Pars Azmon (Automation) Pars Azmon (Manual) Biosystem (Automation) Biosystem (Manual) Ziest Chemi (Manual) 62
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1 STANDARDIZATION OF URNIE PROTEIN MEASURMENT R. Mohammadi Biochemist (Ph.D.) Faculty member of Medical Faculty
2 EQAP 7 : URINE PROTEIN Method No. Mean (mg/dl) CV% Pars Azmon (Automation) Pars Azmon (Manual) Biosystem (Automation) Biosystem (Manual) Ziest Chemi (Manual) Bahar Afshan (Manual)
3 EQAP 8 : URINE PROTEIN Method No. Mean (mg/dl) CV% Pars Azmon (Automation) Pars Azmon (Manual) Biosystem (Automation) Biosystem (Manual) Ziest Chemi (Manual) Bahar Afshan (Manual)
4 EQAP 9 : URINE PROTEIN Method No. Mean (mg/dl) CV% Pars Azmon (Automation) Pars Azmon (Manual) Biosystem (Automation) Biosystem (Manual) Ziest Chemi (Manual) Bahar Afshan (Manual)
5 CV% OF URINE PROTEIN MEASURMENT Method EQAP 7 EQAP 8 EQAP 9 Pars Azmon (Automation) Pars Azmon (Manual) Biosystem (Automation) Biosystem (Manual) Ziest Chemi (Manual) Bahar Afshan (Manual)
6 ON THE BASIS OF CAP PT SURVEYS Methods used to quantitate urinary protein have not been satisfactory. Mean values reported vary twofold between methods, with SSA method producing high values. Precision is poor, with SSA showing the poorest CV. TCA-biuret, Coomassie blue, and TCA show closer agreement and about half the CV of SSA. Problems arise from nonstandardized methods. With turbidity methods these include different and concentration and timing, and variation in protein used.
7 URINE PROTEIN In Normal State, Maximally There Is 150 mg/24h or 10 mg/dl Protein in Urine More Than 200 Protein Have Been Found in Urine Including 1/3 Albumin 1/3 Tomm-Horsfall 1/3 Other Proteins
8 URINE PROTEIN In Normal State, Maximally There Is 150 mg/24h or 10 mg/dl Protein in Urine More Than 200 Protein Have Been Found in Urine Including 1/3 Albumin 1/3 Tomm-Horsfall 1/3 Other Proteins
9 TYPES OF PROTEINURIA Classification Based On Origin Prerenal Dehydration, Decreased Blood Flow, Increased Pressure on Kidney, Fever, Infectious Disease Renal Glumerolar, Tubular Postrenal Inflammation, Infection, Stone
10 TYPES OF PROTEINURIA Classification Based On Amount of Secreted Protein Microproteinuria Minimal Proteinuria Moderate Proteinuria Severe Proteinuria
11 TYPES OF PROTEINURIA Classification Based On Time of Proteinuria Transient proteinuria Intermittent Proteinuria Orthostatic Proteinuria Constant Proteinuria
12 LABORATORY INVESTIGATION OF PROTEINURIA Screening Strip Test Confirmatory Test Quantitative Total Protein Measurement Specific Protein Detection or Determination
13 SCREENING STRIP TEST Based on Error of Indicator It Is Sensitive for Albumin, But Not Globulins Interference with Bilirubin and Drugs Such as Phenoazopyridine Which Give Orange Colour False Positive Results 1) Urine alkaline ph 2) Increased contact time with urine 3) Contamination with quaternary ammonium compounds or chlorohexidine False Negative Results 1) Increased Salts 2) Protein Other than Albumin
14 CONFIRMATORY TESTS Based on Denaturation of Proteins Detects Albumin & Also Globulins 1) With SSA, turbidity for albumin is 2.5 times of globulins 2) with TCA, turbidity for globulin is greater than albumin, but this difference is not marked False Positive Results 1) Turbid urine 2) Radiographic contrast media 3) Drugs such as penicillin, sulfonamides or Cephalosporin False Negative Results Highly alkaline urine ph
15
16 BENCE JONES PROTEIN (BJP) Strip Is Neg, But SSA Is Pos The Best Method Is Electrophoresis Heat Test Has A Low Sensitivity False Results with Heat Test Are Due to : 1) Low concentration 2) High concentration 3) Presence of other proteins 4) All BJP do not react with this test
17 STANDARDIZATION OF URINE PROTEIN QUANTITATION Urine Collection Method of Analysis Calibration Calculations and Reporting
18 URINE COLLECTION Timed Urine Specimens Preservatives Patient Activity
19 METHOD OF ANALYSIS Biuret Method Precipitation Methods, such as SSA and TCA Dye-binding Methods, such as Pyrogallol red or CBB
20 CALIBRATION OF METHOD Bovine or human albumin Diluted Serum (or serum pool) with a normal albumin/globulin ratio, and analyzed for total t protein with a correctly calibrated and well controlled biuret or Kjeldahl s method
21 CALCULATION AND REPORTING Unite of urine protein analyze may be mg/dl or mg/l Unite of urine protein reporting may be mg/dl, mg/l, mg/12, or mg/24h
22
URINE PROTEIN In Normal State, Maximally There Is 150 mg/24h or 10 mg/dl Protein in Urine More Than 200 Protein Have Been Found in Urine Including 1/3
URNIE PROTEIN R. Mohammadi Biochemist (Ph.D.) Faculty member of Medical Faculty URINE PROTEIN In Normal State, Maximally There Is 150 mg/24h or 10 mg/dl Protein in Urine More Than 200 Protein Have Been
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