Automated urine analysis.pptx final.pptx

Urinalysis

1. AUTOMATED URINE ANALYSIS- ADVANTAGES AND PITFALLS PRESENTED BY: DR.SHAILESH TADHA(FIRST YEAR RESIDENT ) MODERATED BY: DR.PURVI PATEL(ASSOCIATE PROFESSOR) GUIDED BY: DR.VIRAL BHANVADIA(ASSISTANT PROFESSOR)

2. TOPICS TO BE COVERED 1. Why automation? 2. Automated urinalysis features. 3. Options in automation. 4. Automated urine chemistry analysers- principles. 5. Automated urine microscopy analysers -principles. 6. Automated urinalysis systems. 7. Calibration & Quality Control. 8. Comparison between manual & automation. 9. Pitfalls in automation. 10 . Summary.

3. WHY AUTOMATION? Increases productivity. Labour savings. Improvement in turn around time. Performed in uncentrifuged urine. Reduces errors and standardise sample processing, biochemical strip analysis , microscopic analysis & report results. Better response to clinician’s and administration’s concerns. Improves precision and accuracy of data. Ease of use.

4. AUTOMATED URINALYSIS FEATURES 1.On-line computer availability. 2.Bar coding. 3.Sorting of patients and control results. 4.Calibration. 5 . Cleaning & maintenance.

5. OPTIONS IN AUTOMATION… 1.Automated urine chemistry analysers 1. Semi-automated analyser - The test strip is dipped in the urine manually and analysed by an instrument. 2. Fully automated analyser - The test strip is analysed completely by an instrument. 2.Automated urine microscopy analysers 3.Automated Urinalysis systems

6. DIPSTICKS Microchemical systems permit qualitative and semiquantitative analysis within a minimum time span by simple observation. Reagent impregnated paper & Absorbent paper underneath are held in place on a stiff white carrier foil by white Nylon mesh. Different reagent areas are affixed on the strip.

7. PARAMETERS: Bilirubin Urobilinogen Ketones Ascorbic acid Glucose Protein Blood pH Nitrite Leucocyte Compensation field Specific gravity

8. DIFFERENT DIPSTICKS Uristix Glucose, Protein Multistix-SG PH, Specific gravity, Glucose, Protein, Ketone, Bilirubin, Blood, Urobilinogen Combur 10 M, LabStrip U11PLUS Also Nitrite & Leucocyte Ascorbic acid Combistix-SG PH, Specific gravity, Glucose, Protein Keto-DIASTIX Ketone, Glucose

9. ADVANTAGES OF DIPSTICK AUTOMATION Enhances work flow by saving labour and time. Reduces subjective errors/Observer bais. Standardizes some aspects of manual urinalysis. Large number of samples in short time.

10. 1.URINE CHEMISTRY INSTRUMENTS AND PRINCIPLES Semi-automated Chemistry Instruments: Require operator to activate instrument, measure and enter the color and clarity of specimen. Manually dip and blot the reagent strip and place the strip on platform in the strip reader. Instrument detect and read reflectance of each reaction pad at proper time. F inally results are displayed on the screen.

11. • Semi-automated Chemistry Instruments EQUIPMENT MANUFACTURER Clinitek 500 Clinitek Status+ Siemens Healthcare Diagnostics Clinitek Advantus UC-1000 Sysmex Corporation Urisys, cobas u411 Roche Diagnostics

12. cobas u 411 Semi automated instrument Works on reflectance photometry Useful in lab. with workload of 30-100 sample per day Only compatible with Combur 10 M strip .

13. Fully-automated Chemistry Instruments Multiple barcoded tubes of urine are loaded into a rack. Sample identification by barcode reading and automatic pipetting of a precisely measured urine aliquot directly onto a reagent strip. Reagent strip is advanced to a reflectance photometer that measures the color change of each reaction pad at the proper time and at the appropriate wavelength. Abnormal results are flagged to identify specimens requiring microscopic examination or further testing.

14. • Fully-automated Chemistry Instruments EQUIPMENT MANUFACTURER Clinitek Novus Siemens Healthcare Diagnostics UC-3500 Sysmex Corporation LabUMed 2 77 Elektronika Cobas u 601 Roche Diagnostics

15. CLINITEK Novus Fully Automated Works on reflectance photometry principle Dry-pad urine chemistry technology 240 smples/hr

16. URINE CHEMISTRY INSTRUMENTS - PRINCIPLES REFLECTANCE PHOTOMETRY – Clinitek Status+, Urisys 1100,Cobas u 411,Chemistrip Super automated Analyser. ELECTROCHEMILUMINISCENCE – Cobas 6000 e601

17. REFLECTANCE PHOTOMETRY: Narrow beam of monochromatic light(from LED) is focussed on the dipstick pad. Darker the test pad, less light is reflected. Amount of light reflected from the chemical reaction is measured.

18. The reflected light is detected by a photodetector , which is converted by analogue digital coverter (ADC) and processed by microprocessor creating a signal that can be quantitated.

19. UriPlus 900 Fully automatic Works on Reflectance photometery 14 parameter 240 sample/hr

20. ELECTROCHEMILUMINISCENCE(ECL) Light emitting substance ( Ruthenium labelled Ab + Tripropylamine ) is added to the sample. Ruthenium & TPA are non isoptopic & highly stable at base state. Excitation & Emission of light (photon) is electrically stimulated. The peak of light can be continuously reproduced through cyclic amplification of the signal and accurately detected by a photo- multiplier. Amount of light produced is directly proportional to the amount of substance to be detected.

21. Cobas 6000 e601 Fully automated Works on ELECTROCHEMILUMINISCENCE 100 sample/hr

22. 2.AUTOMATED URINE MICROSCOPY ANALYSERS- PRINCIPLES Principle Machine Flow cytometry UF-1000i,UF-5000 analyser,AUTION HYBRID Digital flow microscopy Iris iQ200 microscopic analyser,URISED Cuvette-based digital microscopy 77 Elektronika- URISED 3 PRO

23. FLOW CYTOMETRY(FCM) FCM is a LASER based technology which identify & quantify cell populations. The analyser passes the sample as a LAMINAR FLOW through a LASER BEAM. 2 dyes are used : 1. Phenathridine (orange) - DNA 2. Carbocyanin (green) - cell membranes, nuclear membranes, mitochondria

24. 5 components: 1. LASER - provides monochromatic light. 2. FLOW CELL - with a liquid stream ( sheath fluid ), which carries & aligns the cells so that they pass in single file through the laser beam. 3. OPTICAL SYSTEM & filters regulating the light signals. 4. PHOTOMULTIPLIER DETECTORS (PMTs) that generate data on forward light scatter (provides approximation of cell size ) & side light scatter (relates to cell complexity ), as well as converting fluorescence signals from light into electric signals that are processed by a computer. 5. COMPUTER for analysis of signals.

25. FCM relies on the application of fluoroscent conjugates or FLUOROCHROMES attached to a monoclonal Ab that has specific affinity for an Ag in the cell. Each fluorochrome has a characteristic peak excitation & emission wavelength . RESULT- obtained in form of Scattergrams or Histograms.

26. UF-50,100,1000i give quantitative result for RBC, WBC, Epithelial cells, casts, bacteria yeasts, crystals, dysmorphic RBCs. Printout from UF100-NORMAL URINE : absence of RBCs, WBCs,bacteria & casts

27. ABNORMAL URINE- SIZABLE NO. OF RBCs, WBCs, Bacteria & casts

28. SYSMEX UF-1000i Works on Flow cytometry. Two stains with fluorescent dye Parameters: RBC,WBC, Epithelial cells,Casts, Bacteria. 100 sample/hr

29. AUTO PARTICLE RECOGNITION / DIGITAL FLOW MICROSCOPY The sample is mixed & aspirated to a planar flowcell where 500 digital photomicroscopic images are taken per sample . The system uses Auto Particle Recognition software that classifies urine particles into 12 categories based on size, shape, texture & contrast into – RBCs, WBCs, WBC clumps, squamous epithelial cells, non squamous epithelial cells, hyaline casts, unclassified casts, bacteria, yeast, crystals, mucus & sperms.

30. IRIS IQ 200 Automated urine microscopy analyser. Works on Digital flow microscopy principle Coupled with Auto Particle Recognition (APR) Software

31. IRIS FLOW VIDEOMICROSCOPY Same as a digital flow microscopy but instead of using images machine take the video snaps. Video snaps are sorted by the computer. Computer analysed the all information and final results are produced. Examples of pictures/snaps ↓

32. CUVETTE-BASED PARTICLE ANALYSIS Recent innovation. Urine specimen centrifused in a cuvatte. Multiple images captured by high resolution digital camera coupled to a microscope. URISED 3 PRO , UriSed/Sedimax and Cobas U 701 are some examples.

33. UriSed 3 PRO Works on Cuvette based automated microscopy and image processing. Coupled with Phase contrast,Bright -field and composite images. Parameters: Almost All. 150 Sample/hr.

34. 3.AUTOMATED URINALYSIS SYSTEMS Provide integrated analysis of urine chemistry and microscopy. Chemical analysis is performed first, and the specimen is automatically transported to the microscopy unit. Optimal for larger , high-volume laboratories. iRICELL Automated Urinalysis System : iChem Velocity urine chemistry + iQ200 microscopy analyzer. CLINITEK AUWi System : ATLAS + UF-1000i flow cytometer. LabUMat chemistry analyzer + UriSed microscopy unit . AUTION HYBRID System : Only completely integrated urinalysis system as single compact unit.

35. iRICELL Automated Urinalysis System Combines urine chemistry and microscopy. Works on Auto-particle Recognition principles. Deliver standardized and accurate results.

36. CALIBRATION & QUALITY CONTROL Calibration should be done every 6 months ( Clinical Laboratory Improvement Amendments (CLIA) regulation) or as according to instrument specification. Fully automated instruments – calibrations & quality controls are internally regulated & done according to instrument specification . Semi-automated chemistry instruments utilise calibration strips for this purpose. 1. Calibration is required before using the instrument, whenever we change LOT NO. of the strips, or as prompted by the instrument. 2. Check the expiry date of the calibration strip before using it 3. Close the container immediately after removing the strips.

37. In our lab we do Quality Control manually once every morning, using level 1 & level 2 & results recorded according to pre-set ref. ranges REAGENT – Prepared from human urine with added human RBCs, simulated leucocytes, constituents of animal origin & preservatives. After calibration, Quality Control can be performed using LEVEL1(Normal control) & LEVEL2(Positive control) on the test strips .

38. PITFALLS IN AUTOMATED URINALYSIS Expensive maintenance budget. Needs technical training for instrument operation. Crash of automat( need backup )- a good support & maintenance required. Knowledge and understanding of the interference factors is required for correct interpretation of the results. Flagged results in automated cell analysers require manual microscopy for confirmation. Iris IQ200 mistakes RBCs for calcium oxalate crystals(ovoid type), requiring manual confirmation.

39. SUMMARY So a complete urinalysis using macroscopic examination, reagent strips, automation and manual microscopy can be used in conjunction for standard and accurate results. Automation has more benefit in setups with high sample load . Automation helps to screen samples for UTI or kidney disease.

40. THANK YOU