Total S.Bilirubin
Bilirubin: Bilirubin is a yellow-orange compound that forms in the body as a result of the breakdown of hemoglobin from red blood cells. Hemoglobin is the protein responsible for carrying oxygen in red blood cells, and as these cells age and are broken down, bilirubin is produced as a byproduct.
Bilirubin is transported to the liver, where it undergoes further processing to become a water-soluble form that can be excreted from the body. This processed bilirubin is then excreted into the bile, which is a digestive fluid produced by the liver. From the bile, bilirubin eventually enters the intestines and is eliminated from the body in the stool. The color of bilirubin gives a yellowish tint to both bile and stool. When there is an issue with the liver's ability to process bilirubin, or when there is an excessive breakdown of red blood cells, bilirubin levels in the bloodstream can rise, leading to jaundice, which is the yellowing of the skin, sclera, and mucous membranes. Elevated bilirubin levels can be a sign of various medical conditions, and they are typically measured through blood tests to help diagnose and monitor these conditions.
Jaundice:
Jaundice, also known as icterus, is a medical condition characterized by the yellowing of the skin, sclera (the white part of the eyes), and mucous membranes. This discoloration occurs when there is an accumulation of free or conjugated bilirubin in the bloodstream. Jaundice becomes clinically noticeable when the total plasma bilirubin levels exceed 2 mg/dL (34 µmol/L).
Principle: Enzyme solution + serum ( bilirubin ) -color complex.
The intensity of the color is proportional to the amount of glucose which is measured by a colorimeter.
Classification Of Bilirubin and Jaundice Test Name: There are Three Types
1. Total Bilirubin
2. Direct Bilirubin
3. Indirect Bilirubin
Classification Of Jaundice: There are three types
1. Hemolytic or pre-hepatic
2. Hepatocellular or hepatic
3. Obstructive or cholestatic or post hepatic
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| Total Bilirubin Test |
A. Hemolytic jaundice
1. Hemolytic anemia like Malaria, Eryth elastosis, fetalis, Thalassemia, etc.
2. Increase production of shunt bilirubin in the bone
3. Massive hemorrhage within the body.
B. Hepatocellular jaundice:
1. Reduced uptake of bilirubin by hepatic cells: Gilbert's disease, drug-induced etc.
2. Impaired conjugation of bilirubin: Neonatal jaundice, Crigier-Naji syndrome, Gilbert's disease.
3. Impaired excretion of conjugated bilirubin
a. Intrahepatic causes: Dubin-Johnson syndrome, Rotor syndrome, drug-induced (estrogen, steroid), etc.
b. Extra hepatic causes: It occurs due to the obstruction of the biliary tract. The obstruction may be in the lumen, in the wall, or outside of the tract:
i. Lumen: Gallstone, Worm, etc.
ii. Wall: Cholangitis, Tumor, Atresia, Stricture, etc.
iii. Outside the wall: Ca of the head of the pancreas, Sub-hepatic abscess, Enlarged lymph node, etc. Combination of all viral hepatitis, Septicemia,
4. Cirrhosis of the liver, Hepatic poisoning, etc. jaundice
C. Obstructive jaundice: Same as the intra and extra-hepatic causes of impaired excretion of bilirubin in hepatocellular jaundice.
Principle of Bilirubin : Enzyme solution + serum ( bilirubin ) = color complex
The intensity of the color is proportional to the amount of bilirubin which is measured by a colorimeter
Sample Collection Procedure:
- The Vials should be checked for appropriate kinds and for barcode or paper labeling
- Put the patient in a comfortable position with the arm properly supported.
- Apply a tourniquet above the elbow joint
- Ask the patients to make a fist S.
- Think of four things when feeling for a vein bounce, direction of the vein, size of the needle, and depth.
- They are cleaned with an area of 70%: ethanol and allowed to dry in the air.
- Check the needle, especially the tip, and check for any blockage by pressing the piston.
- Grasp the back of the forearm and make the overlying skin tight.
- The vein is penetrated by positioning the needle 15, a degree angle to have Red Hat.
- Draw and piston slowly.
- After blood has been drawn the patients should release the first and the tourniquet is also removed.
- Press the vein puncture area with the cotton with antiseptic as soon as the needle is removed. The patients may remove the cotton after 7-10 minutes.
- After removing the needle the collected blood is dispensed in the appropriate tubes.
- After vein puncture, the needle should be removed from the syringe and disposed of by using a needle destroyer. Reagent Preparation: Reagent and standard are ready for use. Test Procedure: Sample 1000 ml Standard
Reagent Preparation:
The reagent and standard are ready for use.
Reagent 1- Sample blank reagent
Reagent 2- DCA
Reagent 3- Nitrite Acid
Preparation of working reagent: To prepare 1 ml working reagent, 500 µ l reagent 2 is mixed with 500 µ l reagent 3. The mixture is kept for 15 minutes in the dark before us.
Working reagent Stability Of 10 And in In And in days to have dark of the Red Hat 15-25 ° C, and to have 21 And And 28 And days of the Red Hat ° C.
T.Bilirubin:
Analyzer Parameter:
- Wavelength / Filter : 530 nm, 546 nm
- Method : End Point
- Cuvette : 1cm light path
- Reaction Direction : 30 minutes
- Bank : Blank Reagent
- Reagent Volume : 1000 µ l / 1 ml
- Sample Volume (ul) : 100 µ l
- Normal Low (mg / dl) : 0.2 mg / dl
- Normal High (mg / dl) : 1.4 mg / dl
- Linearity Low (mg / dl) : 0.1 mg / dl
- Linearity High (mg / dl) : 30 mg / dl
- Units : mg / dl
- Temperature : 37 ° C,,
Materials:
- Cotton
- Hexaso
- Tourniquet
- Gloves
- Marker Pen
- Test tube rack
- Disposable Syringe
- Gel tube / Yellow vial
- Stop Watch
- Calculator
- Colorimeter
- Semi-Auto analyzer
- Icubotot
- Micropipette
Test Procedure:
Sample / Test Reagent
Blank 1000 µl -
Working reagent - 1000 µl
Sample 100 ml 100 ml
Mix and keep at room temperature for 10 minutes and 37 ° C for 5 minutes in a dark room. standard and sample should be determined by adjusting 0 with reagent blank
Calculation: Concentration of sample = Factor × OD of sample
= 12.5 × OD of sample (if using 546 nm)
= 17.5 × OD of sample (if using 530 mm)
Normal Value :
Adult: 0.2-1.4 mg/dl
Newborns: up to 13.3 mg/dl
Direct bilirubin Test Procedure :
All reagents are ready for use
Reagent Blank Standard Sample
Reagent-1 2.5 ml 2.5 ml 2.5 ml
Reagent- 2 1 drop 1 drop 1 drop
Standard × 100 ml ×
Sample × × 100 ml
Mix and keep at room temperature for 5 minutes. standard and sample should be determined by adjusting 0 with reagent blank.
Calculation: Concentration of direct bilirubin = OD of Sample / OD of Sample * Concentration of standard
Normal Value: Up to 0.20 mg/dl
Linearity: Up to 15 mg/dl
Caution:
blood sugar is an important test be care must be taken when testing blood sugar to avoid hemorrhagic blood you have to do QC every day as well as run standard
A serum bilirubin test is a common blood test used to measure the levels of bilirubin in the bloodstream. This test is important for diagnosing various medical conditions, particularly those related to liver and gallbladder function, as well as the breakdown of red blood cells. Here are some cautions and considerations related to the serum bilirubin test:
1. Interpretation Requires Context: Interpreting serum bilirubin levels should be done in the context of a person's overall health and medical history. High or low levels of bilirubin can indicate different conditions, and the significance of the results depends on other clinical factors.
2. Medications and Supplements: Inform your healthcare provider about any medications, supplements, or herbal remedies you are taking, as some of these substances can affect bilirubin levels. Certain medications may increase or decrease bilirubin production or interfere with the liver's ability to process bilirubin.
3. Fasting Requirements: In most cases, a serum bilirubin test does not require fasting.
4. Clinical Symptoms: Elevated bilirubin levels alone may not always indicate a problem. Clinical symptoms, such as jaundice (yellowing of the skin and eyes), dark urine, abdominal pain, and fatigue, should also be considered in conjunction with test results.
5. Follow-up Tests: Depending on the results of the serum bilirubin test and other clinical factors, your healthcare provider may recommend follow-up tests or imaging studies to determine the underlying cause of bilirubin abnormalities.
6. Liver Function Assessment: The serum bilirubin test is just one part of assessing liver function. It is often used alongside other liver function tests, such as liver enzyme tests, to gain a more comprehensive understanding of liver health.
7. Pediatric Considerations: In newborns, a high level of unconjugated (indirect) bilirubin can lead to jaundice. Pediatricians routinely monitor bilirubin levels in newborns and may recommend treatment if levels become too high to prevent complications.
Reagent company name:
1. Tradsworth Ltd (Human Gmbh, Germany)
2. Genetic Trading
3. Medi-Vision Ltd
4. Bio-Trade International
5. Bio-Medical Limited
6. Meditech
7. Health Care Technologies
8. Rajanigandha International
9. Randox
10. Plasmatex
11. Leiz Supply Agency
12. Abc International
13. Rose International
14. Biozen.
