VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE Clinitek® 50 Urine Chemistry Analyzer is used for glucose, bilirubin, ketone, specific gravity, occult blood, pH, protein, urobilinogen, nitrite, and leukocytes testing. The Point of Care Testing (POCT) program at Vanderbilt University Medical Center (VUMC) is overseen and coordinated by the POCT Steering Committee as chartered by the Medical Center Medical board. The POCT Steering Committee administers the program with the primary goal of meeting clinical needs in the most cost-efficient manner possible. In order to provide for both regulatory compliance and appropriate utilization of POCT, any clinical area wishing to perform POCT must be approved by the POCT Steering Committee and must maintain acceptable performance according to criteria set forth in Hospital Policy 20-23. I. PRINCIPLE: A. Instrument- Reagent test strips saturated with urine are blotted and placed on the Clinitek 50 table. The table is pulled inside the instrument where each test pad is measured for color and amount of light reflected. An incandescent lamp inside the instrument illuminates a fixed “read area” on the reagent test strip. Light reflected from the test strip surface is detected by a photodiode pack, containing four (Red, Blue, Green, and IR) filters. The reflected light is then converted into electrical impulses, which are processed by the instrument’s microprocessor and converted into clinically meaningful results shown on the display screen or printed by the internal printer. B. Multistix 10 SG test strips- The reaction of Bayer Multistix 10 SG test strips depends on color development as an indicator of the concentration of the following test reactions. The pH test reaction uses methyl red/bromthymol blue indicator system, which provides pH readings from 5.0 - 8.5. The color of the test pad changes from orange to green to blue in this range of pH. The glucose test is based on a double sequential enzyme reaction, which requires glucose oxidase, peroxidase, and potassium iodide. A glucose specific enzyme, glucose oxidase, catalyzes the formation of gluconic acid and H202 from the oxidation of glucose. The enzyme peroxidase catalyzes the reaction of H202 with a potassium iodide chromagen to oxidize the chromagen to colors ranging from green to brown. Page 1 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE The protein reaction uses tetrabromophenol blue reagent buffered at a pH of 3. This colorimetric method is based on a concept known as the “protein error of indicators,” where the point of color change of certain buffered pH indicators is different in the presence or absence of protein. Colors range from yellow (negative) through yellow-green, green, and blue-green, as positivity increases. The protein test pad is more sensitive to albumin than globulins (such as gamma-globulin, Bence Jones, hemoglobin, and mucoprotein), and a negative result does not necessarily rule out the presence of these other proteins The ketone reaction uses sodium nitroprusside reagent, which, in the presence of acetoacetic acid produces a color change from buff-pink (negative) to purple as ketone concentration increases. Acetone and B-hydroxybutyric acid are not detected with this method. The bilirubin reaction occurs when bilirubin reacts with diazonium salt (2,4 dichloroaniline) to form a tan color. Urobilinogen reaction is due to the reaction of urobilinogen with pdiethylaminobenzaldehyde, (in an acid medium), producing a pink-red color. Leukocyte Esterase reaction is due to derivatized pyrrole amino acid ester and diazonium salt. Leukocytes of the granulocyte series contain the enzyme, esterase, in their granules. This enzyme catalyzes the hydrolysis of the amino acid ester to liberate the pyrrole compound, in turn reacting with the diazonium salt, resulting in a purple color. Nitrite reaction occurs when Nirite reacts with p-arsanilic acid yielding a diazonium compound which, in turn, couples with a quinolin reagent to produce a uniform pink color. Pink spots or edges should not be interpreted as positive. The Specific Gravity test reaction is based on apparent pka change of certain pretreated polyelectrolytes in relation to ionic concentration. In the presence of an indicator, color ranges from deep blue-green in urine of low ionic concentration through green and yellow-green in urines by increasing ionic concentration. This reaction employs 2.8% w/w bromthymol blue, 68.8% w/w/ poly (methylvinylethermaleicanhydride); 28.4% w/w/ sodium hydroxide. Blood reaction is based on the peroxidase-like activity of hemoglobin, which catalyzes the reaction of disopropylbenzene dihyolroperoxide Page 2 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE and uses 3, 3’5, 5’ -tetramethylbenzidine. Blood is detected as intact red cells or as free hemoglobin, although the reagent pad is much more sensitive to the latter. The reaction is based on the liberation of oxygen from peroxide by the peroxidase-like activity of heme. In this test a benzidine compound is oxidized to a colored product ranging from orange to green to dark blue. II. PURPOSE: Bayer Multistix 10 SG and other various Bayer reagent test strips are used as a screening procedure to monitor carbohydrate metabolism, kidney and liver function, acid-base balance and urinary tract functions. It is recommended that a confirmation microscopic exam be performed on the original sample if any of the following are positive: Blood, Protein, Nitrite, Leukocyte Further evaluation or tests may be performed on the original sample if the following are positive: Bilirubin, Ketones III. SCOPE OF PRACTICE: Clinitek 50 urinalysis testing may be performed by VUMC RN’s, LPN’s, Patient Care Partners/Technicians, Operating Room Technicians, and Nursing Externs working in areas/units approved by the POCT Steering Committee to perform point of care urinalysis testing. To maintain this privilege, testing must be performed in accordance with VUMC hospital POCT policy. All staff performing testing must attend POCT orientation, annual proficiency testing, and follow established testing protocol. IV. SPECIMEN REQUIREMENT AND COLLECTION: A. Conditions for Patient Preparation Follow Patient Preparation and Collection Procedure for Point of Care Testing policy. B. Specimen Type At least 12 ml of fresh, first morning urine recommended. C. Handling Conditions Use Standard Precautions Collect urine in a clean, dry container Do not centrifuge Page 3 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE V. Use of preservatives is not recommended. Test urine immediately. Refrigerate if testing cannot be performed within one hour of voiding and let it return to room temperature before testing. Urine with prolonged exposure to room temperature and light may yield altered results for bilirubin, urobilinogen, pH, glucose, and nitrite tests. REAGENTS AND SUPPLIES: Bayer Multistix 10 SG reagent test strips (based on dry weight at time of impregnation) Glucose:2.2% w/w glucose oxidase;1.0% w/w peroxidase; 8.1% w/w potassium iodide; 69.8% w/w buffer; 18.9% w/w nonreactive ingredients Bilirubin: 0.4% w/w 2,4-dichloroaniline diaxonium salt; 37.3% w/w buffer; 62.3% w/w nonreactive ingredients Ketone: 7.1% w/w sodium nitro prusside; 92.9 % w/w buffer Specific Gravity: 2.8% w/w bromthymol blue; 68.8% w/w poly (methyl vinyl ether/maleic anhydride); 28.4%w/w sodium hydroxide Blood: 6.8% w/w diisopropybenzene dihydroperoxide; 4.0% w/w 3,3’,5,-tetramethylbenzidine; 48.0% w/w buffer; 41.2% non-ractive ingredients pH: 0.2% w/w methyl red; 2.8% w/w bromthymol blue; 97.0% w/w inactive ingredients protein: 0.3% w/w tetrabromphenol blue; 97.3% w/w/ buffer; 2.4% w/w nonreactive ingredients Urobilinogen: 0.2% w/w p-diethylaminobenzaldehyde; 99.8% w/w noreactive ingredients Nitrite: 1.4% w/w/ p-arsanilic acid; 1.3% w/w 1,2,3,4tetrahydrobenzo(h)-quinolin-3-ol; 10.8% w/w buffer; 86.5% w/w nonreactive ingredients. Leukocytes: 0.4% w/w derivatized pyrrole amino acid ester; 0.2% w/w diazonium salt; 40.9% w/w buffer; 58.8% w/w nonreactive ingredients BIO-RAD Control System - Urine control Level I and Level II : Human erythrocytes, constituents of animal origin, chemicals, and preservatives. Clinitek 50 Urine Chemistry Analyzer Page 4 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE VI. STORAGE AND STABILITY: VII. Reagent strips must be stored in the original capped vial at room temperature (below 85 degrees F/ 30 degrees C). Replace cap immediately after removing test strip. Opened reagent strips may be used until the printed expiration date on the vial if quality control testing is acceptable. BIO-RAD qUAntify control Levels 1 and 2 will be stable until the manufacturer’s expiration date when unopened controls are stored at 2-8°C (36-46°F). Vials of BIO-RAD qUAntify control Levels 1 and 2 once opened will be stable for 31 days when stored tightly capped at 2-25°C (refrigerated or room temperature).. All reagent strips and BIO-RAD qUAntify control Levels 1 and 2 must be labeled with staff initials and valid expiration date. Do not freeze control solution. Do not store reagent strips in direct sunlight. CALIBRATION: The operator performs no calibration procedure. The Clinitek 50 automatically performs a series of internal checks before and after testing. VIII. QUALITY CONTROL (QC): A “Control” product is used to check the accuracy of the dipsticks, and the technique of the staff member performing the test. After removing control solution from refrigerator allow 15 minutes to reach room temperature before testing. BIO-RAD qUAntify control Levels 1 and 2 must be used to perform a QC test every 24 hours for each vial of test strips in use. BIO-RAD qUAntify control Levels 1 and 2 and Multistix 10 SG test strips may be obtained from Center Supply (materials distribution) A Control Package Insert is included in each box of Control and must be posted in the area where dipstick Control testing is performed. The Expected Ranges using the Multistix 10 SG dipsticks and Level I and Level II Urine Control on the Clinitek 50 must be recorded on the QC log. Control Package Insert Always replace the insert when a new bottle of control is received Lot # on insert must match Lot # on Control Solution bottles Page 5 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE **** CONTROLS, MULTISTIX 10 SG TEST STRIPs , AND THERMAL PAPER MAY BE OBTAINED FROM YOUR SERVICE CENTER OR CENTRAL SUPPLY Urine Control Solution: BIO-RAD Control System Urine Control Level I (NOVA # 36559) and Level II (NOVA # 18143) Printer Paper: CLINITEK 50 THERMAL PAPER (NOVA #30560) (5 ROLLS/BOX) A. Performing Quality Control Test Observe Standard Precautions! 1. Check the expiration date on the control bottle (—expires 31 days after opening) and verify the lot # of the control chart matches the lot # of the control solution. 2. Allow control solution to reach room temperature and Verify the reagent strips are in-date for use. 3. Remove reagent strip and immediately recap the vial. 4. Shake control bottle gently, to mix and remove dropper cap. 5. Invert control bottle and apply control material directly across each pad by gently squeezing the bottle, making sure not to touch tip of bottle to test strip pads and each pad is wet. 6. Press one of the green start buttons on the Clinitek 50 7. Turn test strip on edge to allow excess to drain on an absorbent tissue, or paper towel. 8. Place the test strip on the Clinitek 50 table in the center slot position WITHIN 10 SECONDS of pressing the green start button. 9. The Clinitek 50 will automatically pull the table inside to analyze. 10. Clean dispenser tips and replace cap. 11. Once the test is complete document results and verify all analytes are within expected ranges – found on Control Package Insert. IX. CORRECTIVE ACTION: **If control test is not within range listed on the Control Package Insert: a. Check expiration dates on control bottle and urine reagent strips. If supplies are "out of date", obtain new supplies. If supplies are "in date", repeat test. b. If control test is still out of range, replace control and retest. c. If control test is still out of chart range, replace test strips and retest. d. Indicate corrective action taken on QC log and record the specific problem and action taken on POCT Problem log Page 6 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE e. If troubleshooting steps do not correct the unacceptable control test, follow Clinitek Operator’s Manual troubleshooting steps. If problem is not resolved, contact unit/area manager or POCT Support Center 35707 or 3-8416 X. PERFORMING A PATIENT TEST: 1. Open the urine dipsticks vial and remove one test strip. Immediately recap the vial. 2. Mix patient sample by gently swirling. 3. Dip the test strip into the urine specimen making sure to immerse and saturate all of the test pads. Immediately remove test strip from urine. 4. Press the green start button as the test strip is removed from urine. At the same time Drag the test strip along its edge, across the top of container. 5. Blot the test strip by touching the edge of strip to paper towel. (YOU HAVE 10 SECONDS TO BLOT THE STRIP AND PLACE IT ON THE TABLE) 6. Place the test strip on the table of the Clinitek 50 in the center slot position. The Clinitek will automatically pull the strip holder inside. The result printout is available in 60 seconds. 7. Wipe the strip holder with gauze or paper towel, to remove excess urine. 8. Write the name or MR# of patient on printout and document patient results. See XIII, for documentation requirements. XI. REPORTING RESULTS: Test result names may be abbreviated as shown below. You must include the units for each result where applicable, when reporting patient test results in permanent medical record. The use of symbols (+ /-) are unacceptable when reporting patient or QC results. Page 7 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE See the following chart for printed results: TEST ABBREVIATION UNITS Glucose GLU mg/dL Bilirubin BIL Ketone KET Specific Gravity SG pH pH Protein PRO mg/dL Urobilinogen URO E.U./dL Nitrite NIT Occult Blood BLO Leukocytes LEU XII. ABNORMAL Negative 100 250 NEGATIVE SMALL NEGATIVE TRACE 15 < = 1.005 1.010 1.015 5.0 5.5 6.0 NEGATIVE TRACE 30 0.2 1.0 2.0 500 > = 1000 MODERATE LARGE 40 > = 80 6.5 7.0 7.5 1.020 1.025 > = 1.030 8.0 8.5 > = 9.0 100 > = 300 4.0 > = 8.0 NEGATIVE POSITIVE NEGATIVE TRACE-LYSED TRACE-INTACT NEGATIVE TRACE SMALL SMALL MODERATE LARGE MODERATE LARGE INITIAL CLINITEK 50 SETUP: XIII. mg/dL NORMAL SYSTEM See Clinitek 50 Operator’s Manual (User’s Guide) Changing printer paper, clearing memory etc: Refer to Clinitek 50 setup for instructions. DOCUMENTATION: A. The following quality control information must be recorded on the provided log sheets: Date and time of control testing Testing staff initials Reagent/supplies Lot #, altered Exp. Date, and acceptable QC ranges obtained from the urine control package insert Clinitek 50 serial number Clinitek 50 Control results Daily cleaning of Clinitek 50 table Any Corrective Action Page 8 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE The following information must be recorded in the patient’s chart: B. XIV. If QC is not performed because no patient testing was performed, you must write NPT or no patient testing on QC log for said dates. Patient test results (including units of measurement where applicable) Multistix lot # and expiration date Date and time of patient test Testing staff ID Clinitek 50 serial number All Clinitek Instrument problems and resolutions must be recorded on the provided problem log Storage refrigeration temperatures must be verified daily and recorded on provided temp logs. QUALITY ASSESSMENT The manager or designee has oversight of the following requirements: All POCT performed in patient care areas has been approved by the Diagnostic Laboratories and the State of Tennessee. Testing is performed only by staff approved by the Diagnostic Laboratories and the State of Tennessee. All staff are trained prior to testing, and assessed for competency annually. Individuals responsible for staff training and /or compliance review, (quality control and patient documentation); have attended appropriate training sessions provided by the POCT Support Center. Initial training records, current annual competency records and “designated” trainer records must be accessible. Notification to the POCT Support Center of any changes or transfer of testing personnel. Current testing procedures and accessible to all testing staff. Quality control and maintenance procedures are performed, documented and reviewed for compliance by the manager or designee at least weekly, and are accessible for two years following review. Any corrective action is noted, and filed with records. Routine review of random patient medical records by manager or designee to verify compliance with patient test result documentation as described in testing procedures. Examples must be available for Page 9 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE inspections. All testing supplies are stored and labeled according to instructions found in individual testing procedures. CAP proficiency testing, as required, is performed according to accompanying instructions and results and requested information returned to the POCT Support Center within specified time frame. Diagnostic Laboratories POCT Support Center Provides: Technical support Training Content of training materials and testing procedures Validation and implementation of testing methodologies Assistance with request for testing privileges The POCT Steering Committee is chartered by the VUMC Medical Board and has the authority to revoke testing privileges. XV. INSTRUMENT MAINTENANCE: Observe standard precautions. A. Daily Cleaning The Clinitek 50 test strip table must be kept clean if the instrument is to provide accurate test results and operate correctly. Wipe table with gauze or paper towel after every dipstick test, to remove excess urine. 1. Remove table by pulling it straight out of the instrument. 2. Wet a cotton-tipped stick with distilled water and thoroughly scrub the trough and surrounding areas. Then rinse area well. 3. Dry table well with soft cloth or lint free tissue. 4. Check the white bar for dust, marks, or scratches; if it is dirty wet a cotton-tipped stick and gently wipe the bar. Allow bar to air dry and check it again. If it is scratched or scuffed, or can’t be cleaned contact the POCT Support Center at 3-5707 or 3-8416. **DO NOT USE ANYTHING THAT WILL SCATCH THE WHITE BAR **DO NOT USE SOLVENTS OF ANY KIND TO CLEAN THE BAR Page 10 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE B. Daily Disinfecting the Table 1. Remove, clean, and dry table. 2. Fill a tall, narrow container with about 4 inches of Isopropyl Alcohol or Household Bleach. 3. Put table into solution, making sure the white bar is above the liquid level. 4. Soak the table for no longer than 10 minutes, and then rinse well with water. 5. Dry table with a soft cloth or tissue and insert it back into the instrument. **DO NOT ALLOW SOLUTION TO COME INTO CONTACT WITH THE WHITE BAR** XVI. LIMITATIONS: 1. Drugs that contain azo dyes (e.g. Pyridiium) nitrofurantoin (e. g. Macrodantin), and riboflavin, cause abnormal urine color and may affect the color development on the reagent pad resulting in both false positve and negative results. 2. The pH of the urine can become alkaline on standing due to release of C02 or production of ammonia by urea splitting bacteria. The pH of highly alkaline urine can appear lower due to a “run-over” effect where acid buffer from the protein reagent pad runs over the adjacent pH pad. Highly pigmented urines may mask the color reaction. 3. False negatives at the trace glucose level can be seen when elevated levels of ketone or ascorbic acid are present. 4. False positive proteins may be seen with highly buffered, alkaline urines, exposure to quaternary ammonium compounds, or exposure to skin cleansers containing chlorhexidine. 5. False positive ketone results can be seen in highly pigmented urine specimens and in the presence of high levels of levodopa. 6. False negative bilirubin results may occur with old specimens due to oxidation of bilirubin, and ascorbic acid levels > 25 mg/dL. Indican and other pigmented drugs may interfere with test interpretation. 7. False positive urobilinogen results may occur with para-aminosalicylic acid (a bacterial drug) and sulfonamides. False negatives results may occur when formaldehyde is present. Highly pigmented urines may mask the color reaction. 8. Strong oxidizing agents can cause false positive leukocyte results. Falsely lowered reading may be caused by elevated: 1) glucose ( 3000 mg/dL); 2) specific gravity; 3) protein; 4) various drugs including, Page 11 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE tetracycline, cephalothin, cephalexin and ascorbic acid. Highly pigmented urines, especially those containing azo dyes, may mask this reaction. 9. False negative nitrites may occur with: 1) ascorbic acid levels > 25 mg/dL at low nitrite levels; 2) improper bladder incubation time (< 4 hours); 3) lack of dietary nitrate; 4) high urine specific gravity. 10. Highly buffered alkaline urines may cause low specific gravity readings relative to other methods. Elevated specific gravity readings may be obtained in the presence of moderate quantities (100 - 750 mg/dL) of protein. 11. False positive blood may occur with certain oxidizing contaminants such as hypochlorite and microbial peroxidase. Elevated specific gravity may reduce the reactivity of the test. XVII. LINEARITY: When the Clinitek 50 exceeds the dynamic readout range the following results will be seen: Glucose Bilirubin Ketone Specific Gravity pH Protein Urobilinogen Nitrite Blood Leukocyte SENSITIVITY Glucose Bilirubin Ketone Blood Protein Nitrite Leukocytes 1000 Large 80 1.030 9.0 300 8.0 Positive Large Large 75 – 125 mg/dL 0.4 – 0.8 mg/dL 5 – 10 mg/dL 0.015 – 0.062 mg/dL 15 – 30 md/dL 0.06 – 0.1 mg/dL 5 – 15 cells/hpf in clinical urine Page 12 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE XVIII. PROCEDURAL NOTES: 1. Urine must be collected in a clean dry container free from any detergents, or disinfectant residue since the oxidants contain in such cleaning agents may cause the test areas on the urine test strips for blood and glucose to indicate false positive results. 2. Do not touch the testing area with anything except urine or control solution. 3. Do not use test strip if the testing areas are discolored 4. Do not cut or manipulate test strips. 5. If urine test strip screening is unavailable due to QC, supply or staffing issues, perform visual urinalysis test method or send patient specimens to the clinical laboratory for testing. Differences Between the Human Eye and Instrumental Optics There are inherent differences between the colors that are perceived by the human eye and that are detected by any instrumental optical system. The human eye is capable of detecting minute differences in shade and very small areas of color; artificial optical systems are less sensitive to such small changes. Conversely, instrumental optics is capable of detecting certain colors that are masked by or blended with other colors to the human eye. For this reason, exact agreement between visual results and instrumental results might not be found. However, agreement is generally within one visual color block or reported level and is equal to or better that the agreement between two visual readers. Agreement of urine color is generally within one step along the chromatic scale. XIX. SAFETY: 1. The Clinitek 50 operator should use personal protective equipment (PPE) when performing urine dipstick testing. 2. OSHA and institution protocols for handling bio-hazardous materials should be observed at all times. Page 13 of 14 D:\533582926.doc VANDERBILT UNIVERSTIY MEDICAL CENTER POINT OF CARE TESTING CLINITEK 50 URINALYSIS PROCEDURE XX. REFERENCES: 1. Bayer Corporation Diagnostic Division, Elkhart, Indiana, Clinitek 50 User’s Guide, July 1996. 2. Siemens: MULTISTIX10SG Product Insert # AN30516F, Siemens Healthcare Diagnostics Inc., Tarrytown, NY 10591-5097 USA. Rev. 08/08. 3. BIO-RAD: Control System product insert #02A-975. BIO-RAD Laboratories, Clinical Diagnostics Group, 9500 Jeronimo Road, Irvine, California 92618. Rev 8-2008. 4. VUMC Hematology- Urine Reagent strip Testing Procedure, 2002 5. VUMC POCT Support Center Page 14 of 14 D:\533582926.doc