Antibody-mediated Immunity Immunology Unit Department of Pathology College of Medicine, KSU.
College of Medicine, KSU Medical education Department Pathology Department
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Transcript of College of Medicine, KSU Medical education Department Pathology Department
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College of Medicine, KSUMedical education Department
Pathology DepartmentMedical Biochemistry Unit
GIT Block (2nd Year)Integrated Practical (Biochemistry / Pathology)
Liver Function Tests
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Measurement of total bilirubin
ByMedical Biochemistry Unit
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Q1. What are the liver function tests (LFTs)?
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Q1. What are the liver function tests (LFTs)?
e.g. Viral hepatitis(AST)
(ALT)
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Q2. What is bilirubin and how is it produced in the body?
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Q2. What is bilirubin and how is it produced in the body?
• Bilirubin is a yellow bile pigment. • It is produced from the degradation of heme;
which is one of the breakdown products of red blood cells.
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Q3. Which form of bilirubin is carried to the liver and how?
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Q3. Which form of bilirubin is carried to the liver and how?
• The unconjugated form of bilirubin is carried to the liver
• Unconjugated bilirubin forms a complex with albumin to be transported
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Q4.•How & why is bilirubin conjugated?•On the picture below, mark the intracellular location for the process of conjugation?•Mention 2 syndromes due to congenital deficiency of the conjugating enzyme (bilirubin glucuronyl-transferase).
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How & why is bilirubin conjugated?
• Bilirubin is conjugated by binding to glucuronic acid in hepatocytes
• The conjugated-bilirubin is water soluble and can be excreted in the urine and feces
• This prevents precipitation and deposition in tissues.
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Defective enzymatic conjugation of bilirubin
• Examples of clinical conditions due to congenital deficiency of the conjugating enzyme ( bilirubin glucuronyl transferase)– Crigler-Najjar syndrome– Gilbert syndrome
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A. How is bilirubin eliminated from the body?
B. What are the fates of bilirubin in the intestine?
Q5.
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1
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5
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Blank7
Using blank setthe base line at
wavelength 578 nm
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Read test (A) at 578 nm
Measurement of Total Bilirubin
3
Add 200µl Reagent 1
Add 50µl Reagent 2
Add 1 ml Reagent 3
Blank Test
Add 200µl serum
Add 1 ml Reagent 4
Test
Only in test
4
To blank and test
To blank and test
To blank and test
Transfer(Pour) to cuvette
Mix and wait for 5-10 min at RT
Mix and wait for 5-20 min at RT
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Calculation of total bilirubin concentration
Conc. of serum total bilirubin: A × 185 = …... µmol/L
Note- (Normal range: 2 – 17 µmol/L)
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Group B Test 1 Test 2
B1B2B3B4B5B6B7Mean
Estimation of total Bilirubin
Group Test 1 Test 2
A1
A2
A3
A4
A5
A6
A7
Mean
Group C Test 1 Test 2
C1
C2
C3
C4
C5
C6
C7
Mean
With Dr. Ahlam With Dr. Reem With Dr. Sumbul
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Thankyou