Précautions d`utilisation du produit :
Contents
1. The IDS iSYS 1 25D Assay was compared against the IDS 1 25 Dihydroxy Vitamin D AA 54 immunoassay for the quantitative determination of 1 25D following CLSI EP 9A2 Method Comparison and Bias Estimation Using Patient Samples A total of 121 samples selected to represent a wide range of 1 25D concentrations 10 7 197 5 pg mL 25 7 474 0 pmol L was assayed by each method Passing Bablok analysis was performed on the comparative data IDS iSYS 1 00 x IDS RIA 2 8 95 Cl of the slope and intercept were 0 93 to 1 07 and 6 3 to 0 4 respectively correlation coefficient r 0 95 IS 2400PLv02 2012 06 11 English Page 5 5 Specificity Cross Reactivity 1 25 OH 2D2 1 24 25 OH D3 25 OH D3 0 0015 25 OH D2 0 0009 Alfacalcidol 0 04 Bibliography 1 Holick MF Vitamin D deficiency N Engl J Med 2007 357 266 281 2 Shimada T et al FGF 23 is a potent regulator of vitamin D metabolism and phosphate homeostasis J Bone Miner Res 2004 19 429 35 3 K DOQI clinical practice guidelinesfor bone metabolism and disease in chronic kidney disease Am J Kidney Dis 2003 42 Suppl 3 S1 S201 4 Kidney Disease Improving Global Outcomes KDIGO CKD MBD Work Group KDIGO clinical practice guideline for the diagnosis evaluation prevention and treatment of chronic kidney disease mineral and bone disorder CKD MBD Kidney Int 2009 76 Suppl 113 S1 S130 atal Immunodiagnostic Systems Ltd IDS Ltd 10 Di2. uL 200 uL and 1 mL Multi tube vortexer or equivalent End over end or roller mixer Evaporation device or heating block water bath at 40 C and nitrogen supply and manifold Refridgerated centrifuge capable of achieving 2000 g Distilled or deionised water Materials Available on Request Certificate of Analysis Delipidation Procedure 1 Label glass plastic tubes one for each sample Extraction Control Do not delipidate Calibrators Assay Controls 2 Pipette 500 uL of sample Extraction Control to appropriately labelled tubes 3 Add 50 uL of Delipidation Reagent to each tube Vortex all tubes 4 Centrifuge all tubes at 2000 g for 15 minutes NOTE Be careful not to disturb the pellet when handling delipidated samples If the pellet becomes suspended or the sample is not clear repeat the centrifugation Device code IS 2400 i 2 3 4 Instructions for Use Alternative Sample Preparation When the available sample volume is less than 500 uL IS 2400 Label conical bottom plastic tubes or micro tubes one for each sample Add sample e g 300 uL to appropriately labelled tubes Add 0 1 x sample volume of Delipidation Reagent e g 30 uL to each tube Vortex all tubes Centrifuge all tubes at 2000 g for 15 minutes Immunoextraction Procedure l Label Immunocapsules for each sample extraction control Do not immunopurify Calibrators Assay Controls NOTE 10 11 DO NOT USE if
3. IS 2400 IDS iISYS 1 25 Dihydroxy Vitamin D immuno Instructions for Use IN VITRO DIAGNOSTIC CE Intended Use For In Vitro Diagnostic Use The IDS iSYS 1 25 Dihydroxy Vitamin D kit is a complete assay system intended for the purification of 1 25 dihydroxyvitamin D 1 25D in human serum or plasma by immunopurification followed by quantitative determination on the IDS iSYS Multi Discipline Automated System System to assess 1 25 dihydroxyvitamin D deficiency associated with renal disease Results are to be used in conjunction with other clinical and laboratory data to assist the clinician in making patient management decisions Summary and Explanation There are two forms of vitamin D vitamin D3 and vitamin Do Vitamin Ds cholecalciferol is the naturally occurring vitamin D that is produced in the skin after exposure of 7 dehydrocholesterol to solar ultraviolet radiation 1 Vitamin D is manufactured through the ultraviolet irradiation of ergosterol from yeast Both are used in vitamin D supplements The vitamin D compound is biologically inactive but enters the circulation and is hydroxylated in the liver to 25 hydroxyvitamin D 25D which is used to determine a patient s vitamin D status In the kidney 25D is further hydroxylated to produce biologically active metabolite 1 25 dihydroxyvitamin D 1 25D 1 1 25D is one of the major regulators of calcium and phosphate metabolism stimulating intestinal calcium absor
4. an Immunocapsule shows signs of leakage or incorrect volume Vortex Immunocapsules Allow solid phase to settle Stand Immunocapsules upright in provided cardboard inlay for 3 5 minutes Remove Immunocapsules screw caps Add 150 uL of delipidated sample extraction control to Immunocapsules Replace caps securely Place Immunocapsules in cardboard inlay and rotate end over end at 5 20 revolutions per minute for 90 minutes at room temperature 18 25 C Stand Immunocapsules upright for 3 5 minutes allowing gel to settle a Tap to dislodge any gel adhering to the screw caps b Allow gel to settle for a further 1 2 minutes Remove screw cap break off do not twist off bottom stopper from Immunocapsules a Place each Immunocapsule in a plastic glass tube b Centrifuge at low speed 500 1000 g for approximately 1 minute Add 500 uL of distilled or deionised water to each Immunocapsule Add carefully to avoid solid phase splashing out of the Immunocapsule a Centrifuge at low speed 500 1000 g for approximately 1 minute b Repeat above step for further two 2 times for a total of 3 wash cycles Label 2 mL polypropylene conical skirted base screw cap micro tubes one for each Immunocapsule Transfer Immunocapsules to the appropriated labelled micro tubes Add 150 uL of Elution Reagent to all Immunocapsules Allow reagent to soak for 1 to 2 minutes a Centrifuge at low speed 500 1000 g for approximately 1 minu
5. be found on the mini CD Use calibrator levels A and B to adjust the master curve to the reagents on board the System Check for the presence of a 1 25D cartridge on the reagent tray and the availability of the cartridge master curve in the database If the data for the lot of calibrators is not available on board the system load the data using the mini CD provided with the calibrator The calibration is carried out in triplicate RLU CVs of gt 7 will result in a failed calibration One replicate may be removed to meet the calibration requirements The Assay Controls must also be measured in duplicate to calibrate the system Verify and approve the calibration according to the calibration status displayed in the calibration windows and discard the calibrator after use Calibration The IDS iSYS 1 25D assay has been standardized against in house reference standards Calibration Frequency A new calibration is required e Each time a new lot of cartridge is loaded on board Each time a new lot of trigger or cuvette is used e When the control values do not fall within the defined ranges e When the calibration interval of 21 days has expired e After System service Verification of the calibration is automatic and managed by the IDS iSYS system Quality Control The IDS iSYS 1 25 Dihydroxy Vitamin D Control Set IS 2430 is required for quality control The Assay Controls should be measured prior to loading the im
6. ble Range The reportable range of the assay is 6 5 210 0 pg mL 15 6 504 pmol L Any value that reads below 6 5 pg mL 15 6 pmol L should be reported as lt 6 5 pg mL lt 15 6 pmol L The highest reportable value without dilution is 210 0 pg mL 504 pmol L Limitations of Use 1 As in the case of any diagnostic procedure results must be interpreted in conjunction with the patients clinical presentation and other information available to the physician 2 The performance characteristics of this assay have not been established in a paediatric population 3 The following substances do not interfere in the IDS iSYS 1 25D Assay when the concentrations presented in the following table are below the stated threshold Potentially Interfering Threshold Expected Values Each laboratory should determine ranges for their local population The following range was determined using the IDS iSYS 1 25D Assay and is provided for information only The 95 reference interval for the following group was calculated by a non parametric method following the NCCLS guideline C28 A2 How to Define and Determine Reference Intervals in the Clinical Laboratory 26 1 95 0 pg mL n 119 62 6 228 pmol L n 119 Normal Adults Device code IS 2400 IS 2400 he g h IDS iSYS 1 25 Dihydroxy Vitamin D immuno IN VITRO DIAGNOSTIC Instructions for Use Performance Data Representative pe
7. dcot Way Boldon Business Park Boldon Tyne amp Wear NE35 9PD England Tel 44 191 519 0660 Fax 44 191 519 0760 e mail info ukK idsplc com www idsplc com Immunodiagnostic Systems UK Immunodiagnostic Systems Ltd IDS Ltd 10 Didcot Way Boldon Business Park Boldon Tyne amp Wear NE35 9PD England Tel 44 191 519 0660 Fax 44 191 519 0760 e mail info ukK idsplc com www idsplc com USA Immunodiagnostic Systems Inc 8425 N 90th Street Suite 8 Scottsdale AZ 85258 Tel 1 480 278 8333 Fax 1 480 836 7437 e mail info us idsplc com www idsplic com Germany Immunodiagnostic Systems GmbH IDS GmbH Mainzer Landstrasse 49 60329 Frankfurt am Main Tel 49 69 3085 5025 Fax 49 69 3085 5125 e mail info de idsplc com www idsplc com France Immunodiagnostic Systems IDS 153 Avenue D Italie 75013 Paris France Tel 0 1 40 77 04 50 Fax 0 1 40 77 04 55 e mail info fr idsplc com www idsplc com Scandinavia Immunodiagnostic Systems Nordic a s IDS Nordic a s Marielundvej 30 2 Sal 2730 Herlev Danmark Tel 45 44 84 0091 e mail info nordic idsplc com www idsplc com Belgium Immunodiagnostic Systems S A Rue E Solvay 101 4000 Li ge Belgium Tel 32 4 252 26 36 Fax 32 4 252 51 96 e mail info be idsplc com www idsplc com Device code IS 2400
8. dge Magnetic particles coated with streptavidin in a phosphate buffer containing bovine serum albumin and sodium azide as preservative lt 0 1 1 vial 2 6 mL per vial Conjugate 1 25D labelled with an acridinium ester derivative in a phosphate buffer containing bovine serum albumin with sodium azide as preservative lt 0 1 1 cartridge vial 9 6 mL per vial Antibody Biotin Anti 1 25D polyclonal antibody labelled with biotin in a phosphate buffer containing sheep proteins with 0 01 sodium azide as preservative 1 cartridge vial 13mL per vial BUFD Wash buffer 1 proprietary detergent in PBS azide 1 cartridge vial 35mL per vial Calibrators CAL A CAL B Lyophilised MOPS buffer containing bovine serum albumin 1 25D and sodium azide as preservative lt 1 0 2 each of 2 concentration levels 1 2mL Mini CD Contains IFU for IDS iSYS 1 25D reagents and CRY files Materials Required But Not Provided IDS iSYS Multi Discipline Automated System IS 310400 IDS iSYS 1 25 Dihydroxy Vitamin D Control Set IS 2430 IDS iSYS Cuvettes Cube IS CC100 IDS iSYS System Liquid IS CS100 IDS iSYS Wash Solution IS CW 100 IDS iSYS Triggers Set A and B IS CT100 IDS iSYS Cartridge Check System IS 6010 Disposable 12 x 75 mm tubes Disposable polypropylene 2 mL 10 8 mm diameter conical skirted base screw cap micro tubes and screw cap with O ring Sarstedt 72 664 and 65 716 or equivalent Precision pipettes to deliver 150
9. econstitute Invert calibrators gently before use Do not vortex the calibrators Pipette approximately 500 uL 120 uL per replicate of calibrators into 2 mL polypropylene conical skirted base micro tubes and place on the instrument within 15 minutes Device code IS 2400 IS 2400 IDS iISYS 1 25 Dihydroxy Vitamin D immuno Instructions for Use IN VITRO DIAGNOSTIC CE of reconstitution Proceed according to the instructions of the IDS iSYS User Manual If calibrators are to be used more than once they should be stored at 20 C or lower within 15 minutes of reconstitution When re using frozen calibrator vials thaw at room temperature and mix well Ensure that calibrators are at room temperature before pipetting approximately 500 uL of calibrators into 2 mL polypropylene conical skirted based micro tubes and place on the instrument Calibrators should be placed on the system within 15 minutes of reaching room temperature NOTE i DISCARD the material in the micro tubes after use ii DO NOT return material to the calibrator vials System Calibration Two 1 25D calibrators are required to perform the adjustment of the master curve The calibrators are supplied with the kit calibrators from another kit lot must not be used The IDS iSYS 1 25D Assay Controls IS 2430 MUST be measured at the same time as the calibrators to perform a master curve adjustment All data required for the calibration of the cartridge batch can
10. is inversely proportional to the amount of 1 25D present in the samples Warnings and Precautions The IDS iSYS 1 25 Dihydroxy Vitamin D kit is for in vitro diagnostic use only and is not for internal use in humans or animals This product must be used strictly in accordance with the instructions set out in these Instructions for Use IFU IDS Limited will not be held responsible for any loss or damage except as required by statute howsoever caused arising out of non compliance with the instructions provided CAUTION This kit contains material of animal origin Handle kit reagents as if capable of transmitting an infectious agent Appropriate precautions and good laboratory practice must be used in the storage handling and disposal of the kit reagents Disposal of kit reagents should be in accordance with local regulations Sodium Azide Xn Harmful Calibrators contain sodium azide NaN3 gt 0 1 w w lt 1 R22 Wear suitable protective clothing and gloves R52 53 Harmful to aquatic organisms may cause long term adverse effects in the aquatic environment S46 If swallowed seek medical advice immediately and show this container or label S36 37 Wear suitable protective clothing and gloves S60 This material and or its container must be disposed of as hazardous waste Some reagents in this kit contain sodium azide which may react with lead copper or brass plumbing to form highly explosive metal azides On disposal flu
11. munopurified samples extraction controls onto the system to ensure the integrity of the system and the cartridge To verify the validity of sample results the Extraction Controls should be immunopurified and measured at the same time as the patient samples Extraction Controls should be tested at the beginning of every run containing patient samples or according to local regulations Refer to the IDS iSYS 1 25D Control Set IS 2430 Instructions for Use for preparation and handling procedures IS 2400PLv02 2012 06 11 English Page 4 5 Determination of Sample 1 25D levels Proceed according to the instructions of the IDS iSYS User Manual for assaying samples Calculation of Results The 1 25D concentration of each sample is calculated automatically The display of the concentrations screen or printed is produced per user setting To convert results to SI units pmol L pg mL x 2 4 The IDS iSYS 1 25D Assay uses a 4 parameter logistic curve fit 4PL to calculate the 1 25D concentrations Validation of Samples Results Verify the immunopurified Extraction Controls values against the acceptable limits specified in the Control Certificate If an immunopurified Extraction Control value is outside the acceptable limits the samples results are invalid In this case repeat the delipidation and immunopurification for the samples Extraction Controls prior to the determination of 1 25D levels procedure Measurement Range Reporta
12. ollection and Storage The assay should be performed using serum standard sampling tubes or tubes containing serum separating gel or plasma lithium heparin sodium heparin or potassium EDTA samples Samples should be separated as soon as possible after collection Samples Storage Stability Duration Room temperature 24 Hours 20 C or lower 7 Days 2 5 Months To minimise possible evaporation effects reconstituted calibrators should be measured within 3 hours and reconstituted immunopurified samples within 90 minutes after being placed on the system It is recommended to follow the instructions of the tube manufacturer especially when processing samples in primary tubes NOTE Some sample collection tubes that are commercially available might affect the results of testing in particular cases Freeze thaw cycles Procedure Materials Provided Immunoextraction Kit SORB Immunocapsules containing monoclonal antibody to 1 25D linked to solid phase particles in suspension with vitamin D binding protein inhibitor 100 immunocapsules REAG 1 Delipidation Reagent a solution of dextran sulphate and magnesium chloride 1 bottle 6 mL per bottle REAG 2 Elution Reagent Ethanol 2 bottles 25 mL per bottle BUF Assay buffer a MOPS buffer containing bovine serum albumin with 0 01 sodium azide 1 bottle 22 mL per bottle IS 2400PLv02 2012 06 11 English Page 2 5 IN VITRO DIAGNOSTIC Reagent Cartri
13. ption and increasing bone resorption It also inhibits parathyroid hormone PTH production both by direct action on the parathyroid glands and indirectly by raising serum calcium levels 1 25D production is itself stimulated by parathyroid hormone PTH thus providing an effective control loop 1 Fibroblast growth factor 23 FGF 23 secreted from the bone causes the sodium phosphate co transporter to be internalized by the cells of the kidney and small intestine and also suppresses 1 25D synthesis 2 In secondary hyperparathyroidism a disease outside of the parathyroids causes all of the parathyroid glands to become enlarged and hyperactive It is usually caused by kidney failure a problem where the kidney is unable to clean the blood of phosphorus produced by the body and unable to make enough vitamin D specifically 1 25D the active form of vitamin D The build up of phosphorous leads to low levels of calcium in the blood which stimulates the parathyroid glands to increase parathyroid hormone PTH production leading to the glands to enlarge As the disease progresses the parathyroid glands no longer respond normally to calcium and Vitamin D The rationale for direct activated vitamin D therapy in CKD is to slow the progression of secondary hyperparathyroidism The clinical practice guidelines such as the Kidney Disease Outcomes Quality Initiatives KDOQ and the Kidney Disease Improving Global Outcomes KDIGO recommend activa
14. rformance data are shown Results obtained at individual laboratories may vary Sensitivity The limit of blank LoB limit of detection LoD and limit of quantitation LoQ were determined with guidance from CLSI EP17 A Protocols for Determination of Limits of Detection and Limits of Quantitation using 100 blanks and 50 low level samples 28 pgml 6 7 pmol 6 5 pg mL 15 6 pmol L 12 2 pgimL 29 3 pmol L Precision Precision was evaluated in accordance with a modified protocol based on CLSI EP 5A2 Evaluation of Precision Performance of Quantitative Measurement Methods Three serum controls were assayed using three lots of reagents in duplicate twice per day for 20 days on three instruments Concentration EREA CACARE eo ms 7a Concentration pmol L 153 8 370 3 Linearity Linearity was evaluated based on CLSI EP 6A Evaluation of the Linearity of Quantitative Measurement Procedures A Statistical Approach Samples containing varying concentrations of 1 25D were assayed in duplicate The resulting mean concentrations were compared to predicted concentrations Samples were prepared by diluting a high patient sample with a low patient sample prior to extraction 12 sets of samples covering the range of the assay were measured giving a total of 108 measured dilutions Average Observed Expected 1 25D values were 98 4 with an r squared of 0 98 defined by linear regression analysis Method Comparison
15. sh with large volumes of water to prevent azide build up Elution Reagent Elution reagent contains ethanol R11 Highly flammable flashpoint 13 C S7 Keep container tightly closed S16 Keep away from sources of ignition No smoking Handling Precautions Apart from the calibrators which are lyophilised the reagents provided in the kit are ready to use Refer to the calibrator section of the procedure for reconstitution methodology Before a new cartridge is loaded onto the IDS iSYS system mix the magnetic particles container by a brisk rotation motion This will resuspend the magnetic particles that have settled during shipment Ensure that there is no foam formation in the cartridge reagents Should this occur store the cartridge in an upright position in the dark at 2 to 8 C until foaming has dissipated Device code IS 2400 IS 2400 IDS iISYS 1 25 Dihydroxy Vitamin D immuno Instructions for Use Shelf Life and Storage of Reagents Prior to first use store the cartridge and the calibrators in an upright position in the dark at 2 to 8 C Do not freeze the cartridge Reagent shelf life Cartridge Calibrators Before opening at 2 8 C To the expiry date Cartridge After opening at2 8 C ices da a On board the IDS iSYS 49 Days Calibrators After reconstitution at 2 8 C we Calibrators After reconstitution at 20 N A 28 Days C or lower Continuous on board stability Calibrators Sample C
16. te to collect eluate b Repeat above step a further two times The total elution volume collected is 450 uL for each sample extraction control Discard Immunocapsules Place micro tubes ina heating block or water bath set to 40 C Evaporate the eluates under a gentle flow approximately 2 4 psi of nitrogen gas Avoid splashing of the eluates Evaporation should take approximately 45 minutes NOTE IS 2400PLv02 2012 06 11 English Eluants must be completely dry IDS iISYS 1 25 Dihydroxy Vitamin D Page 3 5 immuno IN VITRO DIAGNOSTIC Alternative Evaporation Devices Other evaporation equipments such as vacuum centrifuge evaporator are feasible for use for Step 11 Laboratory should validate the procedure to ensure the optimal setting of their equipments 12 Add 200 uL of Assay Buffer to each tube Vortex for at least 5 seconds to dissolve residues The immunopurified samples extraction controls are now ready for assay 13 Load the immunopurified samples Extraction Controls onto the IDS iSYS System NOTE Cap the immunopurified samples Extraction Controls if they are not intended to be loaded onto and measured by the IDS iSYS system within 90 minutes to minimise their evaporation Immunopurified Samples Storage and Stability The immunopurified samples must be loaded onto and measured by the IDS iSYS System within 90 minutes after reconstitution Otherwise cap the immunopurified samples micro tubes s
17. ted vitamin D therapeutic regimens for CKD patients 3 4 Method Description The IDS iSYS 1 25 Dihydroxy Vitamin D kit is a complete assay system for the purification of 1 25D in patient samples by immunopurification followed by quantitation by one site chemiluminescent immunoassay on the automated IDS iSYS instrument 150yuL delipidated sample is added to an immunocapsule which contains a gel containing a monoclonal anti 1 25D antibody 90 minutes rotation of the immunocapsule allows the binding of 1 25D to the monoclonal antibody The gel is washed to remove potential interfering substances and the 1 25D eluted with ethanol Eluates are then evaporated under IS 2400PLv02 2012 06 11 English Page 1 5 a gentle flow of nitrogen at 40 C and reconstituted in 200 uL assay buffer The reconstituted immunopurified samples are placed into the IDS iSYS sample rack the sample rack is then loaded on the IDS iSYS system 120 uL of the reconstituted immunopurified samples are incubated with the biotinylated sheep anti 1 25D antibody The 1 25D Acridinium conjugate is then added which competes for antibody binding sites Streptavidin coated magnetic particles are then added and following a further incubation step the particles are washed to remove unbound materials Following the addition of Trigger Reagents a flash chemiluminescent reaction is initiated The light signal is measured by the photomultiplier as Relative Light Units RLU and
18. tore in an upright position Ensure the immunopurified samples are at room temperature before loading onto the system Immunopurified Before On board the IDS iSYS N A 90 Minutes Continuous on board stability Assay Procedure IDS iSYS Multi Discipline Automated System Settings The IDS iSYS system s automatic validation of results feature should be disabled Reagent Cartridge The reagents provided in the cartridge are ready to use The system automatically performs the mixing of magnetic particles to maintain homogeneity Before a new cartridge is loaded on board the system mix the magnetic particles container by brisk rotation motion Ensure that there is no foam formation in the cartridge reagents Should this occur store the cartridge in an upright position in the dark at 2 to 8 C until foaming has dissipated The barcode is read when the cartridge is loaded on the reagent tray If the label cannot be read by the system barcode reader a manual procedure exists to enter the barcode data see the IDS iSYS User Manual Load the cartridge on the reagent tray and wait for at least 40 minutes before starting the assay If the cartridge is removed from the reagent tray store the cartridge vertically at 2 8 C in the dark Calibrators The 1 25D calibrators are lyophilised Reconstitute immediately before use Add 1 2 mL of distilled or deionised water to each bottle Replace the stopper Leave for 10 minutes to r
Download Pdf Manuals
Related Search