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Clinical laboratory
The document provides an overview of the clinical laboratory, including its roles, workflow, common specimen types, and sampling/testing procedures. The key responsibilities of the clinical lab are the correct identification, collection, and processing of patient specimens; accurate testing; timely reporting of results; and communication with healthcare professionals. There are typically six main steps in how a sample flows through the lab: ordering a test, sample collection, delivery to the lab, processing, analysis, and reporting results. Common specimen types include blood, urine, body fluids, sputum, stool, and tissue samples.
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Clinical laboratory
- 1. CLINICAL LABORATORY BASIC OVERVIEW SuryaniAdawiyah Application Specialist
- 2. Laboratory Roles andResponsibility The responsibilities of the clinical lab include : Correct Identification, collection and processing of patient speciments Accurate performance of testing Timely reporting of result Communication with physicians and other healthcare proffessionals Analyst testing is used to help diagnose, monitor and treat disease
- 3. Laboratory Workflow There are6 main steps in how a sample flows through the lab from other creation to final test result 1. Test ordered 2. Sample is collected 3. Sample is delivered to the lab 4. Sample is processed 5. Sample is analysed 6. Result are reported Test Order Sample collected Result report Sample Deliver to lab Sample analysed Sample Processe d
- 4. Laboratory Speciment Most commonlaboratory specimens types : Blood Urine Additional Laboratory specimens : Body Fluid Sputum Stool Tissue samples Culture swabs
- 5. Sampling / Phlebotomy Typesof Samples : Serum, Plasma, Blood, Urine, CSF
- 6. Why do weanalyze our blood Corronary disease : CK, LDH, Troponin T
- 7. Why do weanalyze our blood Kidney Disease BUN, Creatinine Liver Disease Bilirubin
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- 9. Medical Examination Objective isto get an answer about the health status of patient The physician determines on the basis of the anamneses, his clinical examination and on the basis of additional known information an enquiry → Examination Request His is Followed by the necessary preparation of the patien and blood sampling
- 10. What is Bloodcomposition ? Blood Composition : • Plasma • Cells 55 % Plasma • Yellow, Sticky liquid • Transport of nutrients ( protein, Fats, Carbon Hydrates ) • Hormones 44 % Erytrocyte • Red Blood cells • Contain haemoglobin • O2 & CO2 transport
- 11. Sample Container Whatdo we use ? Different type of sample collection in comercially available blood collection system (Beckton Dickinson Vacutainer, Sarstedt Monovetten,…) Anti Coagulant agent Ext Application Colour Non coagulant Serum Clin chem, sero, immun Red heparin plasma Clin chem Green EDTA Plasma Hema, special chem, immun Lilac Citrate Plasma Coagulation test PT/APTT Bleu Na-Fluoride/K-Oxalat Plasma Glucose, Lactat Grey Pink Yellow
- 12. What samples dowe analyze ?
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- 14. Pre-Analytical Possible Influences Age gender Genetic Influences Nutritional Influences Pregnancy Biorhythm (diurnal rhytm causing analytical fluctuations) Muscullar mass, body weight Physical activity or inactivity Psychological stress ( fear for blood collection, surgery) Use of medicines
- 15. Pre-Analytical Disturbing Influences Samplecollection (body position, venous congestion, …) Sample condition (haemolytic, Lipemic, Icteric) Normal serum obtained from an individual in good health is usually clear, pale yellow in collor. However, the color of the patients serum may appear different for various reasons such as disease or improper handling of the blood specimen. Lipemia (lipe) result from increased levels of lipoproteins associated with triglycerides, and it can cause the serum to appear white. Hemolysis (heme) is caused usually by the release of hemoglobin from ruptured red blood cells during sample collection and/or sample handling this inteference can cause the serum to appear red
- 16. Pre-Analytical Icterus (Icte) isthe result of increasing levels of bilirubin, and it can cause the serum to appear yellow.
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- 18. Pre-Analytical After the centrifugationif the sample was without anticoagulant The supernatan fluid is SERUM otherwise is plasma. As anticoagulants they use EDTA K3, EDTA K2, Heparin, Citric Acid (:1, Citric Acid 4:1 NaF and others. If we use plasma we must know the type of the anty-coagulant due to different interferences Ca, Na, Fe, ALP
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- 20. Pre-Analytical Some photometricassays may be influenced by the presence of these abnormal serum colors and the reliability of the test result may be decreased. Haemolysis can cause analytical interferences such as high K+ caused by release from erythrocytes, or can interfere with the measuring technique (photometry) Inadequate sample transport Wrong centrifugation Inadequate sample storage (bilirubin)
- 21. Pre-Analytical Serum • 30-45 Minutesclothing (preferably in the dark) • 10-15 minutes centrifugation @ 1000-1500 9 Plasma • Immediate 10-15 minutes centrifugation @ 1000-1500 g
- 22. Pre-Analytical Sample transport andstorage Properly packed Transport must be save biohazardous material 4 hours stable @ 15-25⁰C closed to avoid evaporation 24 hours stable @ 4-8⁰C Dry Ice, cool packs, refrigerator, etc
- 23. Pre-Analytical Example : Potasium Plasma is recommended for rapid centrifugation, use only serum or plasma for single patients Sample preparation (heparin plasma), centrifuge within 30-45 minutes after collection, erythrocytes produce hemocysteine, which continues after sampling Store on ice if centrifugation within 30-45 minutes is not possible Store plasma at 20⁰C if sample can be measured within 48 hours
- 24. Analytical Adequate test methodology Standardoperating procedure Understandable Traceable Routine test must be Easy to be executed Reliable Low risk failure
- 25. Statistical Quality Control Sampleswith known concentration Low Medium High As part of daily routine Begin of the run Middle in the Run End of the day Random
- 26. Pre-Analytical Test Report Demographic Information Patient name, Patient ID, Lab number Sample matrix, visual distortions Date, Time sample collection, arrival in the lab, time of analyses Analytical results Test name, Unit Reference values, comments ( High/Low, diluted, duplicates,…)
- 27. Analytical Result Expected Values Referencerange Normal Values Based on a large pool of healthy persons Differences between Children vs adult Male vs female Serum vs plasma Population biorhytem
- 28. Diagnose After checking thereliability of the analysis Analytical range Statistical Quality Control Pre-analytical and analytical disturbances Plausibility of the result Compared with previous result Fit with the situation of the patient
- 29. Method Of ClinicalChemistry Photometry chemiluminence Potentiometry (ISE) Electrophoresis Nephelometry Y-Counter Mass Absorption Osmometry
- 30. Photometry In photometry, analiquote of sample containing analyte is mixed in a cuvette with a liquid reagent. The reagent react with analyte producing a change in absorbance (color) within the reaction solution. The absorbance is measured using a photometry system
- 31. Photometry This is achieveby comparing the amount of transmitted (Is) light to the amount of light entering (I0) The change in absorbance is proportional to the concentration of analyte in the sample. Typically, more analyte in the sample generates a darker colored solution in the cuvette, thus, less light gets through to the detector.
- 32. Linear Calibration Curve (LinearReaction) 600 500 400 300 200 100 0 0 100 200 300 400 Slope = angle of line 500
- 33. Calculation If a blankand only one calibrator are run, the factor is determined as : F Conc std Concstd = Rstd - Rblk Where : Conc std = concentration of the calibrator R std = response of the calibrator Rbllk = response of the blank Type of Calibration Number and type of Calibration Conversion into concentration Absolute calculation ( ABS calc) Reagent Blank Reaction ABS X FV One point calibration curve (STD calc) Reagent blank one standard Reaction ABS X FV/(STDABS)
- 34. Rate Method Rate reaction( Reaction change as a function of time): using this principle, a result is calculated from the change in signal per unit of time. The rate of the signal change is measured. These reactions can also be described as either up and down. Enzymes are measured using the rate reaction. Examples of rateup are CK and LDH. Examples of rate ALT and AST. Rate-Up Reaction
- 35. Rate or ZeroOrder Kinetics RRA is the rate method of obtaining concentration or activation value from absorbance change per minute between two points using the least – squares method L, m, n p, r Measurement points S sample volume V, Vp, m Reagent Volume Amn Mean Absorbance, exclude min&max Δamn The change in absorbance per minute between measurement Tmn Time (min) between measurement points m & n A(tp) Absorbance obtained by substituting the time at measurement point into the approximation curve ΔA(tp) The slope of the reagent to the approximation curve ( the change of absorbance per minute) Kpm Liquid – volume correction coeficient Example of setting general reaction process (decrease reaction) and measurement points using the RRA method Kpm = (S+Vp)/(S+Vm)
- 36. Rate, Zero Order Decrease: 340 nm AST/GOT-ALT/GPT, LDH P—L, Aldolase FACTOR or FV = (Vtotal X 1000) / (Vsample X Light Path X MEC ) Increase : 340 nm : LDH L- P, CK, CKMB, HBDH, ELASTASE, LAP 405 nm : ALP, ACP, NP Factor or FV = ((Vtotal X 1000) / (Vsample X Light Path X MEC )
- 37. Turbidimetric Assay Turbidimetric Assay Turbidimetricassays measure the intensity of the transmitted light as shown below. Early turbidimetric assay Were Not sensitive enough To Measure low levels of Serum proteins. However, significant improvements in newer automated analyzer have made Turbidimetric assays equivalent to nephelometric analysis Turbidimetry Principle Based on the principle of measuring the intensity of transmitted light. A. Incoming Light B. Transmitted Light
- 38. Potentiometry Potentiometry is basedon electronical reaction and is the measurement of the electrical potential between two electrodes in an electronial cell. Examples of analytes that typically utilize potometry for their measurement are the electrolytes sodium (Na+), potassium (K+) and chloride (Cl-). Ion selective membrane electrodes ( ISE) are utilized with spesific permeability to selected anions and cations (e.g, Valinomycin membrane to measure (K+) . Sample containing analyte is brought into contact with the ion specific membrane. Concentration are calculated from the measured potential through the Nernst equation.
- 39. Potentiometry Direct potentiometry :this is the simples method of making ionselective electrode measurements. The electrodes are immersed in test solution and the electrode potential is measured directly to this measurement by reading the answer from a calibration graph of concentration versus millivolts. Indirect potentiometry : dilution of the sample (less volume, less problems, less interventions)
- 40. Case Example Proteins arepresent in all body fluids. Their concentration is normally high only in blood, serum, plasma, lymph fluid, and some exudates. There is a small amount of proteini in spinal fluid and trace of protein in urine. Where do you find high levels of proteins Proteiin have many purposes. They function as antibodies, form part of the endocrine system, and provide a complex blood-clotting system. Additionaly, they are carriers for other compounds, provide tissue nutrients, and function as enzymes. To determine disease processes it is important to compare levels for each fraction of the proteins to normal values
- 41. Pre-analytical factors thataffect serum proteins consentration 1) 2) 3) 4) 5) 6) 7) 8) 9) 10) Time of the day Position Exercise Fasting vs non fasting Medications Time of year (season) Age and gender Geographic location Venipuncture technique Sample handling and storage
- 42. Patient Result Test Report DemographicInformation • Patient name, Patient ID, Lab Number • Sample Mtrix, Visual distortions • Date, time sample collection, arrival in the lab, time of analyses Ana
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Editor's Notes
- #3 The clinical lab provides diagnostic test data to aid in the detection, diagnosis and treatment disease. Data is used by physicians, nurses, pharmacist and other healthcare professionals