Policy # MI_SER_GAL Department of Microbiology. Page Quality Manual TABLE OF CONTENTS

Transcription

1 Department of Microbiology Version: 1.0 CURRENT 1 of 23 Prepared by QA Committee Issued by: Laboratory Manager Revision Date: 3/26/2018 Approved by Laboratory Director: Annual Review Date: 9/1/2018 Microbiologist-in-Chief Uncontrolled When Printed TABLE OF CONTENTS Introduction... 2 Specimen Collection and Processing... 3 Materials and Reagent... 2 Procedure... 2 System Reagent Preparation... 4 Specimen Treatment... 4 Testing Procedure... 5 Reporting Trouble shooting Maintenance Procedure Weekly Monthly Record of Edited Revisions... 22

2 Version: 1.0 CURRENT 2 of 23 Introduction The Platelia TM Aspergillus EIA is a qualitative enzyme immunoassay for the detection of Aspergillus galactomannan antigen in serum and Bronchoalveolar Lavage (BAL) samples. Reagent And Material 1. EVOLIS Analyzer 2. BioRad Aspergillus Galactomannan kit Store the kit at 2-8ºC. Bring all reagents to room temperature(18-25 ºC)for at least 30 minutes before use. Return all reagents to 2-8 ºC immediately after use. Return unused strips/plates to pouch and reseal. Galactomannan monoclonal antibodies coated microwell plate Concentrated wash solution (20X) Negative control serum (Human) Cut-off control serum (Human) Positive control serum (Human) Conjugate Anti-galactomannan monoclonal antibody/ peroxidase labeled Sample Treatment Solution (EDTA acid solution) Chromogen TMB solution Stopping solution 1N sulphuric acid 3. Distilled or deionized water, for dilution of Concentrated Washing Solution 4. Disposable gloves 5. Pipettes to measure and dispense 50 µl, 100 µl, 300 µl, and 1000 µl ml polypropylene microcentrifuge tubes with airtight stoppers, able to support heating to 120 ºC (heat block) 7. Heat block 8. Votex agitator Reagent Preparation And Storage 1. Microwell Strip Plate (R1) Every frame containing 12 strips I s packaged in a pouch. Open the pouch and take out the frame. Put the frame containing the unused strips back in the original pouch. Carefully reseal the pouch and store at 2-8ºC. After the vacuum-packed pouch has been opened, the strips stored at 2-8ºC in their original pouch that has been carefully resealed are stable for 8 weeks. Check whether the desiccant is still present.

3 Version: 1.0 CURRENT 3 of Washing Solution (R2) Prepare Working Washing Solution as neede by adding one part Concentrated Washing Solution (R2) to 19 parts deionized or distilled water. The Working Washing Solution can be stored for 14 days at 2-30 ºC. Prepare a sufficient amount of Working Washing Solution to complete the run (80ml for one strip: 4 ml R ml distilled water). 3. Negative Control Serum (R3), Cut-off Control Serum (R4) and Positive Control Serum (R5) The controls must be heat-treated with the Sample Treatment Solution (R7) as patient specimens, in order to also be a monitor of the treatment. At least one set of controls is needed for each batch of heating process. After opening, these reagents stored at 2-8ºC, are stable for 8 weeks, in the absence of contamination. 4. Conjugate (R6), Sample Treatment Solution (R7), Chromogen: TMB solution (R9) These reagents are ready to use. After opening, these reagents stored at 2-8ºC are stable for 8 weeks if they are free of contamination. 5. Stopping reaction (R10) This reagent is ready to use. After opening, this reagent stored at 2-8ºC is stable until the validity date shown on the label if there is no contamination. Specimen Collection and Processing The test is performed on serum or BAL fluid 1. Serum Collect blood samples according to standard laboratory procedures. Serum samples must be uncontaminated with fungal spores and/or bacteria. Transport and store samples in sealed tubes, unexposed to air. Unopened samples can be stored at 2-8ºC for up to 5 days prior to testing. After initial opening, samples may be stored at 2-8ºC for 48 hours prior to testing. For longer storage, store the serum at -70 ºC. Serum samples can be subjected to a maximum 4 freezing/thawing cycles. Previously frozen specimens should be thoroughly mixed after thawing prior to testing. The results are not affected by sample containing 20mg/L of bilirubin, lipemic samples containing the equivalent of 2 g/l of triolein (triglyceride) or hemolyzed samples containing 500 mg/dl of hemoglobin. Do not decomplement sera

4 Version: 1.0 CURRENT 4 of BAL Fluid Collect BAL fluid damples according to standard laboratory procedures. BAL fluid samples must be collected in sterile saline and may be tested on neat samples (as is) or supernatants from centrifuged samples (10,000 rpm for 10 min) before proceeding to treat the sample. BAL fluid samples must be uncontaminated with fungal spores and/or bacteria. Transport and store samples in sealed tubes, unexposed air. After initial opening, samples may be stored at 2-8ºC for up to 24 hours. For longer storage, store the BAL samples frozen (-20 ºC or less) up to 5 months. BAL samples can be subjected to a maximum of 4 freezing/thawing cycles. Previously frozen specimens should be thoroughly mixed after thawing prior to testing. Procedure System Reagent Preparation 1. Before beginning EIA assay, fill up Galactomannan BioRad Wash containers when necessary: *Working Galactomannan Wash buffer: Add 50 ml of 20x BioRad wash solution to 950 ml of distilled water. Fill up Galactomannan wash container with black tubing 2. De-ionized Water: Fill up Evolis blue, yellow and red tubing container with de-ionized water 3. The Aspergillus Galactomannan conjugate, TMB and stopping solution will be pipetted from their original containers and can be loaded onto the analyzer as is. 4. Bring all required microwell strips and the reagents mentioned above to room temperature before use. 5. Empty the waste tank and check the system liquid container (refill if necessary). System Liquid : 10L of de-ionized water only Specimen Treatment Separate BAL and serum and according to worklist Serum and BAL samples need to be heat-treated in the presence of EDTA prior to being tested on the analyzer. This pre-analytical procedure helps dissociate immune complexes and causes serum proteins to precipitate which can possibly interfere with the testing procedure. All control sera: negative (R3), cut-off (R4) and positive (R5) must be processed at the same time as serum / BAL fluid samples: Each batch of heating process needs a set of controls to be processed at the same time.

5 Version: 1.0 CURRENT 5 of Label 2 empty 2ml sample tubes. One tube must labelled with the patient barcode label. For the control samples label 2 empty tubes for each: Neg, Cut-Off & Pos, respectively. *The Cut-Off Control will be sampled twice by analyzer but only one sample tube needs to be prepared. 2. Pipette 390ul of each test serum/bal or control into one of the 2 ml sample tubes. 3. Add 130ul of serum treatment solution into the sample tube. 4. Tightly close the caps of the tubes to prevent opening during heating. 5. Mix tubes thoroughly by vortexing each sample. 6. Heat the sample tubes by placing them in a heating block at 120 o C for 6 minutes. Tubes must be placed in the block only when the prescribed temperature is reached. Do not rely on the temperature displayed by the apparatus, please check that the emperature complies with specification by using a calibrated thermometer which will be fitted into a tube containing mineral oil: 120 ºC must be reached inside the tube in a heat block. 7. Carefully remove the heated tubes from the heating block after 6 minutes and place them in the centrifuge. Centrifuge tubes at 10,000 x g for 10 minutes. The supernatant is used for the detection of the galactomannan antigen. 8. Transfer 300ul of supernatant into the second labeled 2ml sample tube which has patient s bar code and will be placed on to the Evolis TM analyzer. 9. After preparation, the supernatant may be removed and stored at 2-8 ºC for up to 48 hours prior to testing. If analysis of the results indicates that retesting of a sample is required, another aliquot of the serum or BAL must be heat treated for retesting. Testing Procedure 1. Bring reagents to room temperature (18-25 ºC) for at least 30 minutes before use. 2. Log onto the system by clicking on OK, no password is required. 3. A self-test of the system is automatically initialized each time the EVOLIS software is run. The self-test is considered satisfactory if the word PASSED appears beside each instrument module. Print a copy of the self-test report and combine it with the work list and result sheets. 4. Down load worklist to EVOLIS Analyzer. 5. Prepare specimens by removing the lids off the heat-treated serum/bal supernatant and load all sample tubes with the barcode facing right on to the sample rack (rack code T). Check the quality of the samples by ensuring all clots, foam and bubbles have been removed.

6 Version: 1.0 CURRENT 6 of Open the door to the sample and reagent unit. Load the sample rack with the bar-codes facing the bar-code reader onto the track marked by the solid red LED light. 7. The Patient Editor dialog box will appear with the specimen numbers from the loaded rack. Double check that the specimen numbers are correct. If there is a blank space under the patient ID column, click on the space and enter the specimen number manually. Select corresponding assay according to the worklist. All specimen numbers entered manually will be flagged with the code ManID on the results report. 8. The Galactomannan assays for BAL and serum should appears along order number Galactomannan assay for BAL= BAL Aspergillus Ag BR V16 Galactomannan assay for serum= Mount Sinai Aspergillus Ag BR V16 9. Change the assay by clicking on the assay box corresponding to the sample number if you want that assay to be run for that sample. A check mark is shown for a sample where the assay has been selected. 10. Once the required assay has been selected for each sample click Close to save. A solid LED light will show up for the next available track in the sample and reagent unit which indicates where the next sample rack can be loaded. Repeat steps 6 to 11 for each new sample rack that is loaded on to the analyzer. 11. A total of four sample racks need to be loaded on to the analyzer even if all the sample racks are not required in order for the reagent template to be used in a later step. When an empty sample rack is loaded on to the analyzer just click Close when the Patient Editor dialog box appears. 12. To check and validate the worklist click on the New Worklist icon located on the upper toolbar and the Set-Up dialog box will appear. 13. Click on + beside the assay file name that is associated with the plate. The plate layout will appear at the right hand side of the dialog box and will show the total number of controls and specimens to be tested, their assigned wells and the number of strips required for the assay. 14. If duplicate samples need to be tested for a given assay in the same run: a) From the Patient Editor dialog box select the assay folder in which the duplicate sample needs to be run and click Add Patient. b) The Select Patient(s) dialog box will appear.

7 Version: 1.0 CURRENT 7 of 23 c) Check the Allow multiple determinations box which appears at the bottom of the Select Patient (s) dialog box. d) The patient sample IDs will appear. Click on the sample ID in which multiple testing is required and click OK. e) In the Set-up Panel dialog box, a (x2) will appear beside the patient ID under the assay folder in which it was selected. 15. If all the information is correct click OK to validate the worklist. 16. A separate dialog box asking for the reagent lot number will appear for each assay that is ordered. Double check the lot number and expiry date and click OK if everything is correct. 17. Click on the + to expand the Work folder and click on Plate Layout, the number of microwell strips required for the assay can be reviewed. Click the Print button located along the top of the toolbar to print the plate layout if desired. 18. Click START (green button) located on the upper tool bar to open the Load dialog box. 19. The Load dialog box appears and shows where to load all the required resources. 20. Load all required resources from left to right: i. Pipette tips: Grey = 1100 ul tips (full rack) Brown= 300 ul tips (full rack) *The pipette tips have to be loaded in the exact position as shown in the layout ii. Load the assay reagents on the reagent racks according to the following templates (Figure 1). Figure 1 is suitable for one batch of heating process Figure 2 is suitable for two batches of heating process The reagent template specific for the Aspergillus Galactomannan assay should be used. iii. Ensure all caps on the reagent/control containers have been removed before loading them onto the analyzer.

8 Version: 1.0 CURRENT 8 of EMPTY 5. Negative Control 6. Cut-Off Control 7. Positive Control Controls for the 1 st batch of heating process Controls for the 2 nd batch of heating process Figure Once all the reagents and controls have been added to the racks, load the reagent racks onto the sample and reagent unit in the following order: 1 st = 0 - Large reagent rack 2 nd = 3 - Small reagent rack 22. In the Load dialog box, click the Open Reagent Layout button. Select the corresponding reagent file and click Open. All reagents and controls will be automatically allocated to their assign position.

9 Version: 1.0 CURRENT 9 of 23 Galactomannan.rea ----one batch of heating process BAL and Blood galactomannan.rea --- two batches of heating process 23. Once all the required resources have been loaded onto the analyzer click OK and the Load dialog box will disappear. The analyzer will begin to check the resources to ensure their amounts are sufficient. 24. The Load Plate dialog box will then appear. Rename the plate ID with the corresponding assay name, for example Galactomannan. The test date will automatically be added to the plate ID by the analyzer. 25. Prepare the required number of microwell strips for the requested assay. The number of strips required is shown in the Plate Layout which is found on the right hand side of the Load Plate dialog box. 26. Insert the microplate into the metal frame microplate holder. Make sure that the A1 position of the plate and holder match. 27. Open the door to the microplate loading compartment and load the microplate and its holder onto the plate transport unit. Once loaded, close the door to the microplate loading compartment. 28. Once the plate is loaded click OK. After the microplate is loaded, the analyzer will start to run automatically. 29. From the Work folder click on Schedule to view the chorological order and completion time of the run. 30. While the samples are being pipetted click on Active Event Log to ensure that all samples were pipetted successfully. Look for any red flags that may appear in the Dispense Sample section which indicates that an error has occurred. The analyzer will pause and flag with an error message if a clot or low sample volume is encountered, allowing for operator intervention. 31. The results report will automatically print out after the assay is completed. 32. Once the assay is finished a dialog box prompting the removable of the test plate and carrier from the analyzer will appear. 33. Open the door to the microplate loading compartment and remove the microplate. After the microplate is unloaded close the door to the microplate loading compartment and click OK.

10 Version: 1.0 CURRENT 10 of A blinking red LED light in the sample and reagent unit indicates that the reagent or sample racks can be unloaded from the analyzer. 35. Do not unload pipette tip racks from the analyzer unless they are completely empty. 36. Close the EVOLIS software. Select File Exit from the menu bar or click the X icon at the top right-hand corner of the Evolis software and shut down the computer. Reporting The presence or absence of galactomannan antigen in the test sample (either serum or BAL) is determined by the calculation of an index for each patient sample. To calculate the Index of a sample, divide the O.D of the patient sample by the mean O.D of the cut-off control. Sample Index = Patient Sample O.D. Mean Cut-off Control O.D Patient serum or BAL with an index < 0.50 is considered to be negative for galactomannan antigen. Patient serum or BAL with an index 0.50 is considered to be positive for galactomannan antigen. All positive specimens should be heat-treated and repeated x2 on the next run. EXCEPTION: A positive 8GAL or 8GALB can be referred, without confirming, for 3weeks after a Confirmed (sample repeated in duplicate) 8GAL or 8GALB. Confirmation must be performed on same sample type, ie. You cannot refer a positive serum result to confirmation results on a BAL. For a sample O.D.= [OD>3.50]: 1. calculate the index for OD=3.50 : 3.50 Mean Cut-off Control O.D 2. Report patient s index > the index of OD=3.50 See Evolis EIA Worksheet for calculations

11 Version: 1.0 CURRENT 11 of 23 All repeats have to be recorder inevolis REPEATS WORKLIST, and give the filled-up worklist to a head LIS officer. An absorbance value of less than may indicate a procedure or instrument error and the result is considered invalid and the specimen should be re-run. Reporting Rules for reporting Index 0.5 Index Initial Reporting After repeat in duplicate 0.5 Index 1.0 Index> Order 8COM 2. Result 8COM:type initial index value 3. Order 8GAL1 and verify 4. Order 8GAL2 and verify 1. 8GAL:Positive Report index in the comment: (Index Values: #.##) *Preliminary testing POSITIVE, to be repeated. 2. Verify 8GAL result 3. Order 8GAL1 and verify 3. Order 8GAL2 and verify 2 of 3 results are positive 8GAL:Positive Report 3 index in the comment: (Index Values: #.##/#.##/#.##) Round off to 2 decimal places 2 of 3 results are Negative 8GAL:Negative Report 3 index in the comment: (Index Values: #.##/#.##/#.##) Round off to 2 decimal places 2 of 3 results are positive 8GAL:Positive Report 3 index in the comment: (Index Values: #.##/#.##/#.##) Round off to 2 decimal places 2 of 3 results are Negative 8GAL:Negative Report 3 index in the comment: (Index Values: #.##/#.##/#.##) Round off to 2 decimal places Trouble shooting No index, but has the following error: P_max_high 1.Order 8COM 2.Result 8COM:type the error Only needs to be repeat once Report according the

12 Version: 1.0 CURRENT 12 of 23 P_min_low P_stop_high P_static_high P_static_high P_mean_low 3.Order 8GAL1 and verify result!

13 Version: 1.0 CURRENT 13 of 23 Maintenance Procedure General Maintenance Tasks Always turn off the instrument before cleaning Daily 1. Perform start-up maintenance (check liquid levels) Make sure the system liquid container is full or almost full and correctly connected to the system. If necessary, refill the liquid-system container. Make sure the liquid waste container is ok. 2. Run the washer maintenance: (Procedure takes approximately 20 min) Fill all wash buffer containers with de-ionized water Click New Worklist located in the upper toolbar The Set-up panel dialog box will appear In the Set-up Panel dialog box click Add Plate In the Set-up Panel dialog box click Add Assay and then select the file WasherClean BR.asy, click Open File. Click OK to validate the various dialog boxes until the worklist appears. Click Start (green button) Load an empty washer microplate with the metal plate holder onto the analyzer when the Load Plate dialog box appears. Click OK after the plate is loaded onto the analyzer. When the run is complete, unload the washer plate and replace the de-ionized water with the appropriate wash buffers. *The daily washer maintenance does not have to be completed on the day the monthly washer maintenance is being done. 3. Perform end-of-day maintenance assay: Check the bag of the tip ejection waste container. If full or nearly full, replace as described. Check the liquid waste level in the liquid waste container, if full or nearly full, decontaminate and empty as described as below: i. Remove cap and pour bleach into the container (without emptying the container first). With ordinary domestic-use bleach (12 chlorine), the

14 Version: 1.0 CURRENT 14 of 23 ii. iii. volume of bleach should equal 10% of the volume of liquid waste in the container. Let it stand for a minimum of 30 minutes (or overnight). Empty the container and rinse it thoroughly with tap water. When adding bleach, it is recommended that you operate under a fume hood as chlorine fumes may occur After adding bleach, do not place the waste container screw cap back on as this may damage the sensor. iv. The screw cap and sensor part may be wiped with a cloth moistened with Microcide (diluted in water at 1.6% 15 ml per 950mL ) Close the EVOLIS software. Select File Exit from the menu bar or click the X icon in the top right-hand corner of the EVOLIS software window. Exit Windows Wait until Windows displays the following message: It is now safe to turn off your computer. Switch off the system (switch on back panel of the instrument). Always exit the EVOLIS software and the Windows software before switching off the system! Open the instrument cover and wipe the tip adapter (pipettor head) using a soft linfree cloth moistened with 70% Ethanol Inspect the instrument (inner and outer surfaces) for stains and spills. If necessary, clean as described as below:

15 Version: 1.0 CURRENT 15 of 23 Outer surfaces Cover and handle Outer panels, including washer window Inner surfaces Dilution and tip rack area Sample and reagent unit Test plate pipetting area Pipettor guide rail Bottom drawer Plate transport unit and compartment Room-temperature incubators Heating incubators Under wash solution containers Computer Computer and monitor Clean with any bactericide, virucide and fungicide hospital-grade disinfectant. To prepare and apply the disinfectant (dilution, spray, wait, wipe ) refer to the instructions prescribed by the product manufacture Same as above Same as above Clean once a month with the same disinfectant as above. Spray the disinfectant on a lint-free paper wipe and wipe the inside of each incubator

16 Version: 1.0 CURRENT 16 of 23 Weekly 1. Run the weekly washer maintenance: (Procedure takes approximately 20 min) Fill all wash buffer containers with de-ionized water Click New Worklist located in the upper toolbar The Set-up panel dialog box will appear In the Set-up Panel dialog box click Add Plate In the Set-up Panel dialog box click Add Assay and then select the file WasherClean BR.asy, click Open File. Click OK to validate the various dialog boxes until the worklist appears. Click Start (green button) Load an empty washer microplate with the metal plate holder onto the analyzer when the Load Plate dialog box appears. Click OK after the plate is loaded onto the analyzer. When the run is complete, unload the washer plate and replace the de-ionized water with the appropriate wash buffers. *The weekly washer maintenance does not have to be completed during the week the monthly washer maintenance is being done. 2. Decontaminate the pipettor wash station: Pour 5 ml of decontamination solution consisting of 1.6% Microcide (15ml of Microcide into 950mL of water) into the pipettor wash station and let it soak for a minimum of 15 minutes. Do not empty, the liquid will drain automatically when the system is initialized. 3. Decontaminate and wipe the tip ejection slide. 4. Clean the instrument surfaces and work area. Monthly 2. Run the monthly washer maintenance: (Procedure takes approximately 1 hour) Fill all the wash buffer containers with de-ionized water Click New Worklist located in the upper toolbar The Set-up panel dialog box will appear In the Set-up Panel dialog box click Add Plate

17 Version: 1.0 CURRENT 17 of 23 In the Set-up Panel dialog box click Add Assay and then select the file WasherManifoldDisinfect BR.asy, click Open File Click OK to validate the various dialog boxes until the worklist appears. Click Start (green button) The Load dialog box will appear. Pour 50ml of 1.6% Microcide into a 60ml of container and load the reagent onto a 0 reagent rack. Then load the rack onto the analyzer. In the Load dialog box, allocate the bottle to the corresponding rack position in which the container was loaded then click OK. Load an empty washer microplate with the metal holder onto the analyzer when the Load Plate dialog box appears. Click OK after the plate is loaded onto the analyzer. A few minutes later a message saying to open the drawer, unscrew waste bottle 1 and close the drawer will appear. Unscrew the cap to waste bottle 1, which is located behind the wash buffer bottles. Close the drawer when done and click OK. After 15 minutes a message prompts you to re-open the drawer and re-screw the cap to waste bottle 1, close the drawer when completed and click OK. When the run is complete, unload the plate and the reagent rack and replace the de-ionized water with the appropriate wash buffers. 3. Decontaminate the sample and reagent racks, plate carrier and tip ejection slide. 4. Decontaminate the system liquid container Empty the system liquid container. Empty the container and rinse thoroughly, twice with tap water and once with de-ionized water. Inspect the filter (attached to the cap) to see if damaged or in need of replacing. Refill the container with freshly prepared System Liquid. Preparation of System Liquid: 10L of de-ionized water. 5. Clean the washer buffer bottles only and NOT the caps & sensor devices. 6. Backup System Files Click Backup button found at the upper tool bar. The System Backup dialog box will open up. Click Backup System Files Once completed, click Close from the System Backup dialog box.

18 Version: 1.0 CURRENT 18 of 23 Quality Control 1. Calculate the mean absorbance of the Cut-Off Control (Cut-Off X): Sum of the O.D values for the two Cut-Off controls divided by 2. The O.D of each Cut-Off control must be and to be considered valid. 2. Calculate the Negative Control Index by dividing the O.D of the Negative Control by the mean absorbance of the Cut-Off Control. Negative Control Index = O.D Negative Control Mean Cut-off Control O.D The index of the Negative Control Serum must be less than 0.40 to be considered valid. 3. Calculate the Positive Control Index by dividing the O.D of the Positive Control by the mean absorbance of the Cut-Off Control. Positive Control Index= O.D Positive Control Mean Cut-off Control O.D The index of the Positive Control Serum must be greater than 2.00 to be considered valid. All QC is checked by senior technologist. Download Worklist To EVOLIS Analyzer 1. Login to the Softlab, double click Instrument Menu under Interface

19 Version: 1.0 CURRENT 19 of High light EVOLIS and click OK button 3. Select Download All in Loadlist drop down menu

20 Version: 1.0 CURRENT 20 of Downloading Procedure window pops up, choose By Order for How to Download 5. For Starting From option, keep the letter and change the first three digits to all 0, then click OK button 6. The Instrument Menu window pops up and shows how many orders will be downloaded. Click OK button

21 Version: 1.0 CURRENT 21 of 23 Related Documents Serology Specimen Management Manual GAL Calculations for Reports Evolis Repeat Worklist Serology External QC Current Serology Inventory Current T:\microbiology\Virology\QC statistics\external QC and INVENTORY Logs\Serology External QC & Inventory CURRENT xls QPEMI03001 Reference: Package insert from Platelia Aspergillus EIA Revised July 2007 EVOLIS Short User Manual software version 1.90, revised June 2008 EVOLIS User Manual software version 1.90, revised June 2000

22 Version: 1.0 CURRENT 22 of 23 Record of Edited Revisions Manual Section Name: Aspergillus Galactomannan Antigen Number / Item Date of Revision Signature of Approval New Heating procedure with template added. QC set done May 25, 2016 Dr. T. Mazzulli with each eating processed. Evolis Repeat Worklist Added to Related documents table June 16, 2016 Dr. T. Mazzulli Reporting table modified: All sites now reporting as UHN. 1 November 22, 2016 Dr. T. Mazzulli General table created. Added note to all reporting round off to 2 decimal places. Annual Review August 8, 2017 Dr. T. Mazzulli Updated rivascope to Microcide cleaning solution and modified quantities. Updated System liquid: removed tween, only 10L of deionized water required. March 21, 2018 Dr. T. Mazzulli

23 Version: 1.0 CURRENT 23 of 23 Number / Item Date of Revision Signature of Approval