S TEST Reagent Cartridge Low Density Lipoprotein Cholesterol (LDL)
Contents
1. up to 25 mg dL Lipemia no interference up to 614 mg dL Ascorbic acid no interference up to 50 mg dL Lack of interference was defined as recoveries between 90 and 110 of the neat value and assay performance claims were established on the HITACHI Clinical Analyzer by testing two serum pools containing approximately 30 mg dL and 60 mg dL LDL Cholesterol Precision per CLSI EP5 A2 Four levels of serum samples were assayed 2 times per run 2 runs per day for total of 20 days The precision was found to be Mean mg dL _SD mg dL 3 1 6o 34 n 80 per level Precision POL sites Three levels of samples A B and C were tested by three POL sites six times a day for five days The precision estimates are described below T mg dl aes or a f e f os fie f os fis 4 eo os eo e o o eleo a s m s i iz iis e o es 21 e2 20 e f e ee p e f a CX n 30 replicates per sample per site Oj Patient Correlation POL sites A series of 50 or more serum specimens with LDL Cholesterol values ranging from 25 mg dL to 283 mg dL were assayed on the HITACHI Clinical Analyzer at three sites using S TEST LDL y and a comparative method as the reference method x Linear regression analysis least squares yielded the following results Site a Range Regression ap SE CI Si imewen mg dL Equation mg dL Slope p 26 to 274 y 0 94x 3 8 0 99 8 1 0 90 to 0 99 1 7 to 9 4 a 512512. used for any other purposes Storage and Stability LDL reagent cartridges are stable until the expiration date shown on the box labels when stored in the refrigerator at 2 8 C Specimen Requirements Patient Preparation No special patient preparation is required Collect specimen by standard laboratory technique Specimen Collection 1 Use clear unhemolyzed serum or heparin plasma 2 Care should be taken to preserve the chemical integrity of the blood specimen from the time it is collected until the time it is assayed see SPECIMEN HANDLING AND STORAGE Specimen Identification Label each specimen tube with the patient s identification name and or number Specimen Handling and Storage Before the measurement the sample must be brought back to room temperature 15 30 C If collected serum plasma cannot be tested on the same day store it as follows e For testing within 1 week 2 10 C e For testing after 1 week or longer below 20 C Test Procedure For complete information on operation see the User Manual for the HITACHI Clinical Analyzer Equipment Required HITACHI Clinical Analyzer Reagent Required S TEST LDL reagent cartridge Material Required but not provided 1 Two levels of controls 2 Sample cups 3 Disposable transfer pipettes 4 Washing water 5 Alkali detergent 6 Waste container Assay Procedure Prior to performing each run check system status to determine the need to
3. 0276 y 0 93x 64 038 80 08910098 0610123 CI 95 confidence interval Patient Correlation laboratory site A series of 122 serum specimens with LDL Cholesterol values ranging from 8 mg dL to 370 mg dL were assayed on the HITACHI Clinical Analyzer using S TEST LDL y and a comparative method as the reference method x Linear regression analysis least squares yielded the following results i Range Regression ap SE CI Si nieicegi mg dL Equation mg dL Slope P 8 to 370 y 0 94x 7 6 0 90 to 0 97 3 0 to 12 1 CI 95 confidence interval Serum Plasma Comparison Study A study was performed to validate the use of heparinized plasma as well as serum for the HITACHI Clinical Analyzer with S TEST LDL Approximately 40 matched serum plasma samples that spanned the dynamic range were assayed in singleton and the results were compared using least squares liner regression plasma y axis The performance characteristics were as follows y 1 01x 4 4 correlation coefficient r 0 99 95 confidence interval of the slope 0 99 to 1 02 95 confidence interval of the y intercept 6 4 to 2 3 Detection limit per CLSI EP17 A The detection limit was determined to be 0 8 mg dL Linearity per CLSI EP6 A The assay linearity was determined to be 3 mg dL to 430 mg dL Reportable Range 8 mg dL to 400 mg dL Routine Maintenance and Troubleshooting For complete information on operation see the User Manual for th
4. HitachiChemical vo S TEST Reagent Cartridge Low Density Lipoprotein Cholesterol LDL 96313 10 10 S TEST Cartridges 96313 100 10 x 10 S TEST Boxes 100 Tests Kit for the determination of Low Density Lipoprotein Cholesterol on HITACHI Clinical Analyzer 2011 11 Rev 0 Intended Use The S TEST LDL is intended for the quantitative determination of low density lipoprotein cholesterol concentration in serum or heparin plasma using the HITACHI Clinical Analyzer The S TEST LDL is intended for use in clinical laboratories or physician office laboratories For in vitro diagnostic use only Method Enzymatic direct method Test Summary and Explanation Low Density Lipoprotein LDL Cholesterol is known to be a risk factor for arteriosclerotic disease especially coronary disease Determination of total cholesterol has been conventionally and widely used in the diagnosis of hyperlipidemia which is considered a cause of arteriosclerotic disease However it has been reported that ischemic heart disease is more strongly correlated with LDL Cholesterol than with total cholesterol LDL Cholesterol level is generally calculated from the measurements of total cholesterol HDL Cholesterol and triglycerides using Friedewald s formula but this calculation method is not sufficient for an accurate determination of LDL Cholesterol The reference method for LDL Cholesterol is the ultracentrifugation method but the method requires specialized instru
5. e HITACHI Clinical Analyzer Technical Support Instrument Service 1 First contact to your local distributor 2 Hitachi Chemical Co Ltd Japan Reference 1 Japan Atherosclerosis Society Guideline Investigating Committee for Diagnosis and Treatment of Hyperlipidemia Journal of Atherosclerosis and Thrombosis 25 1 34 1997 2 Japanese Journal of Clinical Medicine 62 Special Issue 18 21 2004 3 Third Report of National Cholesterol Education Program NCEP Expert Panel on Detection Evaluation and Treatment of High Cholesterol in Adults Adult Treatment Panel Ill Executive Summary 2002 4 CLSI Document EP5 A2 Evaluation of Precision Performance of Quantitative Measurement Methods Approved Guideline 5 CLSI Document EP6 A Evaluation of Linearity of Quantitative Measurement Procedures Approved Guideline 6 CLSI Document EP7 A2 Interference Testing in Clinical Chemistry Approved Guideline 7 CLSI Document EP17 A Protocols for the Determination of Limits of Detection and Limits of Quantitation Approved Guideline 80 7868 00 00 05 12 Hitachi Chemical Co Ltd pel 2 1 1 Nishi Shinjuku Shinjuku ku Tokyo 163 0449 Japan
6. ery 30 days and 1 When test results do not match patient symptoms or clinical findings 2 When using a new lot or shipment of reagents 3 When laboratory environmental conditions have significantly changed 4 When training or retraining of personnel occurs Reading and Reporting Results Expected Value e Reportable range 8 400 mg dL e Reference range 70 139 mg dL 2 e Optimal lt 100 mg dL Near optimal 100 129 mg dL Borderline high 130 159 mg dL High 160 189 mg dL Very high gt 189 mg dL 3 e It is recommended that each laboratory determine the expected values for its particular population Interpretation of Results There may be reactions with non target substances or interfering reactions If measured results seem unreliable repeat the measurement if necessary after dilution or try another analytical measurement Handling Critical Values If the result of a sample exceeds the measurement range dilute the sample with physiological saline solution and repeat the measurement Performance Characteristics Please note this assay has not been certified by the Cholesterol Reference Method Laboratory Network CRMLN but is traceable to the CRMLN method Interference per CLSI EP7 A2 The data demonstrated that the LDL test system was not affected by high levels of the following substances at the levels noted Hemoglobin no interference up to 1000 mg dL Unconjugated bilirubin no interference
7. l lt 2 5 U mL e Cholesterol oxidase Microbial 1 2 U mL e Peroxidase Horseradish lt 1 3 ppg U mL e Surfactant 1 e Good s Buffer pH 6 3 LDL Reagent 2 e Surfactant 2 e N N bis 4 sulfobutyl m toluidine disodium salt DSBmT 0 04 e Good s Buffer pH 6 3 Preparation and Labeling LDL reagent is provided in a ready to use cartridge The 2D code label on the front of each cartridge automatically identifies the reagent to the system Reagent Cartridge Reagent 1 Reagent 2 2D code Photometric cuvette Reagent cells Reaction cell Precautions 1 This product should be stored to avoid freezing Frozen reagent should not be used Reagent exceeding the expiration date should not be used Avoid direct sunlight during storage and measurement Avoid all contact with cartridge contents do not ingest When handling samples use disposable gloves to avoid the danger of infection Do not pipette by mouth LDL Reagent 1 and 2 contain Proclin an antiseptic which may be irritating to eyes skin and mucous membranes or BR WP O Disposal Precautions 1 When handling blood and used cartridges use disposable gloves to avoid the danger of infection 2 The samples and reagent cartridges should be disposed of as medical waste in accordance with local regulations Other Precautions This product is intended for use on HITACHI Clinical Analyzer The reagent cartridges should not be
8. mentation and a long measurement time This makes the reference method difficult to perform in routine laboratory tests and direct methods are widely used Principle of the Test The presented method using a combination of two surfactants is based on a fact that each lipoprotein reacts with different surfactants depending on their intrinsic physicochemical property In the first reaction Surfactant 1 changes the structure of only those lipoproteins other than LDL i e chylomicron CM VLDL and HDL and the resulting cholesterol is consumed by cholesterol oxidase and cholesterol esterase in a colorless reaction In the second reaction the remaining LDL is changed by Surfactant 2 and the resulting cholesterol is consumed in a color reaction The first reaction Surfactant 1 HDL VLDL CM Cholesterol Cholesterol esterase Cholesterol _ _______ H202 Cholesterol oxidase i Peroxidase H202 4 Aminoantipyrine _ gt Colorless The second reaction Surfactant 2 LDL __YXYX Y Cholesterol Cholesterol esterase Cholesterol YY H202 Cholesterol oxidase a Peroxidase H202 4 Aminoantipyrine DSBmT Purple red pigment Reagent Requirements one cartridge per patient sample Reagent Composition The S TEST LDL reagent cartridge has the following composition LDL Reagent 1 e 4 Aminoantipyrine 0 01 e Cholesterol esterase Microbia
9. replace washing water or empty waste container See the User Manual for detailed operating instructions Measurement Measurement 546 660 nm 546 660 nm Reagent 1 Sample Reagent 2 v Conc Calculation 210 uL 5 pL 70 uL 0 45 5 0 10 0 min Presentation of Result Each Patient Report includes the data and time sample ID number as programmed the test abbreviation the test results normal ranges and result flags For detailed explanations on flags and error messages refer to the User Manual Calibration Each lot of S TEST LDL cartridges is calibrated by the manufacturer prior to shipment using material traceable to the Beta Quantification Reference Method CDC The 2D code printed on each cartridge provides the analyzer with lot specific calibration data Calculation LDL Cholesterol concentration is directly determined by multiplying the change in absorbance of the unknown samples by the calibrator factor on the 2D code Patient and control results appear on the display Quality Control Users should follow federal state and local regulatory requirements regarding quality control practices See instrument manual for procedures on how to run controls Good laboratory practice includes the use of at least two levels of control material to ensure the test performance The frequency and limits of QC testing should be determined according to individual laboratory standard QC procedures Controls should be run at least once ev
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