allergies overview
TRANSCRIPT
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 [email protected]: WWW.FACBOOK.COM/VETALLERGYT: WWW.TWITTER.COM/VETALLERGYYT: WWW.YOUTUBE.COM/VETALLERGYSPOTLI: WWW.LINKEDIN.COM/LYNDEEKELVERB: VETALLERGYBLOG.COM
THANK YOU!