clinical pathology laboratory activity ems 2012 blood glucose measurement kketone and glucose urine...
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Clinical Pathology Laboratory Activity EMS 2012
• Blood Glucose Measurement• Kketone and Glucose Urine Exam
• Blood Gas Analysis• Cholesterol Examination• Cortisol Measurement
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Factors influencing laboratory results
Pre analytic
Analytic
Post analytic
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Glucose Measurements
SPECIMEN CONSIDERATIONS
Whole blood Plasma Serum Pleural fluid CSF Urine
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Most measurements by enzymatic methods:
1. glucose dehydrogenase, 2. glucose oxidase, 3. hexokinase
Glucose Measurements
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1. Glucose Dehydrogenase
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2. Glucose Oxidase,
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3. Hexokinase
(Henry JB. Clinical Diagnosis and Management by Laboratory Methods. 2011)
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• These reactions produce an electrical current that is proportional to the initial glucose concentration, or a product that measured spectrophotometrically is proportional to the initial glucose concentration.
• The assays can be initial rate-of-change assays, where the velocity of the reaction is dependent on the initial glucose, or end-point assays.
Glucose Measurements
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Spectrophotometer
POCT (point of care testing)/ Blood Glucose Home Monitoring/ Self Glucose Home Monitoring
Glucosemeter
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Spectrophotometer
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POCT (point of care testing)/ Blood Glucose Home Monitoring/
Self Glucose Home Monitoring
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Method of POCT (point of care testing)/ Blood Glucose Home Monitoring/
Self Glucose Home Monitoring
Biosensor
Reflektansmeter
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Biosensor Method
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Biosensor Method
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Reflektansmeter/ Amperometer Method
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A wide variety of devices are available forhome measurements:
• avoiding operator errors• Calibration• Errors that may contribute to inaccurate readings in certain
devices include the application of :- an insufficient volume of blood- milking the finger to acquire sufficient blood- the use of outdated test strip- environmental factors (humidity, heat, altitude)- the use of a malfunctioning meter- the use of a dirty meter - hypertriglyceridemia- hypotension- measurements outside of the hematocrit or temperature range
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Influecing Factors
• Influenced by high levels of salicylate, acetaminophen, levodopa, uric acid, bilirubin,lipids, or low oxygen levels, and others are altered by touching the reaction area.
• Vit C, lactose, manose, galactose, xylose .
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Reference Value
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Ketone and Glucose Urine Exam
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Dipstick Urine Test/ Strip
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Urine test strip:Rapid, easy, specific,cheap
SPECIFIC GRAVITY
PROTEIN
KETOBODY
UROBILINOGEN
BLOOD
PLASTIK ROD
NYLON COVER
NITRITE
GLUCOSE
TEST FIELD(PAPER CONTAIN REAGENT)
FILTER PAPER
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DIPSTICK TEST
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Dipstick urine test
1. Specific gravity2. pH3. Leukocytes4. Nitrates5. Protein6. Glucose7. Ketones8. Urobilinogen9. Bilirubin10. Blood
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Glucose (negative)
Glucose is normally present in glomerular filtrate, but it is reabsorbed by the proximal tubule.
D-Glucose + O2 δ-D gluconolactone + H2O2
GOD
H2O2 + 0-tolidine H2O + colourPOD
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Ketones (negative)
in healthy individuals, ketone bodies are formed in the liver and are completely metabolized so that only negligible amounts appear in the urine
sodium nitroprusside and glycerin + acetoacetate and acetone
alkaline medium violet dye complex.
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URINALYSIS STEPS
PRE-ANALYTIC : patient preparation, samples collection, samples handling, labelling, refrigeration, preservatives of urine specimens.
ANALYTIC: principle of procedures, measurements, interpretation, conventional & rapid and sophisticated
POST-ANALYTIC: recording, reporting, use of units : conventional unit and international unit
QUALITY CONTROL (QC):calibration, control solution to get good and reliable results
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Pre-Analytic1. Specimen collection- Requisition form must accompany with specimen
delivered to the lab; the form include : patient’s name, I.D number, date and time of collection and additional information : age, location, physician’s name, type of specimen/method, interfering medication and clinical information
- Container clean, dry, leak proof, disposable - All specimens must be properly labeled must be attach
to the container, not to the lid, should not become detached if the container is refrigerated
- The information on form requisition match with the inform on the label
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Labelling urine specimen
Name (min 2 initial, ex: Deni Darmawaty)
Sex : female
Date (day, month, year)
Time collection:
Adress and telp number :
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Pre-AnalyticTwo Type of Specimen Based on Time :
- Random specimen- First morning/fasting specimen- 2-hours post prandial specimen- Timed urine (12-hours, 24-hours specimen)
Based on method :- Midstream Clean-Catch specimen- Catheterized specimen- Suprapubic aspiration
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Type of Urine Specimens Purpose
Random Routine screening
First morning Routine screeningPregnancy testsOrthostatic protein
Fasting (second morning) Diabetic screening/monitoring
2-h postprandial Diabetic monitoring
Glucose tolerance test Accompaniment to blood samples in glucose tolerance test
24-h (or timed) Quantitative chemical tests
Catheterized Bacterial culture
Midstream Clean-catch Routine screeningBacterial culture
Suprapubic aspiration Bladder urine for bacterial cultureCytology
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• The first morning urine:
Collected upon rising, it represents the urine over
approximately an 8 hour period
• Ad random urine:
Collected any time
• The 2-hour postprandial urine:
Collected 2 hour following the meal ( for urine glucose)
• The 24-hour urine:
A pooling of all urine excreted by the patient over a 24 hour period
(for protein, uric acid, calsium quantitation, etc)
• Midstream urine:
The middle portion of a single urination
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Pre-Analytic
3. Specimen HandlingFolowing collection should be delivered to the lab promptly and tested within 1-2 hoursIf it can’t delivered must be refrigerated or add with chemical preservative
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LIMITATION OF THE METHOD AND INTERFERING FACTORS:
• Highly pigmented or large amounts of levodopa metabolites in urine may cause weak positive result
• High Specific Gravity and low pH urine and PSP (phenolsulfophthalein) may cause false positive result
• High-protein, carbohydrate-free, high-fat diets may result in ketonuria (false posi- tive)
• Medications : phenazopyridine , ascorbic acid, ether, insulin, isopropyl alcohol, metformin, isoniazide, isopropanol, paraldehyde, valproic acid, and bromsulfoph- thalein also can cause false positive results
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EXPECTED VALUE
• In starvation diets or in other instances of abnormal carbohydrate metabolism, ketone appear in urine in excessively large amounts before serum ketone are elevated.
• This test detected 5 mg/dL of aceto-acetic acid; 70 mg/dL acetone; but more specific for aceto-acetic acid.
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Lipoprotein structure