HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

APPROVED BY:

.. ASSOC PROF DR ROSLINE HASSAN HEAD OF HAEMATOLOGY DEPARTMENT

CONTROLLED COPY NO : 3 REGISTERED HOLDER HAEMATOLOGY LABORATORY

RECORD OF REVIEW/AMMENDMENT DATE 1.8.2011 VERSION NO. DETAIL OF AMMENDMENT Add test procedure for a new machine STA-R Evolution (5.1b) BY Wan Soriany bt Wan Md Zain

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

PREPARED BY DESIGNATION CHECKED BY DESIGNATION AUTHORISED BY DESIGNATION 1. OBJECTIVE Quantitative Determination of D-Dimer in human citrate plasma 2. METHOD The detection of latex immunoassays is based on the absorbance (optical density: O.D.) of monochromatic light at 540nm. A antigen-antibody reaction takes place respectively when monochromatic light passing through the cuvette. 3. PRINCIPLE Quantitative determination of D-Dimer by the immuno-turbidimetric method. This assay is based on the change in turbidity of a microparticles suspension that is measured by photometry at 540nm. A suspension of latex microparticles, coated by covalent bonding with monoclonal antibodies specific for D-Dimer, is mixed with the test plasma whose DDimer level is to be assayed. An antigen-antibody reaction takes place, leading to an agglutination of the latex microparticles that induces an increase in turbidity of the reaction medium. This increase in turbidity is reflected by an increase in absorbance. The increase in absorbance is a function of the D-Dimer level present in the sample. 4. REQUIREMENTS 4.1 EQUIPMENT 4.1.1 4.1.2 4.1.3 4.1.4 STA Compact or STA-R Evolution Centrifuge Micropipettes (500 l, 1000 l) Pipette tips

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

: MASETA ISMAIL : TECHNICAL OFFICER / MED LABORATORY TECHNOLOGIST U32 : ASSOC PROF DR WAN ZAIDAH WAN ABDULLAH : DEPUTY QUALITY MANAGER / HAEMATOLOGIST : ASSOC PROF DR ROSLINE HASSAN : HAEMATOLOGIST / LAB DIRECTOR

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

4.2 REAGENT 4.2.1 STALiatest D-Dimer REF 00515 Reagent 1: Tris Buffer; Reagent 2: Suspension of microlatex particles coated with two different mouse monoclonal anti-human D-dimer antibodies. 1. Allow the Reagent 1 and reagent 2 to stand at room temperatures (18 25 C) for 15 minutes before use. . 2. Mix the reagents by gentle swirling of the vials without creating any bubbles. 3. On STA Compact stable for 15 days. 4.2.2 STAOwren Koller (REF 00360) 1. If the buffer is refrigerated allow it to stand at room temperature (18 25 C) for 30 minutes before use. 2. On STA Compact stable for 3 days.

4.3 CONTROL STA-Liatest Control N + P (REF 00526) 1. Reconstitute each vial with 1 ml of distilled water. 2. Allow the reconstituted material to stand at room temperatures (18 25 C) for 30 minutes. 3. Then swirl the vial gently before use. 4. On STA Compact stable for 8 hours.

4.4

SPECIMEN COLLECTION AND TREATMENT Blood (9 volume) is collected in 0.109M (i.e. 3.2%) trisodium citrate anticoagulant (1 volume) Centrifugation: 15 minutes at 2000-2500g Plasma stability: 8 hours at 20 + 5oC 1 month at -20 o C Frozen plasma must be thawed directly at 37 o C for 15 min before testing.

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

5. 1a NO 5.1

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

PROCEDURE BY STA COMPACT ACTIVITY RESPONSIBILITY MLT/SO Enter

Run calibration when reagent lot number changed @ QC result not good Esc Main Menu Calib/ Control Enter Calibration

5.2

Notes : Review calibration. Shall perform and review a calibration* whenever reagent lot number changed or QC result not satisfactory. * If calibration invalid r 0.985, rerun the calibration. Perform Quality Control Esc Main Menu Calib/ Control Enter Quality Control Enter

MLT/SO

5.3

Notes: Review quality control data. If QC out of range, rerun the QC. (Refer to Westgard et al for identification and resolution of out of-control situations). If QC out of range, rerun the QC Loading Reagent@Control In Product Drawer 5.3.1 5.3.2 5.3.3 5.3.4 Open product drawer ( Press F2) Scan barcode label of reagent bottle Press Enter (the cursor will move to Pos) Put the bottle in the drawer ( ensure a beep sound is heard) If the lot number of reagent change is detected, the following window is displayed:
LOT NUMBER CHANGED All concerned calibrations will be definitely invalid Accept (YES/NO)

i. ii. iii. 5.3.5 5.3.6 5.3.7 5.3.8 5.3.9 5.3.10 5.3.11 5.3.12 5.3.13 5.3.14 5.3.15

Type YES and press ENTER Scan the barcode sheet Press ESC when end of data processing

Continue loading STA-Liatest Control N+P Close the Product Drawer when end of loading Open Sample drawer (press F1) Scan barcode label of STA-Owren Koller bottle Choose F1 for Diluent Press ENTER (the cursor will move to Pos) Put the bottle in position 8 (ensure the beep sound is heard) Close the Sample Drawer when end of loading If QC results OK, proceed with patient testing Open Sample Drawer #Press F12 if the patient sample is STAT sample
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

5.3.16 5.3.17 5.3.18 5.3.19 5.3.20 5.4

#Press F8 to change Normal Tube to Micro Tube if the paediatric sample tube has been transferred to adaptor Key in Patient ID @ Scan barcode label Press ENTER (the cursor will move to Pos) Put the sample tube in the drawer (#ensure a beep sound is heard) Select the test (Press 1,2, @3 according to Test Profile) Press F10 to validate MLT/SO

Loading sample In sample Drawer 5.4.1 Open Sample Drawer 5.4.2 #Press F12 if the patient sample is STAT sample #Press F8 to change Normal Tube to Micro Tube if the pediatric sample tube has been transferred to adaptor 5.4.3 Enter Patient ID or Scan barcode label on tube. 5.4.4 Put the sample tube in the drawer (#ensure a beep sound is heard) 5.4.5 Select the test (D-Dimer) 5.4.6 Press F10 to validate 5.4.7 Continue loading other patient samples. If not, close the Sample Drawer 5.4.8 Press ESC 2 times, test panel screen will appear. 5.4.9 Select barcode number, press ENTER, Test Panel - File Processing Screen appear move cursor to green box. 5.4.10 Key in Patients name, RN and MLT. Move cursor to test selected, press F10 to save.

5.5 5.5

Result Entry 1. The result is entered on the appropriate Request Form 2. Record the result in Laboratory Information System (LIS) Result Print-Out 1. Press Esc Main Menu Files Enter Confirmation/printout Enter

MLT/SO MLT/SO

2. CONFIRMATION/PRINTOUT screen displayed, Press the ESC key , the window displayed: Change File Selection Transmit Print Quit With cursor positioned on Quit 3. Move the cursor to Print by pressing the key, then confirm with the The PRINTOUT window is displayed , see description below key.

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

PRINTOUT Unprinted Files All Files Printer Setup Quit Depending on the required patient files: Quit Move the cursor to all files or Unprinted Files, then confirm with the key. The file printout starts.

5. 1b NO 5.1 5.1 5.1.1

PROCEDURE BY STA-R EVOLUTION ACTIVITY Loading Reagent @ Control in Product Drawer To Open Product Drawer icon RESPONSIBILITY

1. From Functional Panel menu, click on 2. Click on OPEN tab

5.1.2 Loading a reagent or control with bar code identification For STA bar coded reagent only 1. Open Product Drawer 2. Scan bar code label of reagent or control vial through build-in barcode reader 3. Check the box or press F8 key for microvolume sample ** Reagent or control must transfer into microcups ** Change the reagent or control volume if necessary 4. Check reagent appropriate position availability by clicking the down arrow ** R0 only reserved for calibrator, control, diluent and Desorb U ** R1 only reserved for intermediate reagent and Desorb U ** R2 only reserved for Start reagent and Desorb U 5. Load reagent or control into appropriate position 6. If different lot number of reagent/control/calibrator is detected, following Question displayed Do you wish to read the calibration parameters using
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

the barcodes? Click on OK tab to read the reagent or control bar coded sheet immediately. Please proceed to step 10 to step 14. OR Click on Abort tab to read the reagent or control bar coded sheet after close the Product drawer. Please proceed to step 7 to step 12. 7. Continue loading of next reagent or control 8. Click on Close Drawer tab to close product drawer. 9. In the Product Screen menu, click on ** 583 represented last 3 digits of new reagent lot. 10. Scan reagent or control bar coded sheet 11. Click on Confirm tab 12. Click on Close and Calibrate to start the calibration 13. Continue loading of next reagent or control 14. Click on Close Drawer tab to close product drawer. 5.1.3 Loading a reagent or control using manual identification For non STA bar coded reagent 1. Open Product Drawer 2. Type the product identification as defined in the Test setup menu, then click on to confirm Example: ID for STA PTT Automate 5 is 12203 3. Check the box or press F8 key for microvolume sample in Lot column

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

** Reagent or control must transfer into microcups ** Change the reagent or control volume if necessary 4. Type the reagent or control lot number, then click on to confirm

5. Check reagent appropriate position availability by clicking the down arrow ** R0 only reserved for calibrator, control, diluent and Desorb U ** R1 only reserved for intermediate reagent and Desorb U ** R2 only reserved for Start reagent and Desorb U 6. Load reagent or control into appropriate position
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

7. If different lot number of reagent/control/calibrator is detected, following Question displayed Do you wish to read the calibration parameters using the barcodes? Click on OK tab to read the reagent or control bar coded sheet immediately. Please proceed to step 10 to step 14. OR Click on Abort tab to read the reagent or control bar coded sheet after close the Product drawer. Please proceed to step 7 to step 12. 7. Continue loading of next reagent or control 8. Click on Close Drawer tab to close product drawer. 9. In the Product Screen menu, click on ** 583 represented last 3 digits of new reagent lot. 10. Scan reagent or control bar coded sheet 11. Click on Validate tab 12. Click on Close and Calibrate to start the calibration 13. Continue loading of next reagent or control 14. Click on Close Drawer tab to close product drawer. CALIBRATION Calibration should be performed when reagent lot change. Pre-calibrated test Only 2 active calibrations per test (2 different lots) allowed. The oldest calibration curve will automatically delete when third reagent lot introduced to system 1. From the message box Do you wish to read the calibration parameters using the barcodes? 2. 3. Click on OK tab Scan reagent or Control bar code sheet MLT/SO in Lot column

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

4. 5. 6. 7.

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

Click on Confirm tab Click on Close and Calibrate tab Click on Validate tab Enter Password and User Identification

Notes : Review calibration. Shall perform and review a calibration* whenever reagent lot number changed or QC result not satisfactory. * If calibration invalid r 0.985, 5.2 Perform Quality Control 5.2.1 Multiple Test Controls Request 1. From Function Panel menu, click on 2. Highlight the test, e.g. PT. 3. Continues selection of other tests, e.g. APTT, FIB and D-Dimer by using Shift key or Ctrl key for random selection 4. Click on icon icon rerun the calibration MLT/SO

5. Enter Password and User Identification 6. Click on OK tab 5.2.2 Single Level Control Test Request 1. From Function Panel menu, click on 2. Highlight the test, e.g. PT. 3. Click Control level 1 or Control level 2 tabs 4. Click on icon icon

5. Enter Password and User Identification 6. The following message displayed: PT Level: 1. Run the control? 7. Click on OK tab

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

5.2.3

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

To View Quality Control Result

To View Daily Quality Control Result in Tabular Mode 1. From Function Panel menu, Click on 2. Click on icon icon

3. Daily Control Screen open and daily quality control results displayed. 4. Re-sort the daily control displayed sequence by test, control level, result, etc by clicking on the heading of the column you would like to use. 5. If control is out or to be validate, double click or right click on the control in the Status column. Click on Validate, Delete or Rerun to validate, delete or rerun the control. 5.2.4 To view Daily Quality Control in Graphic Mode icon

1. From Function Panel menu, Click on 2. Highlight the test, e.g. PT.

3. Click Control level 1 or Control level 2 tabs to view QC graph Notes: Review quality control data.If QC out of range, rerun the QC. (Refer to Westgard et al for identification and resolution of out of-control situations). 5.3.1 Loading Sample Tubes 1. From Function Panel menu, click on 2. Click on Test Panel tab 3. Make sure either screen. 4. Load sample tubes into the sample rack 5. Load sample racks onto sample rack tray. 6. Load sample rack tray onto the Rack Tray Loading Zone (left side of STA-R) 7. The sample tubes automatically loaded and sample tube bar code label automatically scan and identify. 8. If the sample tube bar code label unable to read, the sample rack will unload and following message appear:
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MLT/SO icon

icon or

icon are presented in the Test Panel

HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

There are unread IDs in the rack. Please re-enter the IDs 9. Click on OK tab 10. Manual Input of Patient IDs screen appear for sample tube re-identification

1. Check the unread sample tube position. Click on unread sample tube to be enter by clicking the number of tube position on 2. Enter the sample tube bar code manually on STAT for urgent sample on Pediatric for microvolume sample.

3. Click on 5.3.2 Request Test

to confirm.

5.3.2a: Request Test by Profile 1. From Function Panel menu, click on 2. Click on Test Panel tab 3. Tag sample to be add test or Click on T column heading to Tag All sample 4. Click on Apply Profile to Tagged Identities 5. Choose and click on the profile from the list 5.3.2b: Request Single Test 1. From Function Panel menu, click on 2. Click on Test Panel tab 3. Tag sample to be add test or Click on T column to Tag All sample 4. Click on Test Abbreviation header, e.g. PT, Click on Add tab 5. Add Test for all Tagged Identities? Test: PT? message appear 6. Click on OK tab
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icon

icon

HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

5.3.2c: Sample with Automatic Profile 1. From Function Panel menu, click on 2. Click on (Un) Loading tab 3. on Add Automatic Profile 4. Loading sample rack 5. The tests will automatically add and perform 5.3.2d Loading STAT Sample 1. From Function Panel, click on 2. Please make sure there is no rack in loading position before ending of Initialization message appear 3. Click on OK tab 4. Initialization of rack loading system. Wait until The message Load a new rack inside the instrument is displayed. 5. Push rack into the STA-R Evolution analyzer with rack barcode face inside 6. Rack load into sample area 7. Tag STAT sample 8. Click on Test Abbreviation header, e.g. PT, Click on Add tab to add single Test OR Click on T column heading, Click on Apply Profile to Tagged Identities and Choose and click on the profile from the list 5.4 View Patient Result a.View Non-Archived Patient Result ** Sample test request completely processed and tube remain in the analyzer. 1. From Function Panel menu, click on icon. MLT/SO icon

2. Click on Test Panel tab, ten patient files are displayed. 3. Use the horizontal scroll bar to view other patient results. 4. Click on icon, select Search By tab and choosing the search method by

Clicking on Identity, Name or First Name tab 5. Click on Search tab. 6. Click or enter key to confirm.
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4 VERSION NO: VERSION DATE: 1 01.11.2009

b. View Archived Result ** Sample test request completely processed and tube removed from the analyzer. 1. From Function Panel menu, click on icon.

2.

Click on

icon.

3. 4.

Click on

icon.

Select Search By tab and choosing the search method by clicking on Identity or Date tab.

5. 6.

Click on Identity tab, click on Search tab for the entire day result. OR Click on Date tab, change the date for the period of the result to be view. Click on Search tab.

5.5

Printing Patient Result a. Printing Non-Archived Patient Result

MLT/SO

1. 2. 3. 4. 5.

From Function Panel menu, click on Tag the patient file to be printed. Click on icon

icon.

The following message displayed: Print All Tagged Files? Click to confirm.

b. Printing Archived Patient Result

1.

From Function Panel menu, click on

icon.

1.

Click on

icon.

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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

2. 3. 4. 5. Click

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

Tag the patient file to be printed. Click on icon or

Click the T column heading, select Printed All Tagged Files The following message displayed: Print All Tagged Files? to confirm.

6.

LIMITATION 6.1 6.2 6.3 6.4 6.5 Cloudy plasmas may lead to an under-estimation of the D-Dimer level. Ensure that the absorbance value at 540nm of the plasma diluted 1:6 with STA-Owren Koller is <0.35 Concentrations of fibrinogen degradation product greater than 15 g/ml may lead to an over-estimation of the D-Dimer level. The presence of rheumatoid factor at a level greater than 50 IU/ml may lead to an over-estimation of the D-Dimer level. The STA Liatest D-Di is insensitive to the following substances: hemoglobin (up to 5 g/l), bilirubin (up to 200 mg/l), unfractionated heparin (up to 2IU/ml), and low molecular weight heparin (up to 2 anti Xa IU/ml). The presence of anti-bovine albumin and/or anti-mouse antibodies in certain subjects may lead to an over-estimation of D-Dimer level.

7.

REFERENCE RANGE The normal level of D-Dimer in adult population is generally < 0.45 g/ml (expressed FEU).

8.

RESULT The D-Dimer level is expressed as g/ml (FEU). FEU is fibrinogen equivalent units in which one FEU is the quantity of fibrinogen initially present that leads to the observed level of D-Dimer. The actual quantity of D-Dimer is approximately half of an FEU. The cut-off value of 0.50g/ml is approximately 0.25g/ml actual D-Dimer.

9.

REFERENCES 1. 2. 3. 4. 5. Diagnostica Stago Haemostasis brochure MHT/DL/102 STA Compact Protocols- Tests Settings MHT/DL/106 STA Compact User Operation & Training Manual MHT/DL/109 STA Compact Operators Manual MHT/DL/110 Insert package STA- Liatest D-DI November 2005
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO.

6. 7.

DETERMINATION OF D-DIMER (AUTOMATED) HUSM/HEMA-UPT/STM-C4

VERSION NO: VERSION DATE:

1 01.11.2009

National Committee for Clinical Laboratory Standards. Preparation and Testing of Reagent Water in the Clinical Laboratory, Third Edition, NCCLS Document C3-A3: Vol. 17 No.18 Guideline: Quality Assurance Of Coagulation Laboratory

END OF DOCUMENT

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