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iSED® Automated Erythrocyte Sedimentation Rate (iSED) Analyzer

Correlation Protocol

Scope
Correlation studies (or method comparisons) are performed as part of method validation to assess
whether two analytical methods are comparable. Samples are analyzed using the current method
(reference method) and the new (test) method. Statistical analysis is used to determine agreement.

Sample Requirements1
- Sixty (60) samples with a value distribution between 15 to 105 mm/hr, and at least 40% of the
samples with a pathological state (result over 35 mm/hr)
- Samples from both males and females are to be tested
- Samples must be collected in EDTA anticoagulant 13 x 75 mm closed sample tube (lavender
tube) with a minimum of 3.5mL sample volume
- Samples must be non-hemolyzed and without the presence of clots
- Samples must be processed within four (4) hours from venipuncture
- Hct and MCV data must be collected for each sample to aide in data analysis

Instrument and Materials

- Erythrocyte Sedimentation Rate (ESR) results on the above-mentioned samples run on the
current method
- Complete Blood Count (CBC) results on the above-mentioned samples run on the current
method
- Seditrol® ESR Control Level 1
- Seditrol ESR Control Level 2
- iSED Analyzer

Sample Selection
Choose sixty (60) samples that meet the criteria in the Sample Requirements. The choice of the sample
can be based on the results obtained from processing on the current ESR method.

Running Quality Control Samples on the iSED

1. Press the icon "load sample" on the instrument touch screen and wait for the instrument
mixing wheel to position for loading the sample.
2. Load a Seditrol Level 1 tube with the barcode oriented toward the right for sample ID
recognition.
3. Wait for the sample ID recognition and remove the fingers from the sample loading aperture.

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 4. Wait for the instrument mixing wheel to position to the next available position.

5. Continue to load Seditrol Level 2 tube into the analyzer following steps 2 through 4
6. Wait for the analyzer to complete processing the control samples.
7. Remove the samples from the collection tray.
8. Review results and confirm that the control values are within the acceptable range for each
lot number.

Running Whole Blood Samples on the iSED

1. Press the icon "load sample" on the instrument touch screen and wait for the instrument
mixing wheel to position for loading the sample.

2. The sample wheel rotates to position the next open slot in the sample entry port

3. Insert the tube and sample processing will begin.

4. Repeat steps 1-3 for the next sixty (60) samples.

5. Wait for the analyzer to complete processing the samples.

6. Remove the samples from the collection tray.

7. Save all instrument printouts.

Data Analysis
1. Record the ESR results from the iSED and the current method on the Excel correlation
spreadsheet provided by ALCOR.
2. If available, enter the Hct and MCV values for each sample in the corresponding columns of
the spreadsheet.
3. Attach the spreadsheet to an email message and send it to the Technical Application Specialist
for your region.

REFERENCE
1. Clinical and Laboratory Standards Institute (CLSI). Procedures for the Erythrocyte Sedimentation
Rate Test; Approved Standard—Fifth Edition”. CLSI document H02-A5 (ISBN 1-56238-754-5)

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 Appendix A: Correlation to Manual Westergren Method Protocol

Scope
Correlation studies (or method comparisons) are performed as part of method validation to assess
whether two analytical methods are comparable. Samples are analyzed using the current method and
the new (test) method. If, however there is a discrepancy between the test method and current method
results it may be necessary to compare the test method to a standard reference method. The standard
reference method for the Erythrocyte Sedimentation Rate (ESR) has been determined to be the
Westergren method. Statistical analysis is used to determine agreement.

Sample Requirements
- Samples with a value distribution between 15 to 105 mm/hr, and at least 40% of the samples
with a pathological state (result over 35 mm/hr)
- Samples from both males and females are to be tested
- Samples must be collected in EDTA anticoagulant 13 x 75 mm closed sample tube (lavender
tube) with a minimum of 3.5mL sample volume
- Samples must be non-hemolyzed and without the presence of clots
- Samples must be processed within four (4) hours from venipuncture
- Hct and MCV data must be collected for each sample to aide in data analysis

Instrument and Materials

- Complete Blood Count (CBC) results on the above-mentioned samples run on the current
method
- Seditrol® ESR Control Level 1
- Seditrol ESR Control Level 2
- iSED Analyzer

Sample Selection
Process batches of twenty (20) random samples on the iSED until the desired number of samples has
been collected.

1. Press the “Add sample” icon on the instrument touch screen and wait for the instrument mixing
wheel to position for loading the sample.
2. Load a sample tube with the barcode oriented toward the right for sample ID recognition.
3. Wait for the sample ID recognition and remove the fingers from the sample loading aperture.
4. Wait for the instrument mixing wheel to position to the next available position.
5. Continue to load sample tubes into the analyzer following steps two (2) through four (4) until all
the open positions are filled.
6. Wait for the analyzer to complete processing the samples.
7. Remove the samples from the collection tray.

Running Quality Control Samples on the iSED

1. Press the icon "load sample" on the instrument touch screen and wait for the instrument mixing
wheel to position for loading the sample.

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 2. Load a Seditrol Level 1 tube with the barcode oriented toward the right for sample ID
recognition.
3. Wait for the sample ID recognition and remove the fingers from the sample loading aperture.
4. Wait for the instrument mixing wheel to position to the next available position.

5. Continue to load Seditrol Level 2 tube into the analyzer following steps 2 through 4
6. Wait for the analyzer to complete processing the control samples.
7. Remove the samples from the collection tray.
8. Review results and confirm that the control values are within the acceptable range for each lot
number.

Running Whole Blood Samples using the Westergren Method

1. Process the samples collected from the “Sample Selection” section of this procedure using a
manual Westergren ESR Testing kit. Follow the manufacturer’s instructions when setting up the
test.
2. Some helpful hints to achieve optimized results with the manual method include:
- Assure that you are using fresh samples. Samples should ideally be up to two (2) hours old,
but up to four (4) hours is acceptable
- Do not pour samples from the tube to the ESR cup but pipette a calibrated pipette
- Use a slow pipetting technique to prevent damage to the cells
- Once the samples are transferred to the ESR cups mix the dilution well and very slowly to
make sure the dilution is homogeneous.

Data Analysis
1. Record the ESR results from the iSED analyzer and the current method on the Excel correlation
worksheet provided by ALCOR. To open the spreadsheet, click on the Excel object.

iSED Correlation
Worksheet.xlsx

2. Complete the account information under the “INFO” tab.

3. Click on the “DATA” tab and enter the data for the two (2) methods and, if available, enter the
Hct and MCV values for each sample in the corresponding columns of the spreadsheet.
4. Attach an electronic version of the spreadsheet to an email message and send it to the
Technical Application Specialist for your region.

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