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Clinical Biochemistry
PLC 402
➢Prof. Hend Hussein
➢ Ass. Dr. Samer Zahran
Course Assistant:
Ass. Lect. Sara El-Hadey
Clinical Biochemistry course provide an advanced
knowledge of human body chemistry levels under
healthy and abnormal conditions. Also, it evaluates
the suitability of clinical specimens and the
laboratory test outcomes, and explain the relation
between the diseases and the abnormality in blood
chemistry.
Clinical Biochemistry (PLC 402) involves the
study of the proper sampling techniques, study of
chemical components of the human body, plasma
proteins, kidney, liver function, as well as the
study of body electrolytes, acid-base balance,
hormones and disorders in thyroid function.
1. Introduction
2. Blood Sampling
3. Acid-Base balance
4. Electrolytes
5. Kidney Function
6. Acute Phase Protein
7. Hormones 1
8. Hormones 2
9. Thyroid function
10. Diabetes Mellitus
11. Liver Function
12. Cardiac Function
13. Tumor Markers 1
14. Tumor Markers 2
➢ To diagnose diseases
➢ To screen for risk factors
➢ To monitor the effects of treatments &
medications.
In blood tests, blood sample is usually obtained
from the patient's vein. Other specialized blood
tests, such as the blood gases, require blood
extracted from an artery.
➢ Blood gas analysis is used to monitor CO2,
O2, blood pH and bicarbonate levels related
to pulmonary function or certain metabolic
conditions.
Laboratory analysis
Biochemical
analysis
Molecular
analysis
Cellular
analysis
Sodium, potassium, chloride, calcium, phosphate,
magnesium, iron, urea, creatinine, liver enzymes,
copper, zinc, glucose, cholesterol, bicarbonate,
lactate, and others.
➢ Protein electrophoresis
➢ RNA
➢ DNA
DNA testing (via PCR technique) is used in
diagnoses of many disorders and forensic
sciences.
• Complete blood count (CBC)
• Erythrocytes sedimentation rate (ESR)
• Cross-matching (for blood transfusion or
transplantation)
• Blood culture
Universal precautions should be used to increase the
safety of blood collection.
These precautions are important to reduce the risk of
exposure skin and/or mucus membranes to infectious
materials. It includes the use of a variety of protective
barriers, such as gloves, masks, gowns (or white coat),
and eyewear.
Precautions
Universal precautions were designed to prevent the
transmission of HCV, HBV, HIV, and other blood
pathogens.
Universal precautions are applied in blood, semen,
vaginal secretions, pleural fluids, cerebrospinal,
synovial, peritoneal, pericardial, amniotic fluids
and tissues.
universal precautions
You must avoid blood drawing from:
➢ arm affected by a stroke or neurological injury
➢ arm of a mastectomy
➢ areas with extensive scar tissues, shunt, varicose
veins, and the palm
➢ arm of a patients that receive intravenous fluids.
➢ site that is swollen or affected by certain skin condition such
as eczema or other infection
Before the blood drawing, you should ask the
patient about some factors that may affect test
results: medications, pregnancy and age.
Make sure the patient has followed special
instructions such as fasting or taking a
medication.
The patient should be instructed about what blood
tests have been ordered and why.
Ask the patient if anytime has had trouble during the
blood drawing. This question may be an alert to
potential problems in finding an appropriate vein.
Also ask the patients what position would be most
comfortable for them when drawing the specimen.
Venipuncture makes many patients nervous, and
having them lie down.
Physician
Pharmacists
Technicians
Practitioners
paramedics,
Phlebotomists
Nurses staff
Respiratory therapists
Are responsible for blood extraction.
➢ Needles: vacutainer needles, syringes, butterfly needles
➢ Antiseptic. 70% alcohol wipes or disinfection swabs
➢ Tubes
➢ Vacutainer holder
➢Tourniquet (is used to find a vein)
➢Adhesive dressing
➢ Rubber gloves
➢ Pillow or other support
➢ Sharps disposal container
➢ Disposal box
The vacuum tubes are designed to draw a determined volume of
blood. This system consists of vacuumed specimen tubes, a
needle, and a plastic holder. When the tube is placed into the
holder and pressed against the needle, negative pressure results,
and blood is pulled into the tube
➢ Transfer and storage tubes
➢Pipettes
➢ Centrifuge
➢ Racks for tubes
➢ Special boxes for tube transfer and storage
➢ Freezer (for storage)
1. Venipuncture is useful to obtain cells and
extracellular fluids (plasma or serum) from the body
for analysis.
2. Fingerstick : If only a few drops of blood are needed
(make a small prick in the tip of the finger and then
squeeze the blood out). Ex: blood group
3. Heel-stick is the preferred method of blood
collection on a newborn baby or infant
During blood collection, the arm should rest on a
pillow or other supportive prop. The patient open
and close the fist. The phlebotomist sets the
tourniquet around the upper arm, searches the
proper vein by inspecting and palpating and then
sterilizes the injection site. The use of a tourniquet
should be minimized (to avoid changes in blood
hemoconcentration).
After that, the needle beveled upward, should be
pushed smoothly and quickly into the vein, to
minimize the possibility of hemolysis as a result of
vascular damage. Immediately after the insertion,
the tourniquet should be released (to minimize the
effect of hemoconcentration).
The needle will be attached either to a low pressure
blood test bottle (Vacutainer), or to a syringe where
the plunger is pulled back to create low pressure.
When the necessary amount of blood has been
extracted, the needle is removed and a little ball of
cotton wool is held over the wound for 1 minute.
The artery blood it is usually extracted from the
wrist where the artery is very close to the skin.
After taking blood from an artery, a ball of
cotton wool is held over the wound about 5
minutes to stop any bleeding.
Blood collection tubes come with a variety of colored
stopper caps, and may contain additives. The colored
stopper is important to avoid the cross-
contamination of additives between tubes
1. Serum separator tube (SST). This tube contains
a polymer gel for blood chemistry (Glucose, urea,
cholesterol, Na, K, Mg Zn, …..
2. Heparin tubes (green top tubes) are used for lithium
and ammonia levels.
3. EDTA tubes ( with lavender top). For complete
blood counts.
4. Sodium citrate tubes (light blue top) are used for
prothrombin time (PT) and partial prothrombin
time (PTT) analysis.
5. Sodium fluoride (anti glycolytic agents) that
preserve glucose for up to five days. The tube is used
primarily to obtain glucose levels.
6. Free anticoagulants or additives (can be used for
blood typing and cross- matching).
7. Blood culture tubes (Yellow-black stopper)
During working in the laboratory:
Not allowed to eat in laboratories.
Tables should be kept clean and wiped by
sterilizing agents.
Wash hands in running water before and after
blood collection .
Gloves should be used in blood drawing and
handling.
Needle is disposed in sharp container
Needle stick injuries
Don't panic.
Clean the infected area.
Rinse with of water.
Wash with antibacterial soap
Squeeze wounded area
Place 70% alcohol on the wound for two minutes.
Contact the local health professional responsible for
infectious diseases.
1. Internal quality control
Each laboratory system should has quality
control materials.
In which, secondary calibrators should be applied
according to an internationally recognized
reference method. Each standard (calibrator)
should be run at least in duplicate
2. External quality control
External quality control is arranged by internationally
recognized reference laboratories that distribute batches of
samples of various concentrations for each assay.
In external quality control, The participating laboratory is
blinded to the concentration of the analyte. Standard
deviations of the results of the participating laboratory serve
as a measure of performance.
1. Individual analytical variation
2. Laboratory analytical variation
1. Individual analytical variation
a. Inter- individual variation
Age – Sex – Race – Genetics
b. Intra- individual variation
Diet – Drugs - Sleep pattern - Situation or Position
2. Laboratory analytical variation
a. Pre-analytical variation
Transport - Exposure to UV light
Standing time before separation of cells
Centrifugation time
Storage conditions
b. Analytical variation
Random errors
Systematic errors
c. Post-analytical variation
Transcriptions errors
Result reports to wrong patient
Hemolysis
Hemolysis ( Rupture of red blood cells) can cause significant
change in some analytics as potassium, magnesium, sodium,
……….
Red color of plasma or serum due to the hemolysis can affect the
results that are measured by colorimetric
Lipemia (lots of fats ) & proteinemia (lots of proteins)
Cause serum or plasma to be become turbid. This can affect
colorimetric and turbidimetric based tests ( give false results)
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Blood Samples - equipment of blood sample

  • 2. ➢Prof. Hend Hussein ➢ Ass. Dr. Samer Zahran Course Assistant: Ass. Lect. Sara El-Hadey
  • 3. Clinical Biochemistry course provide an advanced knowledge of human body chemistry levels under healthy and abnormal conditions. Also, it evaluates the suitability of clinical specimens and the laboratory test outcomes, and explain the relation between the diseases and the abnormality in blood chemistry.
  • 4. Clinical Biochemistry (PLC 402) involves the study of the proper sampling techniques, study of chemical components of the human body, plasma proteins, kidney, liver function, as well as the study of body electrolytes, acid-base balance, hormones and disorders in thyroid function.
  • 5. 1. Introduction 2. Blood Sampling 3. Acid-Base balance 4. Electrolytes 5. Kidney Function 6. Acute Phase Protein 7. Hormones 1 8. Hormones 2 9. Thyroid function 10. Diabetes Mellitus 11. Liver Function 12. Cardiac Function 13. Tumor Markers 1 14. Tumor Markers 2
  • 6.
  • 7.
  • 8. ➢ To diagnose diseases ➢ To screen for risk factors ➢ To monitor the effects of treatments & medications.
  • 9. In blood tests, blood sample is usually obtained from the patient's vein. Other specialized blood tests, such as the blood gases, require blood extracted from an artery. ➢ Blood gas analysis is used to monitor CO2, O2, blood pH and bicarbonate levels related to pulmonary function or certain metabolic conditions.
  • 11. Sodium, potassium, chloride, calcium, phosphate, magnesium, iron, urea, creatinine, liver enzymes, copper, zinc, glucose, cholesterol, bicarbonate, lactate, and others.
  • 12. ➢ Protein electrophoresis ➢ RNA ➢ DNA DNA testing (via PCR technique) is used in diagnoses of many disorders and forensic sciences.
  • 13. • Complete blood count (CBC) • Erythrocytes sedimentation rate (ESR) • Cross-matching (for blood transfusion or transplantation) • Blood culture
  • 14. Universal precautions should be used to increase the safety of blood collection. These precautions are important to reduce the risk of exposure skin and/or mucus membranes to infectious materials. It includes the use of a variety of protective barriers, such as gloves, masks, gowns (or white coat), and eyewear. Precautions
  • 15. Universal precautions were designed to prevent the transmission of HCV, HBV, HIV, and other blood pathogens. Universal precautions are applied in blood, semen, vaginal secretions, pleural fluids, cerebrospinal, synovial, peritoneal, pericardial, amniotic fluids and tissues. universal precautions
  • 16. You must avoid blood drawing from: ➢ arm affected by a stroke or neurological injury ➢ arm of a mastectomy ➢ areas with extensive scar tissues, shunt, varicose veins, and the palm ➢ arm of a patients that receive intravenous fluids. ➢ site that is swollen or affected by certain skin condition such as eczema or other infection
  • 17. Before the blood drawing, you should ask the patient about some factors that may affect test results: medications, pregnancy and age. Make sure the patient has followed special instructions such as fasting or taking a medication.
  • 18. The patient should be instructed about what blood tests have been ordered and why. Ask the patient if anytime has had trouble during the blood drawing. This question may be an alert to potential problems in finding an appropriate vein. Also ask the patients what position would be most comfortable for them when drawing the specimen. Venipuncture makes many patients nervous, and having them lie down.
  • 20. ➢ Needles: vacutainer needles, syringes, butterfly needles ➢ Antiseptic. 70% alcohol wipes or disinfection swabs ➢ Tubes ➢ Vacutainer holder ➢Tourniquet (is used to find a vein) ➢Adhesive dressing ➢ Rubber gloves ➢ Pillow or other support ➢ Sharps disposal container ➢ Disposal box
  • 21.
  • 22. The vacuum tubes are designed to draw a determined volume of blood. This system consists of vacuumed specimen tubes, a needle, and a plastic holder. When the tube is placed into the holder and pressed against the needle, negative pressure results, and blood is pulled into the tube
  • 23.
  • 24. ➢ Transfer and storage tubes ➢Pipettes ➢ Centrifuge ➢ Racks for tubes ➢ Special boxes for tube transfer and storage ➢ Freezer (for storage)
  • 25. 1. Venipuncture is useful to obtain cells and extracellular fluids (plasma or serum) from the body for analysis. 2. Fingerstick : If only a few drops of blood are needed (make a small prick in the tip of the finger and then squeeze the blood out). Ex: blood group
  • 26. 3. Heel-stick is the preferred method of blood collection on a newborn baby or infant
  • 27. During blood collection, the arm should rest on a pillow or other supportive prop. The patient open and close the fist. The phlebotomist sets the tourniquet around the upper arm, searches the proper vein by inspecting and palpating and then sterilizes the injection site. The use of a tourniquet should be minimized (to avoid changes in blood hemoconcentration).
  • 28. After that, the needle beveled upward, should be pushed smoothly and quickly into the vein, to minimize the possibility of hemolysis as a result of vascular damage. Immediately after the insertion, the tourniquet should be released (to minimize the effect of hemoconcentration).
  • 29.
  • 30. The needle will be attached either to a low pressure blood test bottle (Vacutainer), or to a syringe where the plunger is pulled back to create low pressure. When the necessary amount of blood has been extracted, the needle is removed and a little ball of cotton wool is held over the wound for 1 minute.
  • 31. The artery blood it is usually extracted from the wrist where the artery is very close to the skin. After taking blood from an artery, a ball of cotton wool is held over the wound about 5 minutes to stop any bleeding.
  • 32. Blood collection tubes come with a variety of colored stopper caps, and may contain additives. The colored stopper is important to avoid the cross- contamination of additives between tubes
  • 33. 1. Serum separator tube (SST). This tube contains a polymer gel for blood chemistry (Glucose, urea, cholesterol, Na, K, Mg Zn, ….. 2. Heparin tubes (green top tubes) are used for lithium and ammonia levels.
  • 34. 3. EDTA tubes ( with lavender top). For complete blood counts. 4. Sodium citrate tubes (light blue top) are used for prothrombin time (PT) and partial prothrombin time (PTT) analysis.
  • 35. 5. Sodium fluoride (anti glycolytic agents) that preserve glucose for up to five days. The tube is used primarily to obtain glucose levels. 6. Free anticoagulants or additives (can be used for blood typing and cross- matching).
  • 36. 7. Blood culture tubes (Yellow-black stopper)
  • 37. During working in the laboratory: Not allowed to eat in laboratories. Tables should be kept clean and wiped by sterilizing agents. Wash hands in running water before and after blood collection . Gloves should be used in blood drawing and handling. Needle is disposed in sharp container
  • 38. Needle stick injuries Don't panic. Clean the infected area. Rinse with of water. Wash with antibacterial soap Squeeze wounded area Place 70% alcohol on the wound for two minutes. Contact the local health professional responsible for infectious diseases.
  • 39. 1. Internal quality control Each laboratory system should has quality control materials. In which, secondary calibrators should be applied according to an internationally recognized reference method. Each standard (calibrator) should be run at least in duplicate
  • 40. 2. External quality control External quality control is arranged by internationally recognized reference laboratories that distribute batches of samples of various concentrations for each assay. In external quality control, The participating laboratory is blinded to the concentration of the analyte. Standard deviations of the results of the participating laboratory serve as a measure of performance.
  • 41. 1. Individual analytical variation 2. Laboratory analytical variation 1. Individual analytical variation a. Inter- individual variation Age – Sex – Race – Genetics b. Intra- individual variation Diet – Drugs - Sleep pattern - Situation or Position
  • 42. 2. Laboratory analytical variation a. Pre-analytical variation Transport - Exposure to UV light Standing time before separation of cells Centrifugation time Storage conditions b. Analytical variation Random errors Systematic errors c. Post-analytical variation Transcriptions errors Result reports to wrong patient
  • 43. Hemolysis Hemolysis ( Rupture of red blood cells) can cause significant change in some analytics as potassium, magnesium, sodium, ………. Red color of plasma or serum due to the hemolysis can affect the results that are measured by colorimetric Lipemia (lots of fats ) & proteinemia (lots of proteins) Cause serum or plasma to be become turbid. This can affect colorimetric and turbidimetric based tests ( give false results)