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DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
APPROVED BY:
RECORD OF REVIEW/AMMENDMENT DATE . 1.8.2011 VERSION NO. . DETAIL OF AMMENDMENT Sections on Limitations have been change to Sources of errors (section 6.0). Add test procedure for a new machine STA-R Evolution (5.1b) . Wan Soriany bt Wan Md Zain BY
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DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1
1 01.11.2009
: WAN SORIANY WAN MD ZAIN : QUALITY OFFICER / SCIENTIFIC OFFICER : ASSOC PROF DR WAN ZAIDAH WAN ABDULLAH : DEPUTY QUALITY MANAGER / HAEMATOLOGIST : ASSOC PROF DR ROSLINE HASSAN : HAEMATOLOGIST / LAB DIRECTOR
REAGENT 4.2.1 PT Reagent Reagent 1STA Neoplastine CI Plus (REF 00667 or 00606)
STANDARD TECHNICAL MANUAL
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
Reagent 2 (Solvent) 1. Transfer the entire contents of one vial of Reagent 2 (R2) into one vial of Reagent 1 (R1) of the same kit. 2. Allow the reconstituted reagent to stand at room temperature (18 -25 C) for 30 minutes. 3. Swirl the Reagent 1 vial gently to obtain a homogenous suspension. 4. Once reconstituted the Reagent 1, with stirring-bar, remains stable for 48 hours on STA analyzer (15 - 20 C) 4.2.2 APTT Reagent STA PTT A (REF 00595) 1. Reconstitute each vial with 5ml of distilled water. 2. Allow the reconstituted material to stand at room temperature for 30 minutes (18 -25 C). 3. Swirl the reagent vial gently to obtain a homogenous suspension. 4. Once reconstituted the reagent remains stable for 24 hours on STA Compact STA CACl2 0.025 M (REF 00367) 1. If the reagent is refrigerated, allow it to stand at room temperature (18 -25 C) for 30 minutes, before use 2. After opening, remains stable for: 24 hours at 37 C 3 days on STA Compact 4.3 CONTROL STA- Coag Control N + P (REF 00679) Reagent 1 (STA Coag Control N) Reagent 2 (STA Coag Control P) 1. Reconstitute each vial of Reagent 1 and Reagent 2 with 1 ml distilled water. 2. Allow the reconstituted material to stand at room temperature (18 -25 C) for 30 minutes. 3. Swirl the vial gently before use. 4. Once reconstituted, Reagent 1 and 2 remain stable for 8 hours on STA analyzer. 4.4 SPECIMEN COLLECTION AND TREATMENT i) Blood (9 volume) is collected in 0.109 M (i.e 3.2%) trisodium citrate anticoagulant (1 volume) Centrifugation: 15 min at 2.500 g PT: Plasma stability: 8 hours at 20 5 C Do not store at 2-8C (cold activation of factor Vll)
APTT: Plasma stability 4 hours at 20 5C 2 hours if on heparin therapy 4 hours if in CTAD tubes to prevent heparin inactivation
Page 3 of 17 STANDARD TECHNICAL MANUAL
HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
ii)
Monitoring heparin
When monitoring heparin therapy, any release of platelet factor 4 which is a potent inhibitor of heparin, represents a major source of error. Perform centrifugation within 1 hour if the blood was collected in conventional citrate anticoagulant and within 4 hours if the blood was collected with CTAD tubes. iii) High PCV (>55 %)
The amount of citrate must be adjusted (decreased) in patients who have haematocrit values above 55%. The formula below is used to calculate the amount of anticoagulant required:X = [ (100 PCV) / (595 PCV)] Vol Example: If a patient has a hematocrit of 60%, to determine the volume required for 5 mL anticoagulated blood, X = [ (100 60) / (595 60)] 5 = 0.37mL . 5. 1a NO 5.1 PROCEDURE (PT & APTT) by STA-Compact ACTIVITY Run calibration when reagent lot number changed @ QC result not good. Press Esc Main Menu Calib/ Control Enter Calibration Enter RESPONSIBILITY MLT/SO
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 5.2 Perform Quality Control. Press Esc Main Menu Calib/ Control Enter Quality Control Enter MLT/SO
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 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 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
Page 4 of 17 STANDARD TECHNICAL MANUAL
MLT/SO
DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1
1 01.11.2009
Type YES and press ENTER Scan the barcode sheet Press ESC when end of data processing
Continue loading other reagents @ control If not ,close the Product Drawer If QC results OK, proceed with patient testing
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 (PT, aPTT) 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
Result Entry 1. 2. The result is entered on the appropriate Request Form Record the result in Laboratory Information System (LIS)
MLT/SO
Note: Result with panic value (informed immediately) PT: > 40 sec INR: > 5 (on warfarin) APTT: > 100 secs 5.6 Result Print-Out 1. Press Esc Main Menu Files Enter Confirmation/printout Enter MLT/SO
HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
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
PRINTOUT Unprinted Files All Files Printer Setup Quit
Quit
key.
Depending on the required patient files: Move the cursor to all files or Unprinted Files, then confirm with the key. The file printout starts.
Quit
NO 5.2.1
5.2. PROCEDURE FOR PT OR APTT MIX ACTIVITY Immediate mixing for PT and aPTT Prepare 1ml normal pool plasma in an eppendorf tube with adaptor. 1. Open sample Drawer. 2. Type manually the below information: ID: POOL F1 for diluents Name: NORMAL Vial: 1 ml Stability: 4 h 3. Press Enter 4. Put pool plasma tube in any position. 5. Patient ID and put patients plasma in any position 6. Choose PT Mix or APTT Mix 7. Press F10
RESPONSIBILITY MLT/SO
5.2.2
Incubation mixing test for PT and aPTT (2 hours incubation at 37oC) 1. Prepare 1ml normal pool plasma, 1ml patient plasma and 1ml of mixed 1:1 of normal pool plasma and patient plasma in an eppendorf tube and incubate for 2 hours at 37oC. 2. *Prepare (500ul of incubated normal pool plasma and 500ul of incubated
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MLT/SO
HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
patient plasma) in an eppendorf tube. * this tube is refer to then mix in the mixing test worksheet 3. Run the sample as aPTT test or PT test.
PROCEDURE (PT & APTT) by STA-R Evolution ACTIVITY Loading Reagent @ Control in Product Drawer To Open Product Drawer icon RESPONSIBILITY MLT/SO
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 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
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
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
** 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 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.
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
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. Only 2 active calibrations per test (2 different lots) allowed. The oldest calibration curve will automatically delete when third reagent lot introduced to system. Reference Pool clotting time / Reference Time is corresponds to the clotting time of pool of normal plasma, which is pool from at least 20 fresh healthy donors. OR else commercial pool normal plasma, Pool Norm (REF 00539) can be used. MLT/SO in Lot column
1.
2. 3.
Select appropriate reagent lot by clicking on Click on Rd for PT and Ra for APTT to check and confirm on reagent lot to calibrate.
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DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1
1 01.11.2009
Double click on PT or APTT Click on Calibrate tab Enter Password ( USM) and User Identification ( e.g. LING) Click on Ra for APTT and Rd for PT, and select the correct reagent lot to be calibrate. Make sure reagent lot writing in green color.
8. 9.
10. Click on Validate tab OR 11. From the message box Do you wish to read the calibration parameters using the barcodes? 12. Click on OK tab 13. Scan reagent or Control bar code sheet 14. Click on Confirm tab 15. Click on Close and Calibrate tab 16. Enter Password and User Identification 17. Enter Reference Time value 18. Click on Validate tab 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 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
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MLT/SO
icon
DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1
1 01.11.2009
4. Click on
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 5.2.3 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 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
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
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
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: 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 icon or icon are presented in the Test Panel icon
MLT/SO
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
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
3. Click on 5.3.2
to confirm
Request Test
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 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
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
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
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.
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.
DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1
1 01.11.2009
5.5
Click on Date tab, change the date for the period of the result to be view. Click on Search tab. 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.
1.
icon.
2. 3. 4. 5. 6. 7.
Click on
icon.
Click the T column heading, select Printed All Tagged Files The following message displayed: Print All Tagged Files? Click to confirm.
6.
HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
Overfill or underfill of collection tubes Failure to correct the citrate volume for persons with high PCV (>55 %) Incorrect volume, type or concentration of anticoagulant in the collection tube. Clotted, haemolysed sample Inadequate or too vigorous mixing of the specimen with the reagents Contamination collection or storage tubes Contamination with heparin(Note: STA-Neoplastine CI Plus is insensitive to unfractionated heparin levels up to 1 IU/ml and to low molecular weight heparin levels up to 1.5 anti-Xa IU/ml. v) Improper or defective specimen collection tubes vi) Do not keep plasmas at 2-8 C because FVII may be activated by the kallikrein system. vii) Thrombin Inhibitors (Note: Presence of thrombin inhibitors (e.g. hirudin, argatroban) may lead to prolonged prothrombin time. 6.2) Reagent related problems i) Defects in the reagent due to mishandling in shipping or storage ii) Use of reagent beyond the stated reconstitution stability date or beyond the expiration date. 6.3) Analytical errors i) Inaccurate or imprecise dispensing of reagents ii) Instrument malfunction, such as defective bulb, incorrect temperature, reagent splash, poor reagent delivery, or electrical interference. 7. REFERENCE RANGE Normal range INR: 0.86 1.14 Normal range APTT: 30.0 45.8 sec 8. RESULT The PT results can be expressed in different units: As a time (secs) As a Ratio As a INR = Patients PT ISI Mean Normal PT The APTT results can be expressed in different units: As a time (secs) As a Ratio =
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HAEMATOLOGY DEPARTMENT, HOSPITAL UNIVERSITI SAINS MALAYSIA TITLE: PROCEDURE NO. DETERMINATION OF PROTHROMBIN TIME (PT)& ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT) HUSM/HEMA-UPT/STM-C1 VERSION NO. VERSION DATE. 1 01.11.2009
Report the results on-line (by LIS), dispatch the original form and inform results bu phone for the panic result or if request by ward. 9. INTERPRETATION 1. The PT is commonly used for monitoring oral anticoagulant (warfarin) therapy because of its sensitivity to variations in the concentration of the vitamin K dependent factors ll, Vll and X. 2. The APTT is commonly used for monitoring heparin therapy. 3. Prolonged APTT clotting times may be observed in the following situations: deficiency for factor XII, XI, X, IX, VIII, V, II, or fibrinogen, liver diseases, Vitamin K deficiency, presence of heparin, lupus anticoagulant or other inhibitor.
10. 1. 2. 3. 4. 5. 6. 7. 8. 9.
REFERENCES 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- Neoplastine CI Plus Revised October 2003 Insert package STA- PTT A Revised September 2004 Insert package STA- Coag Control N + P Revised March 2005 Insert package STA- CaCl2 Control N + P Revised September 2004
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 10. Quality Assurance For Coagulation Lab. HUSM/58/PK/80 11. Collection of Coagulation Specimen. HUSM/58/PK/88 END OF DOCUMENT
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