DIAGNOSTICS OF

ALLERGIES
Immunology
Paola Viñé Ortega
WHAT IS AN ALLERGY?
Allergies are caused by an abnormal response of the immune system. The immune system
reacts to a usually harmless substance in the environment. This substance can be pollen, mold,
dust, animal dander, certain foods, insect stings, etc. and is referred to as an allergen.

The term allergy is used to identify hypersensitivity reactions caused by the excessive
degranulation of mast cells and basophils induced by a specific allergen, against which a
hypersensitive individual produces IgE antibodies.
CURRENT SITUATION
The World Allergy Organisation (WAO)
estimate of allergy prevalence of the
whole population by country ranges
between 10 - 40%.

• Allergy is a very common ailment,

affecting more than 20% of the
populations of most developed
countries.

• Allergic disease is the 5th leading

chronic disease among all ages.

• Allergies are the 3rd common chronic

disease among children under 18 years
old; up to one child in three is affected.
CURRENT SITUATION
• Allergy is the most common chronic disease in Europe. Up to 20% of patients with
allergies live with a severe debilitating form of their condition, and struggle daily with the
fear of a possible asthma attack, anaphylactic shock, or even death from an allergic
reaction.

• More than 150 million Europeans suffer from chronic allergic diseases and the current
prediction is that by 2025 half of the entire EU population will be affected

• Seven times as many people were admitted to hospital with severe allergic reactions in
Europe in 2015 than in 2005.

• The avoidable indirect costs of failure to properly treat allergy in the EU is estimated to
range between 55 and 151 billion Euro per annum.
WHEN TO PERFORM ALLERGY DIAGNOSTIC
● Assist in confirming or excluding IgE sensitization in support of a clinical
history based diagnosis of stinging insect hypersensitivity, drug allergy, food
allergy or aeroallergen allergy.
● Determine the need for environmental control recommendations to reduce
exposure to outdoor or indoor aeroallergens.
● Demonstrate sensitization to inhalant occupational allergens, which may
cause occupational asthma or rhinitis
● Guide the selection of aeroallergens for inclusion in allergen immunotherapy
extracts
ALLERGY ASSESSMENT
SKIN TEST
LABORATORY ASSESSMENTS
SPIROMETRY
THE BRONCHODILATION TEST
THE BRONCHOCONSTRICTION TEST
IMAGING TEST
EXPOSURE AND ELIMINATION TEST
PHYSICAL TEST

The main goal of allergy diagnostics is the identification of causative allergens. Therefore,
the proper diagnosis of allergies should start with detailed anamnesis focusing on the
assessment of the history and nature of symptoms and the analysis of triggering factors.
DIAGNOSING ALLERGIES
Allergists are experts in their field with specialized training that allows them to:
● Perform allergy testing
● Identify the source of your suffering
● Accurately diagnose your condition
● Treat more than just your symptoms
● Develop a personalized plan that eliminates your symptoms
● Provide you with the most cost-effective care that produces the best results

Two key steps in the process of allergy diagnosis are the medical history and allergy test
selection. Allergists use their skills in these areas to help more patients feel well, stay
active during the day, and rest at night. And that’s nothing to sneeze at.
SKIN TEST
PRICK TEST

Skin test is the most sensitive and accurate method to

diagnose allergic sensitization.
The test requires the choice of tested allergens and the
application of positive and negative controls.

PROCESS:
● Prick test is performed on the volar forearm
● Prick test involves the injection of allergen extract into
the skin (epidermis),resulting in its binding to the
membrane IgE of mast cells, causing their
degranulation.
● The release of histamine and other vasoactive
substances from mast cells results in individuals
sensitive to the particular allergen in erythema and
induration with pruritus at the application site within 20
minutes of application.
SKIN TEST
PRICK TO PRICK TEST

The Prick to Prick test is a modification of the standard Prick test

used almost exclusively in the diagnosis of food allergy.
Food allergy is often masked by the cross-reactivity of allergens.

PROCESS
● Prick to prick is performed on the volar forearm as the
prick test.
● The needle being first inserted in the fruit or vegetable,
trying to have a minimum quantity of tested food on the
needle tip.
● The needle is then applied directly to the skin to test for the
food allergen injected to the mast cells in the skin. The skin
reaction, wheal and erythema, is assessed also after 20
minutes.
SKIN TEST
INTRADERMAL SKIN TEST

Intradermal skin test are exclusively for

the diagnosis of drug allergy, requiring
only the use of injectable forms of drugs.
Tests used in the assessment of specific
immune responses, adverse drug
reaction or photo-toxicity and
immunopathological mechanisms.

Intracutaneous testing is used in hospitalized patients on a strict diet are most commonly applied
with an intracutaneous placebo solution (saline solution) and 5 cm from it the tested solution, test
performed in the area between the neck and shoulder.

The reaction is assessed after 15 minutes and then 8 hours after being administered, being allowed
the administration on another part of the back the next day.
SKIN TEST
EPICUTANEOUS “PATCH” TEST
● These tests are used to demonstrate type IV hypersensitivity,
which is clinically important in allergic contact dermatitis.
● This hypersensitivity results from sensitization by different
chemical compounds, such as metals, components of perfumes
and cosmetics, rubber components and certain drugs,
formaldehyde, lanolin and others. This test is possible to
perform through the skin but also orally.
● This diagnostic test consists of applying different substances to
the skin of the back on supports called patches, similar to
stickers, and on top of these is applied tape.
● These substances and tapes are usually left for 2 days. At 48
hours they are removed and 1 hour later, the first reading is
performed which consists of looking at the back and
identifying if there is any positive reaction. The second reading
takes place at 96 hours. Occasionally a late reading is done
after 7 days.
IS ALLERGY SKIN TESTING SAFE?
Skin testing is extremely safe, especially when performed by an allergist experienced in the diagnosis
of allergies. Whole-body allergic reactions, sometimes called anaphylaxis, are extremely rare from
skin testing.

Certain groups of people cannot have skin testing, and therefore allergy blood testing is a better test.
These groups include those who cannot stop their antihistamine medications; those with sensitive
skin, those taking certain blood pressure medications, those with serious heart and lung conditions
that put them at increased risk if anaphylaxis should occur.
LABORATORY ASSESSMENTS
Allergic disorders are often accompanied by laboratory markers such as eosinophilia.

The highest levels of total IgE may be observed in atopic eczema, the total IgE are
diagnosed using methods that are capable of determining the levels of specific IgE.

If IgE antibody are present in patient’s plasma, the patient is with high probability
clinically hypersensitive to the tested allergen.

The sIgE assessment is indicated in the case of sensitization against an allergen that is not
included in the skin diagnostic set, being assessed by non-radio modifications of
immunoassays such those where the allergen is bound to a solid phase.
BASOPHIL ACTIVATION TEST
Basophils are white blood cells from the bone marrow that play a
role in keeping the immune system functioning correctly. Basophils
are the biggest stores of soluble mediators in blood circulation

Basophils are capable of respond to the allergen , expressing a

specific IgE antibody in their membrane against a particular allergen
are exposed to its action. The result is called degranulation, which is
possible to detect in an optical microscope profiting from the fact of
the affinity of this granules to basic stains such as toluidine blue.

The results of the test may be available as soon as 4 hours.

BASOPHIL ACTIVATION TEST

Basophil activation can be express by CD63 or CD203c, used in Flow-CAST to determine the number
of activated basophils using flow cytometry to quantify the number of activated basophils in proportion
to their total number.
It may be used in the diagnosis of allergy to certain foods and drugs such as muscle relaxants B-lactam
antibiotics or insect venom in anaphylactic or non-sensitized patients.
The test of lymphocyte transformation assay is occasionally used in the diagnosis of drug allergies.
SPIROMETRY
Diagnostic spirometry is used to assess a patient’s lung
function for purpose of comparison with a normal
population, or with previous measures from the same
patient.

Spirometry is essential for the successful diagnosis of

bronchial asthma, as well as for monitoring treatment
success. This assessment of lung function using the
spirometry device is used to measure lung volume and
flow rates. In order to determine the degree of bronchial
obstruction used to classify the degree of the disorder,
forced vital capacity

(FVC) and forced expiratory volume in 1 second

(FEV1) are used.
THE BRONCHODILATION TEST
● An initial spirometry is performed to assess the
patient's native respiratory status.
● In the bronchodilation test in patients with
reduced FEV1, a bronchodilator is applied and
the subsequent measurement assesses the
reversibility of the bronchial obstruction. You
will wait about 15 minutes and then the
spirometry is repeated.
● An increase in FEV1 of >200 ml is considered
a positive result.
THE BRONCHOCONSTRICTION TEST
● In the bronchoconstriction test, a
bronchoconstrictor (histamine or methacholine)
is applied to a patient with normal FEV1 value.

ALLERGEN- INDUCED CONSTRICTION

● Inhalation of allergens in sensitized subjects
develops into bronchoconstriction within 10
minutes, reaches a maximum within 30 minutes,
and usually resolves itself within one to three
hours. In some subjects, the constriction does
not return to normal, and recurs after three to
four hours, which may last up to a day or more.
The first is named the early asthmatic response,
and the latter the late asthmatic response.
IMAGING METHODS
Imaging is not commonly used for diagnosing allergies. However, an X-ray of your lungs
or sinuses may be done to rule out other conditions. Studies such as a sinus computed
tomography (CT) scan might be used if chronic sinusitis is suspected.

X-ray assessment of paranasal sinuses capable of identifying chronic inflammatory

changes, and other causes of nasal obstruction and discharge may be excluded.

EXPOSURE AND ELIMINATION TEST

Is the most sensitive diagnostic method for the assessment of food allergies.

They are based on elimination diet (e.g.rice) and subsequent alternative oral
administration of foods and placebo. Is necessary to eliminate the subjective feelings or
subjective bias of the patient.
PHYSICAL TEST
A suitable physical test is selected on the basis of history, in order to diagnose the allergy.

A patient with a suspected allergy is exposed for a certain time to physical effects.

Later, we observe erythematous symptoms or urticarial on a patient’s skin.

Physical test are probed by a ways such as frozen ice test tubes and warm objects.