The following protocol is used to evaluate the ESR measurement method based on the ESR measurement reference method mentioned in NCCLS Booklet H2-A5 (CLSI). After the installation and commissioning of the device in the laboratory and the training of the relevant user by the support company, the quality evaluation steps are carried out in the following order. Checking the specifications of the ESR device A: Birth Certificate Specifications: The device must have identification details including the name and model of the device, the name of the manufacturing company, the name of the manufacturer country, the manufacturing serial number, electrical information and warnings that the device is able to identify and announce. b) Carrying instructions for use and maintenance C) Announcing the type and volume of the sample required for anticoagulant d) Carrying special tubes for collecting samples. E) Having memory to store patients’ results And (having a rotator for the pipes of the machine) Checking the performance of the machine Before starting the investigation process, it is necessary to ensure the suitability of the pipes of the device (in terms of size, volume and stability of the pipes in the place of the pipes in the machine) and the appropriate anticoagulant (sodium citrate 3.2%) and the accuracy of the anticoagulation ratio in the pipes and the accuracy of sampling. Very Important Points – The test should be performed at a temperature of 18-25 °C and the sample is stable at this temperature for a maximum of 4 hours after sampling. – The size of the sediment pipes should be 120 x 8 mm (8 mm diameter of pipe and 120 mm length of pipe). – If the sample is stored in the refrigerator (temperature of 4 degrees Celsius) for up to 12 hours after the sampling, it is possible to do There are experiments. The sample temperature should be reached at room temperature before performing the test. – The tube containing the citrate sample should be scrutinized at least 12 times before the test. – Glass tubes should be used because of their high transparency as well as a suitable surface tension with red blood cells. – Because after the first sedimentation, the molecular attraction of red blood cells decreases and the first sedimentation cannot be expected for the second time, therefore, for a more accurate comparison, two samples must be taken from each patient so that one can be used for the device and the other for the manual method, and at the same time, the tests will be started. – For a deviation of 3 degrees of pipette, the final solution of the nebula increases by 30%. – The vibration created on the test table during the test increases the sodium. – Increasing the temperature by increasing the Rolex increases the sodium. – The deformation of the cells such as spherocytosis and sickle-like prevents the creation of a Rolex and slows down the velocity. – Hepartin increases the rate of sedimentation by changing the zeta potential and causing wrinkles in the blood cells. Imprecision Checking In order to control the repeatability, the inaccuracy of the device should be calculated. For this purpose, when taking blood from a person, a sample is prepared with 10 special sediment tubes and these samples are taken by The device is tested and their results are recorded. With 10 results, the mean of SD standard deviation and standard deviation coefficient This CV is the inaccuracy of a series of tasks called the Within Run CV. This should be done for at least 10 people. with ESR in normal and abnormal ranges. Calculate SD (Standard Deviation) and CV based on the following formula. Article Parsian Teb Zaman Company Sediman ESR Machine and Manufacturer of Python Full Automatic Elize If the CV% obtained is less than or equivalent to the laboratory inaccuracy allowed for ESR testing or the manufacturer’s claim stated in the device instructions, the device reproducibility is considered appropriate and the inaccuracy check is performed, the maximum acceptable level of CV=% inaccuracy is suggested. Inaccuracy Device Inaccuracy Control Reference Method Explanation: In the reference method, the sample should be prepared with anticoagulant K2EDTA.dH2O or Na2EDTA.2H2O or K3EDTA with a concentration of 1.4-2 mg/ml, and the PCV (Packed Cell) volume of the samples should be less than 0.35. Samples should be tested using the Westergren method with standard pipettes. Standard Pipette Pipettes for sodium testing should: – They should be colorless and circular. -200 mm long and divided into 1 mm spacing from 0 to 200. – The diameter of the piping strips should not be less than 2.55 mm. – They should be free of any cracks and fractures and do not have any leakage. In this method, at least 40 samples with 15-105 mm ESR and 5 PCV (Packed Cell Volume) less than 0.35 mm will be selected. If the PCV of a sample is higher than 0.35, it can be corrected in the following way: First, the sample is divided into two separate tubes equally and one of them is centrifuged to separate the plasma. If the volume of the divided samples in each tube is 3.5 ml, the amount of plasma required in milliliters, which must be added to the uncentrifuged tube to adjust its hematocrit less than or equal to 0.35, is calculated using the following formula. Article Parsian Teb Zaman Company Manufacturer Fully Automatic Python Ilizer Conducting the Experiment In this step, 40 subjects with ESR between 15-105 mm are selected and three sets of ESR samples are collected from each person. 1. One on the special tubes of the device that contain 3.2% citrate for measurement with the device. 2. A series on EDTA without preparation of dilution with citrate for the reference method. 3. A series with sodium citrate anticoagulant for the usual Westergrain method (manual method) Tips: Before performing sedimentation test, by measuring the hematocrit of the samples, it should be ensured that the hematocrit is adjusted to a value less than or equal to 0.35. To perform the test, the first series of samples is tested with a device and the second series is tested by the reference method. In the reference method, the test is performed at a temperature of 18-25 °C and the pipettes are placed in a vertical position and without vibration. Results are read at 1±60 times. The results obtained from the instrumental method and the reference method are compared using the following table. For example, if the sodium of a sample in the reference method was 20, according to the table, the number obtained from the machine method should be between 5-17. In this method, if 95% of the answers are within the specified range, the results are acceptable. The maximum acceptable level of bias inaccuracy = 5% is suggested. Article of Parsian Teb Zaman Company Hematology Mixer Machine, Sodiman, Manufacturer of Python Full Automatic Elez Article of Parsian Teb Zaman Company Hematology Mixer Machine, Sodiman, Manufacturer of Python Full Automatic Elez Method selected by NCCLS Selected Method At this stage, based on the NCCLS suggestion, at least 40 samples with ESR in normal and abnormal ranges should be tested with the usual Westergreen method and the results should be compared with the instrumental method. To perform this method, we can use the third series samples that are described in the reference method section. The consistency of the results of the Westergreen method with the instrumental method should be calculated by Paired Result T-test, which is also available in SPSS statistical software. The permissible amount of T is determined based on the number of tested samples and using the T-table available in the statistical books. For example, for 10 samples, the T number should be less than 2.26. Author:Dr. Parisa Dahim Fariba Sabzavi