Thyroid Function Test
INTRODUCTION TO
THYROID FUNCTION TEST
By AKANKSHA BHALERAO
Roll no. 51
Anatomy and Physiology of
Thyroid Gland
Anatomy
▪︎Butterfly-shaped endocrine gland.
▪︎Located in front of trachea (neck region)
▪︎Consists of two lobes + isthmus.
▪︎Made up of thyroid follicles.
Physiology
▪Synthesizes hormones:Thyroxine (T4) & Tri-iodothyronine (T3)
(Calcitonin – involved in calcium regulation)
▪Iodine is essential for hormone synthesis.
▪Regulated by:Thyroid Stimulating Hormone.
▪Functions: -Regulates basal metabolic rate (BMR)
-Controls growth and development.
Thyroid Hormones: T3 ,T4
and Calcitonin
Tri-iodothyronine (T3)
• Most biologically active hormone.
• Formed mainly by peripheral conversion of T4
• Rapidly acts on target tissues.
Thyroxine (T4) Calcitonin
• Secreted by parafollicular (C) cells
• Major circulating hormone.
• Decreases blood calcium levels.
• Acts as a prohormone for T3
• Inhibits bone resorption.
• Has longer half-life.
Importance of TFT in
clinical practice
1. Diagnosis of thyroid disorders
~Hypothyroidism
~Hyperthyroidism
2. Differentiation of disorders
~Primary vs secondary thyroid diseases.
~Based on Thyroid Stimulating Hormone levels
3. Monitoring treatment
~Assess response to therapy
~Dose adjustment of drugs.
4. Screening
~Neonatal hypothyroidism
~Pregnancy screening.
5. Evaluation of metabolic status
~Thyroid hormones regulate metabolism
→ TFT reflects metabolic activity
Regulation of Thyroid
Function
by Dipak bhambare
roll no. 52
Hypothalamic-Pituitary-Thyroid (HPT)
Axis
Definition:
A complex neuroendocrine feedback loop that
regulates thyroid hormone production.
Core Function:
It maintains homeostasis by controlling the body's metabolism,
energy expenditure, and growth.
The Three Components:
1. Hypothalamus: The command center in the brain.
2. Pituitary Gland: The master gland that processes and relays the command.
3. Thyroid G he target gland that executes the uction.
TRH and TSH Regulation
The Biochemical Cascade
1. Hypothalamus (The Trigger):
• Secretes TRH (Thyrotropin-Releasing Hormone).
• Action: Travels directly to the anterior pituitary gland to stimulate
it.
2. Pituitary Gland (The Messenger):
Responds to TRH by secreting TSH (Thyroid-Stimulating Hormone).
Action: Enters the bloodstream and binds to receptors on the thyroid
gland.
3. Thyroid Gland (The Producer):
• Stimulated by TSH to synthesize and release thyroid hormones:
Thyroxine (T4) and Triiodothyronine (T3).
The Negative Feedback Mechanism
The HPT axis uses a negative feedback loop to prevent hormonal imbalance
•When T3 and T4 levels are HIGH:
Elevated hormones act on both the hypothalamus and pituitary gland.
They inhibit the release of TRH and TSH.
Result: Thyroid hormone production slows down to restore balance.
•When T3 and T4 levels are LOW:
The inhibitory brake is removed.
The hypothalamus increases TRH, boosting TSH,
which signals the thyroid to produce more hormones.
Indications and Types of
Thyroid Function Test
By Purva Bhoir
Roll no.53
Why TFTs are done?
Thyroid Hormone exerts its action on every cell of body.
It helps in thermogenesis, Regulates Basal Metabolic Rate (BMR)
Thyroid Function Tests are performed to evaluate how well the
thyroid gland is working and to diagnose thyroid disorders.
Main Reasons for Doing TFTs-:
1. Diagnosis of Hypothyroidism (Less secretion of TH)
2. Diagnosis of Hyperthyroidism (Excess secrtion of TH)
3. Monitoring Thyroid Treatment
4. Detection of Autoimmune Thyroid Disorders
5. Evaluation of Goiter or Thyroid Swelling
6. Investigation of Infertility and Menstrual Problems
7. Assessment of Pituitary or Hypothalamic Disorders
Screening and Diagnostic Thyroid
Function Test
Screening Tests Diagnostic Tests
Detect possibility of disease Confirm exact disease
Simple and sensitive More specific
Done first Done after abnormal screening
Example: TSH Example: T₃, T₄, antibodies
Diagnostic Test-
Screening Tests-
a)Total T₃ and Free T₃
a) Serum TSH (Most Important
b) Total T₄ and Free T₄
Screening Test)
c) Thyroid Autoantibody Tests
• Anti-TPO Antibody
b) Free T₄ (FT₄) • Anti-Thyroglobulin Antibody
• TSH Receptor Antibody (TRAb)
c) Neonatal Screening Test d) Thyroid Scan
e) Thyroglobulin Test
Normal Clinical
Test What it Measures Increased In Decreased In
Function/Role Importance
Pituitary hormone
TSH (Thyroid Stimulates T₃ and T₄ Best screening test for
controlling thyroid Hypothyroidism Hyperthyroidism
Stimulating Hormone) secretion thyroid disorders
gland
Total T₃ Total circulating T₃ Active thyroid hormone Hypothyroidism, severe Detects
Hyperthyroidism
(Triiodothyronine) hormone regulating metabolism illness hyperthyroidism
Unbound active T₃ Reflects biologically More accurate than
Free T₃ (FT₃) Early hyperthyroidism Hypothyroidism
hormone active T₃ total T₃
Total circulating T₄ Major hormone Evaluates thyroid
Total T₄ (Thyroxine) Hyperthyroidism Hypothyroidism
hormone produced by thyroid activity
Most important
Unbound active T₄ Indicates true thyroid
Free T₄ (FT₄) Hyperthyroidism Hypothyroidism confirmatory test with
hormone status
TSH
Anti-Thyroglobulin Antibody against Indicates autoimmune Supports autoimmune
Hashimoto thyroiditis Usually absent
Antibody thyroglobulin thyroid damage diagnosis
TSH Receptor Antibody Antibody stimulating Causes excess thyroid Absent in normal Specific for Graves
Graves disease
(TRAb) TSH receptor hormone production individuals disease
Protein produced by Precursor for thyroid Thyroid cancer Tumor marker in thyroid
Thyroglobulin (Tg) After thyroidectomy
thyroid gland hormone synthesis recurrence, thyroiditis cancer
Imaging of thyroid Shows gland structure Toxic nodules, Graves Reduced uptake in Detects nodules and
Thyroid Scan
gland and function disease thyroiditis gland activity
Neonatal TSH Detects congenital Congenital Prevents mental
TSH level in newborns Rare
Screening hypothyroidism hypothyroidism retardation in infants
TSH
THYROID STIMULATING HORMONE
[Link]
54
TSH
TSH is also called thyrotropin
Secreted by the anterior pituitary gland
Regulates growth and function of the thyroid gland
Stimulates synthesis and release of:
Thyroxine (T4)
Triiodothyronine (T3)
Regulation
Controlled by Thyrotropin Releasing Hormone (TRH) from
hypothalamus
TSH secretion is stimulated by TRH
prolactin inhibitory factor (PIF) inhibits secretion of TSH
also controlled by the level of thyroid hormones
Mechanism of Action
TSH binds to TSH receptors on thyroid follicular cells
Activates adenylate cyclase → increases cAMP
Enhances thyroid hormone synthesis
Functions of TSH
Increases iodine uptake
Stimulates thyroglobulin synthesis
Promotes iodination and coupling reactions
Increases release of T3 and T4
Causes growth and vascularity of thyroid gland
Clinical Significance
Increased TSH Seen in:
Primary hypothyroidism
Iodine deficiency
Hashimoto thyroiditis
Decreased TSH Seen in:
Hyperthyroidism
Pituitary insufficiency
Excess thyroid hormone therapy
Laboratory Estimation
Serum TSH measured by immunoassay
Most sensitive test for thyroid dysfunction
Normal Values
Approximately 0.4 – 4.0 mIU/L
Conclusion
TSH is the major regulator of thyroid hormone
synthesis and secretion
Important marker in diagnosis of thyroid disorders
T3-Triiodothyronine
By- Abhinaya (55)
:-Total T3 Includes:
Protein-bound T3 (major portion)
Free T3 (small active portion)
:-Free T3
Unbound form circulating freely in blood
Biologically active hormone
ESTIMATION TECHNIQUES
[Link] (RIA)
Based on antigen-antibody reaction using radioactive labels
Highly sensitive
Previously widely used
[Link] LINKED IMMUNOSORBENT ASSAY (ELISA)
Common modern method
Uses enzyme-labeled antibodies
Color intensity is proportional to hormone concentration
[Link] Immunoassay (CLIA)
Most commonly used automated method
Uses light-emitting chemical reactions
[Link] Dialysis (for Free T3)
Gold standard for FT3 measurement
Separates free hormone from protein-bound
hormone before assay
THYROXINE (T4) TEST
-Ananya(56)
What is T₄?
T₄ (Thyroxine) is the principal hormone secreted by the thyroid
gland.
It is synthesized from the amino acid tyrosine and iodine.
About 90% of thyroid hormone released is T₄.
T₄ acts mainly as a prohormone and is converted into active T₃ in
peripheral tissues by deiodinase enzymes.
Secretion of T₄ is regulated by TSH from anterior pituitary.
Functions of T₄
Increases basal metabolic rate (BMR)
Stimulates oxygen consumption in tissues
Regulates carbohydrate, protein and fat metabolism
Essential for normal growth and skeletal development
Necessary for CNS maturation and mental development
Increases cardiac output and heart rate
Maintains body temperature
Forms of T₄ in Blood
Total T₄:
Includes protein-bound and free hormone
Free T₄ (FT₄):
Physiologically active fraction
Represents thyroid functional status more accurately
Normal Values:
Total T₄ : 5–12 µg/dL
Free T₄ : 0.8–2.0 ng/dL
(Reference values vary among laboratories)
– T₄ Test and Methods of Estimation
What is the T₄ Test?
T₄ test measures concentration of thyroxine hormone in serum.
It is an important investigation in assessment of thyroid
function.
Indications of T₄ Test
Suspected hypothyroidism
Suspected hyperthyroidism
Goiter evaluation
Monitoring thyroid hormone therapy
Assessment of pituitary-thyroid axis
Screening in congenital hypot
Clinical Interpretation of T₄ Levels :
Increased T₄ Levels – Hyperthyroidism
Graves’ disease
Toxic multinodular goiter
Thyroid adenoma
CAUSES
Excess administration of thyroxine
Thyroiditis in early phase
CLINICAL:
Weight loss despite increased appetite
Heat intolerance
Excess sweating
Nervousness and tremors
Tachycardia and palpitations
Exophthalmos in Graves’ disease
Decreased T₄ Levels – Hypothyroidism
Causes
Hashimoto thyroiditis
Iodine deficiency
Pituitary or hypothalamic disorders
Thyroidectomy
Congenital hypothyroidism
Clinical Features
Fatigue and lethargy
Weight gain
Cold intolerance
Dry coarse skin
Bradycardia
Mental slowing and constipation
Clinical Importance and Conclusion
Importance of T₄ Test
Major component of Thyroid Function Tests (TFTs)
Helps in diagnosis of thyroid disorders
Useful in monitoring treatment response
Assists in evaluation of pituitary-thyroid axis
Important in neonatal screening programs
Factors Affecting T₄ Levels
Pregnancy increases thyroid binding globulin (TBG)
Liver diseases alter protein binding
Severe illness may alter thyroid profile
Thyroid
autoantibody
By - Chinky (57)
Thyroid Antibodies
• Thyroid antibodies are autoantibodies produced against thyroid
gland
components
• They are mainly associated with autoimmune thyroid diseases.
• These antibodies may destroy thyroid tissue or stimulate
excess thyroid hormone production.
• Major thyroid antibodies include:
Anti-Thyroid Peroxidase Antibody (Anti-TPO)
• Anti-Thyroglobulin Antibody (Anti-Tg)
• TSH Receptor Antibody (TRAb)
Anti-Thyroid Peroxidase Antibody (Anti-TPO)
• Most common thyroid antibody
• Directed against thyroid peroxidase enzyme required for T3 and T4
synthesis.
• Commonly present in Hashimoto thyroiditis and Graves disease
Anti-Thyroglobulin and TSH Receptor Antibody
• Antibody against thyroglobulin protein of thyroid follicles.
• Seen in Hashimoto thyroiditis and some thyroid cancers.
• Less sensitive compared to Anti-TPO antibody.
TSH Receptor Antibody (TRAb)
• Directed against TSH receptor on thyroid cells.
• Stimulates thyroid gland continuously.
• Causes excessive production of T3 and T4 leading to
hyperthyroidism.
• Strongly associated with Graves disease.
Effects of Thyroid Antibodies
• Inflammation of thyroid gland
• Destruction of thyroid tissue
• Hypothyroidism or hyperthyroidism
Other methods of estimation
By - Vinanti (58)
T3 is an important thyroid hormone
produced mainly by the thyroid gland and
partly by conversion of T4 in peripheral
tissues
regulates metabolism, growth, development, and energy production.
1. Thyroglobulin
Glycoprotein produced by thyroid
follicular cells
Stored in thyroid colloid and acts as
precursor for T3 and T4 synthesis
Small amount normally released into
blood
Marker for monitoring differentiated
thyroid cancer after thyroidectomy
[Link]-Binding Globulin (TBG)
Main plasma protein that transports thyroid
hormones in blood
Binds most circulating T3 and T4
Alters total T3/T4 levels without changing free
hormone levels
Important in interpretation of thyroid function
tests
3. Reverse T3
Inactive form of T3 produced from peripheral
conversion of T4
Formed mainly during stress or illness
Regulates metabolism during stress by reducing
metabolic activity
[Link] / Radioactive Iodine Uptake (RAIU)
Evaluates thyroid gland function and iodine
uptake ability
Thyroid gland takes up radioactive iodine
Uptake measured using gamma
camera/scanner
Functional assessment of thyroid gland
Helps guide treatment planning
Interpretation of TFT
Patterns
By Deewanshi (59)
Abnormalities of Thyroid function
Feature Hyperthyroidism Hypothyroidism
Excess secretion of throid Decreased secretion of
Definition
hormones thyroid hormones
Grave's disease,toxic
multinodular goiter, Autoimmune
functioning metastaic throiditis(Hashimoto's
Common causes
thyroid causes thyroiditis),thyroidectomy,
carcinoma,iodine excess lithium, antithyroid drugs
Metabolic rate Increased Decreased
Body weight Weight loss Weight gain
Heat tolerance Sensitive to heat Cold intolerance
Feature Hyperthyroidism Hypothyroidism
Cardiovascular effects Tachycardia,Palpitations Slow heart rate
Anxiety, emotional disturbances ,
Nervous system effects Sluggishness, slow responses
tremors
Plasma T3/ T4 Increased Decreased
Plasma TSH Decreased Increased
Often diffuse goiter in Grave's May be enlarged in
Throid gland
disease autoimmune thyroiditis
Primary hypothyroidism vs Secondary
hypothyroidism
Feature Primary hypothyroidism Secondary hypothyroidism
Pituitary gland (decreased TSH
secretion)
1. Site of defect Thyroid Gland
Hypothalamus (decreased TRH
secretion)
[Link] Level Increased Decreased
4.T3 and T4 Level Decreased Increased
[Link] gland Normal or enlarged (goiter may be present) Norma (not enlarged)l
Feature Primary hypothyroidism Secondary hypothyroidism
Common in adults (especially
[Link] Can occur at any age
women)
Typical features of
Similar to primary but may be
hypothyroidism (lethargy, weight
[Link] milder ;other pituitary
gain , cold intolerance,
deficiencies may be present )
constipation)
Increased TSH with decreased Decreased TSH (or normal) with
[Link]
T3/T4 decreased T3/T4
Levothyroxine (Thyroid hormone Treat underlying cause and
[Link]
replacement) Levothyroxine (if needed)
Clinical Correlation and
Conclusion
By Dev
Roll no. 60
Factors affecting TFT results
Factor Effect on TFT Results
Newborns and elderly people may have different
Age
normal thyroid hormone levels
Pregnancy Increased TBG causes increased total T3 and T4
Can alter TSH and thyroid hormone levels (sick
Stress / Acute illness
euthyroid syndrome)
Steroids, dopamine, lithium, amiodarone, oral
Drugs/Medications
contraceptives, anticonvulsants etc. affect TFTs
Excess or deficiency alters thyroid hormone
Iodine intake
synthesis
Abnormal TSH secretion affects thyroid hormone
Pituitary or hypothalamic disorders
levels
Different assays may produce slightly different
Laboratory method variations
values
Presence of thyroid antibodies affects thyroid
Autoimmune diseases
function
GOITER
[Link]: Goitre is an abnormal enlargement of
the thyroid gland present in the neck
[Link] of Goitre: [Link] deficiency
[Link] thyroiditis
[Link] nodules
[Link] tumors
[Link] Features:[Link] in front of neck
[Link] in swallowing
[Link] in breathing
[Link] of voice
[Link] of hyperthyroidism or
hypothyroidism
[Link]:aThyroid Function Tests (T3, T4, TSH)
[Link] of thyroid
[Link] scan
dThyroid antibody tests
[Link]: [Link] supplementation
[Link] therapy
[Link] removal of thyroid
(thyroidectomy)
[Link]:Adequate iodine intake through iodized
salt and balanced diet helps prevent endemic goitre.
GRAVE’S DISEASE
[Link]:Graves’ disease is an autoimmune disorder causing
hyperthyroidism due to excessive production of thyroid
hormones.
[Link]: production of thyroid-stimulating immunoglobulins
(TSI) that continuously stimulate the TSH receptors.
[Link] Factors: [Link] sex
[Link] history
c. Stress
[Link]
[Link] disorders
[Link] Features:[Link] toxic goitre
[Link] loss despite increased appetite
[Link] intolerance
[Link] sweating
[Link] Manifestations (Exophthalmos): Bulging of eyes due to
autoimmune inflammation behind the orbit.
[Link] Findings: [Link] T3 and T4
[Link] TSH
[Link] thyroid antibodies
[Link]: a,Antithyroid drugs
[Link] blockers for symptom relief
[Link] and Care:Early diagnosis, regular follow-up, stress
management
Plasma Total T3 & FT3 & FT4 in
Condition Plasma TSH Response to TRH
T4 Plasma
Graves’ disease Increased Increased Decreased Nil
Toxic goiter Increased Increased Decreased Nil
T3 increased, T4 FT3 increased, FT4
T3 toxicosis Decreased Sluggish
normal normal
Excess intake of FT3 increased, FT4
Increased Decreased Sluggish
thyroxine normal
Takeaway points-
The main TFT parameters include TSH, T₃, and T₄, which help diagnose hypothyroidism and hyperthyroidism.
TSH is the most sensitive screening test, while Free T₃ and Free T₄ provide accurate assessment of thyroid hormone
activity.
Additional tests like thyroid antibodies, thyroglobulin, and RAIU help identify autoimmune thyroid diseases and other
thyroid disorders.
TFTs are important for diagnosis, treatment monitoring, neonatal screening, and evaluation of metabolic and hormonal
health.
THANK YOU !
For your attention.