTHER IMMUNOLOGICAL

ISSUES FIRST

TYPES OF TESTING

INTRADERMAL

SEROLOGICAL

INTRADERMAL TESTING

WEEN OFF SUPPRESSION THERAPY (2-6wks)

TWO+ VISITS TO DERMATOLOGIST ANIMAL SEDATED INJECT SMALL AMOUNT OF ALLERGEN MEASURE RESULTS

http://www.vlsstore.com/Media/PublicationsArticle/Intradermal.pdf

SEROLOGY

SIMPLE BLOOD TEST NO NEED TO WEEN OFF SUPPRESSION

THERAPY CAN BE DONE IN 1 VISIT, IN CLINIC COST-EFFECTIVE OPTION RISK REDUCTION TEST FOR FOOD REACTIONS

TYPES OF SEROLOGY

WESTERN BLOT

RAST

ELISA

MODIFIED ELISA

WESTERN BLOT

PROCESS Proteins are separated by gel electrophoresis. Proteins are transferred to a sheet of special blotting

paper called nitrocellulose. The proteins retain the same pattern of separation they had on the gel.

The blot is incubated with a generic protein, to bind to any remaining sticky places on the nitrocellulose.

An antibody is then added to the solution which is able to bind to its specific protein. The antibody has an enzyme or dye attached.

The location of the antibody is revealed by incubating it with a colorless substrate that the attached enzyme converts to a colored product that can be seen and photographed.

WESTERN BLOT CONTINUED…

WESTERN BLOT CONTINUED… PROS

RELATIVELY CHEAP & FAST METHOD DETERMINE AMOUNT & SIZE OF PROTEINS

CONS QUESTIONS RAISED ABOUT RELATIVE

ABUNDANCE OF PROTEINS WITHIN A TISSUE ELECTROSTATIC MIGRATION VARIATIONS

http://teachline.ls.huji.ac.il/72320/methods-tutorial/western/western-pros.html

www.abcam.com/technical (other troubleshooting suggestions)

RAST (RADIOALLERGOSORBANT ASSAY)

PROCESS USES A RADIOIMMUNOASSAY TO DETECT SPECIFIC

IgE ANTIBODIES. THE SUSPENDED ALLERGEN IS BOUND TO AN

INSOLUBLE MATERIAL AND SERUM IS ADDED. IF SERUM CONTAINS ANTIBODIES TO ALLERGEN

THOSE ANTIBODIES WILL BIND TO THE ALLERGEN. RADIOLABELED ANTI IgE IS ADDED WHERE IT

BINDS AND ALL UNBOUND IgE IS WASHED AWAY. THE AMOUNT OF RADIOACTIVITY IS

PROPORTIONAL TO THE SERUM IgE FOR THE ALLERGEN.

RAST CONTINUED…

RAST CONTINUED…

PROS IMPROVED SENSITIVITY WITHOUT LOSING

SPECIFICITY REPRODUCIBILITY

CONS COST AVAILABILITY OF RADIOIMMUNOASSAY POTENTIAL RISK TO USER DUE TO

MATERIALS

ELISA (ENZYME LINKED IMMUNOASSAY)

PROCESS INVOLVES AT LEAST 1 ANTIBODY WITH

SPECIFICITY FOR A PARTICULAR ANTIGEN. ANTIGEN IS IMMOBILIZED ON A SOLID SUPPORT

(MICROTITER PLATE). DETECTION ANTIBODY IS ADDED, FORMING A

COMPLEX WITH THE ANTIGEN. BETWEEN STEPS PLATE IS WASHED TO REMOVE

ANY PROTEINS/ANTIBODIES NOT SPECIFICALLY BOUND.

ENZYMATIC SUBSTRATE PRODUCES VISIBLE SIGNAL.

ELISA CONTINUED…

ELISA CONTINUED…

PROS NO RADIOACTIVITY NEEDED SAFE INEXPENSIVE

CONS MORE DIFFICULT TO DEVELOP, CONTROL &

STANDARIZE HIGHER PROBABILITY OF FALSE NEGATIVES

MODIFIED ELISA

PROCESS: THE PROBLEM WITH A STANDARD ELISA

ASSAYS IS THAT IN THE WASHING PROCEDURES, THE ALLERGEN BOUND TO THE PLATE WASHES OFF AND CONSEQUENTLY WHEN THE PATIENT’S ANTIBODY IS ADDED THERE IS NO ANTIGEN THERE, THUS A FALSE NEGATIVE RESULTS.

SPECTRUM’S SPOT TEST ADDRESSES THIS BY USING A PATENTED TECHNOLOGY TO COVALENTLY BIND ALLERGEN TO THE PLATE MUCH LIKE Ag IS BOUND TO THE CELLULOSE DISC IN THE RAST.

MODIFIED ELISA CONTINUED…

MODIFIED ELISA CONTINUED… PROS

(SPECTRUM) RELIABILITY OF RAST WITH THE ECONOMY OF SCALE OF THE ELISA

COVALENT BONDING

CONS STILL CONFUSED WITH STANDARD ELISA’S

FALSE NEGATIVE RESULTS RELATIVELY NEW TECHNOLOGY, STILL IN

REVIEW

THINGS TO CONSIDER

BOTH RAST AND ELISA ARE ESENTIALLY USING THE SAME PRINCIPAL IN THAT THEY BOTH RELY ON AN ANTIGEN ANTIBODY REACTION OCCURING WITH AN ANTIGEN BOUND TO A MATRIX; LIQUID IN RAST, SOLID IN ELISA.

SOMETIMES SOURCE1 FINDS ONE OUTCOME AND SOURCE2 ANOTHER. BOTH HOWEVER CAN BE ACCURATE.

HYPOSENSITIZATION

SUCCESS RATES AS HIGH AS 85-90% REDUCTION IN SYMPTOMS FROM

DIETARY CHANGE ALONE =40-50% IMPROVED QUALITY OF LIFE FOR PETS PLANT DERIVED SAFE FOR LONG TERM USE LONG TERM ALLERGY MANAGEMENT

TOOL NOT CURE

HYPOSENSITIZATION BEFORE & AFTER

HOT TOPICS IN ALLERGY

TEWL STUDIES

CORELATION BETWEEN TESTING METHODS

RUSH IMMUNOTHERAPY

Q&ASESSION

SOURCES: SLIDE 3: VIDEO

DR. RUFUS RAJADURAI, M.D.; www.rufusrajadurai.wetpaint.com

SLIDE 10: SUPPRESSION SIDE EFFECTS DR. MIKE RICHARDS, DVM http://www.vetinfo.com/ceffect.html DR. BARBARA FORNEY, VMD http://www.wedgewoodpetrx.com/learning-center/professional-monographs/prednisone-for-veterinary-use.html DR. RON HINES, DVM PhDwww.2ndchance.info/cushings.htm

SLIDE 18: INTRADERMAL TESTING http://www.vlsstore.com/Media/PublicationsArticle/Intradermal.pdf

SLIDES 21-32: SERUM TESTING METHODOLOGIES http://en.wikipedia.org/wiki/ELISA http://en.wikipedia.org/wiki/RAST_test http://en.wikipedia.org/wiki/Western_blot http://teachline.ls.huji.ac.il/72320/methods-tutorial/western/western-pros.html

SOURCES CONTINUED

SLIDE 34: HOT TOPICS IN ALLERGY TEWL (TRANSEPIDERMAL WATER LOSS

http://publik.tuwien.ac.at/files/pub-et_11355.pdf CORELATION BETWEEN TESTING: INTRADERMAL VS

SEROLOGICAL http://www3.interscience.wiley.com/journal/120745931/abstract?CRETRY=1&SRETRY=0

RUSH IMMUNOTHERAPY: A SHORTENED SCHEDULE http://linkinghub.elsevier.com/retrieve/pii/S1081120610626427

ADDITIONAL SOURCES: VETERINARY CLINICAL IMMUNOLOGY

RICHARD E.W. HALLIWELL PhD DACVD & NEIL T. GORMAN PhD, DACVIM; 1989 MULLER & KIRK’S SMALL ANIMAL DERMATOLOGY, 6TH EDITION

DANNY W. SCOTT, DVM, WILLIAM H. MILLER, JR. VMD., CRAIG E. GRIFFIN, DVM, 2001 THE 5 MINUTE VETERIARY CONSULT CLINICAL COMPANION (SMALL ANIMAL

DERMATOLOGY) KAREN HELTON RHODES, DVM; 2002

LYNDEE KELVER SPECTRUM LABSLKELVER@VETALLERGY.COMF: WWW.FACBOOK.COM/VETALLERGYT: WWW.TWITTER.COM/VETALLERGYYT: WWW.YOUTUBE.COM/VETALLERGYSPOTLI: WWW.LINKEDIN.COM/LYNDEEKELVERB: VETALLERGYBLOG.COM

THANK YOU!