VIDAS® 25 OH Vitamin D TOTAL
Contents
1. le A NEWS NEWS NEWS Medical Ltd 4 4 2013 Vitamin D Vidas MAAN Vitamin D Vidas e 28 e ng ml 8 126 NT o gt 40 VIDAS Vitamin D Total 3 BioM erieux 054 6686183 054 5686303 O go T x 9 0 D 5 7 NNP 10249 49002 Z5 03 9385501 www ilexmedical com sales ilexmedical com 03 9385500 Op9 REF 30 463 ss04004 8 en 201308 VIDAS 25 OH Vitamin D TOTAL VITD VIDAS 25 OH Vitamin D TOTAL VITD is an automated quantitative test for use on the instruments of the VIDAS family for the determination of 25 hydroxyvitamin D Total in human serum or plasma using the ELFA technique Enzyme Linked Fluorescent Assay The VIDAS 25 OH Vitamin D TOTAL assay is to be used a2. At the end of the assay results are automatically calculated by the instrument in relation to the calibration curve stored in memory and then printed out 60 VITD Strips Ready to use Stabilizer of human origin 60 VITD SPRs Ready to use 2 30 Control VITD 1 1 5 mL liquid Calibrator VITD 1 x 2 5 mL liquid Ready to use Interior of SPR coated with vitamin D 25 OH Vitamin D diluted in human serum preservative The concentration in ng mL is indicated on the MLE card after the following mention Calibrator S1 Dose Value The confidence interval in Relative Fluorescence Value RFV is indicated on the MLE card after the following mention Calibrator S1 RFV Range Ready to use 25 OH Vitamin D diluted in human serum preservative The confidence interval in ng mL is indicated on the MLE card after the following mention Control C1 Dose Value Range 1 MLE Card Lot Entry Specifications for the factory master data required to calibrate the test to read the data please refer to the User s Manual 1 Package insert provided in the kit or downloadable from www biomerieux com techlib This product has been tested and shown to be negative for HBs antigen and antibodies to HIV1 HIV2 and HCV However since no existing test method can totally guarantee their absence this product must be treated as potentially infectious Therefore usual safety procedures should be obs
3. natural sera that contain endogenous 25 OH Vitamin D without spiking Samples were analyzed by liquid chromatography coupled to mass spectrometry method LC MS MS in order to determine 25 OH Vitamin D2 and 25 OH Vitamin D3 respective concentrations Samples that were included in this study showed a ratio 25 OH Vitamin D2 25 OH Vitamin D3 gt 4 b 00 np 25 OH D Vidas 25 0H D3 LC MS MS 25 OH Vitamin D2 cross reactivity H H ___ _ x 100 25 OH D2 LC MS MS The mean 25 OH Vitamin D2 cross reactivity for VIDAS 25 OH Vitamin TOTAL assay is 91 Interference The VIDAS 25 OH Vitamin D TOTAL was evaluated for interference consistent with CLSI document EP7 A2 None of the following factors have been found to significantly influence this assay Human Albumin 0 to 60 g L Rheumatoid factors 0 8 577 7 IU mL HAMA 0 8 2 ug mL bioM rieux SA English 6 VIDAS 25 OH VITAMIN D TOTAL VITD 9304004 B en 2013 08 Method comparison 1 A comparison of the VIDAS 25 OH Vitamin D TOTAL assay Y with the IDS iSYS 25 Hydroxy Vitamin D assay X assay gave the following results Number of samples analyzed 344 Equation for Passing Bablok regression Y 0 87 1 Coefficient of correlation 0 93 VIDAS 250H Vitamin D TOTAL vs IDS iSYS 25 Hydroxy Vitamin D Passing amp Bablok e 100 VIDAS 25 OH Vitamin D TOTAL ng mL 0 20 40 60 80 100 IDS iSYS 25 Hydroxy Vitamin ng mL The
4. range are reported as gt 126 0 ng mL Detection and quantitation limits The Limit of Blank is the 95 percentile of more than 60 measurements of analyte free samples LoB corresponds to the concentration below which the probability to obtain analyte free samples is 95 LoB was determined to be 6 2 ng mL The Limit of Detection LoD is the concentration of 25 OH Vitamin D in a sample that can be distinguished from the analyte free sample with a probability of 95 The LoD was determined to be 8 1 ng mL The Limit of Quantitation LoQ is the lowest concentration of 25 OH Vitamin D that can be quantified with a level of acceptable accuracy and precision The LoQ was determined to be 8 1 ng mL The study was performed as recommended by CLSI document EP17 A2 Functional detection limit The functional detection limit is defined as the concentration of 25 OH Vitamin D measured with an inter assay coefficient of variation of 20 During an in house study the functional detection limit was determined to be lt 8 1 ng mL Linearity The VIDAS 25 OH Vitamin D TOTAL assay is linear over its measurement range 7 1 a 126 2 ng mL evaluated according to the recommendations of CLSI document EP6 A Precision A study was performed according to the recommendations of CLSI document EP5 A2 A panel of 5 human samples covering the measurement range was tested as follows each sample was tested in duplicate in 2 separate runs per day over 20 days us
5. S PTC protocol bar codes at the end of the package insert or downloadable from www biomerieux com techlib before reading the MLE data If the MLE data have been read before the VIDAS PTC protocol read the MLE data again Before each new lot of reagents is used specifications or factory master calibration curve data must be entered into the instrument using the MLE data If this operation is not performed before initiating the tests the instrument will not be able to print results The master lot data need only be entered once for each lot It is possible to enter the MLE data manually or automatically depending on the instrument refer to the User s Manual Calibration Calibration using the calibrator provided in the kit must be performed each time a new lot of reagents is opened after the master lot data have been entered Calibration should then be performed every 28 days This operation provides instrument specific calibration curves and compensates for possible minor variations in assay signal throughout the shelf life of the kit English 3 VIDAS 25 OH VITAMIN D TOTAL VITD The calibrator identified by S1 must be tested in duplicate see Users Manual The calibrator value must be within the set RFV Relative Fluorescence Value If this is not the case recalibrate using S1 Procedure 1 Only remove the required reagents from the refrigerator They can be used immediately 2 Use one VITD strip and on
6. ait for samples to coagulate and centrifuge according to the tube manufacturer s recommendations to eliminate fibrin Other tubes follow the tube manufacturers recommendations for use Frozen stored samples after thawing these samples must be homogenized before testing Mix using a vortex type mixer Clarify the samples before analysis by centrifugation if necessary The pre analytical step including the preparation of blood samples is an essential first step when performing medical analyses In conformity with Good Laboratory Practices this step is the responsibility of the laboratory manager Insufficient clot time can result in the formation of fibrin with micro clots that are invisible to the naked eye The presence of fibrin red blood cells or suspended particles can lead to erroneous results Samples containing suspended fibrin particles or erythrocyte stroma should be centrifuged before testing For serum specimens ensure that complete clot formation has taken place prior to centrifugation Some specimens especially those from patients receiving anticoagulant or thrombolytic therapy may exhibit increased clotting times bioM rieux SA 9304004 B en 2013 08 Specimen related interferences Interferences have been studied according to the recommendations of Clinical and Laboratory Standards Institute CLSI document EP7 A2 11 None of the following factors have been found to significantly influence th
7. and Prevention of Vitamin D Deficiency an Endocrine Society Clinical Practice Guideline J Clin Endocrin Metab July 2011 96 7 9304004 B en 2013 08 INDEX OF SYMBOLS 77 T Temperature limit emo Batch code Consult Instructions for Use y Contains sufficient for lt n gt tests WARRANTY bioM rieux disclaims all warranties express or implied including any implied warranties of MERCHANTABILITY AND FITNESS FOR A PARTICULAR USE BioM rieux shall not be liable for any incidental or consequential damages IN NO EVENT SHALL BIOMERIEUX S LIABILITY TO CUSTOMER UNDER ANY CLAIM EXCEED A REFUND OF THE AMOUNT PAID TO BIOMERIEUX FOR THE PRODUCT OR SERVICE WHICH IS THE SUBJECT OF THE CLAIM BIOMERIEUX the blue logo SPR and VIDAS are used pending and or registered trademarks belonging to bioM rieux or one of its subsidiaries or one of its companies CLSI is a trademark belonging to Clinical Laboratory and Standards Institute Inc Any other name or trademark is the property of its respective owner ap bioM rieux SA Z Chemin de l Orme ME RIEU X 69280 Marcy l Etoile France www biomerieux com RCS LYON 673 620 399 Tel 33 0 4 78 87 20 00 Fax 33 0 4 78 87 20 90 CE
8. autoclave solutions containing bleach e The instrument should be regularly cleaned and decontaminated see the User s Manual English 2 VIDAS 25 OH VITAMIN D TOTAL VITD STORAGE CONDITIONS e Store the VIDAS 25 OH Vitamin D TOTAL kit at 2 8 C e Do not freeze reagents e Store all unused reagents at 2 8 C e After opening the kit check that the SPR pouch is correctly sealed and undamaged If not do not use the SPRs e Carefully reseal the pouch with the desiccant inside after use to maintain stability of the SPRs and return the complete kit to 2 8 C e f stored according to the recommended conditions all components are stable until the expiration date indicated on the label SPECIMENS Specimen type and collection Human serum or plasma lithium heparin Do not use EDTA tubes Types of tubes validated Plastic tube with clot activator Plastic tube with separation gel and clot activator Plastic tube with lithium heparin Plastic tube with lithium heparin and separation gel Note Blood collection tube results may vary from one manufacturer to another depending on the materials and additives used It is the responsibility of each laboratory to validate the type of sample tube used and to follow the manufacturer s recommendations for use Samples containing suspended fibrin particles or erythrocyte stroma should be centrifuged before testing Specimen preparation Plain tubes w
9. e VITD SPR from the kit for each sample control or calibrator to be tested Make sure the storage pouch has been carefully resealed after the required SPRs have been removed 3 The test is identified by the VITD code on the instrument The calibrator must be identified by S1 and tested in duplicate If the control is to be tested it should be identified by C1 4 If necessary clarify the samples by centrifugation 5 Mix the calibrator control and samples using a vortex type mixer for serum or plasma separated from the pellet 6 Before pipetting ensure that samples calibrators controls and diluent are free of bubbles For this test the calibrator control and sample test portion is 100 pL 8 Insert the VITD SPRs and VITD strips into the instrument Check to make sure the color labels with the assay code on the SPRs and the Reagent Strips match 9 Initiate the assay as directed in the User s Manual All the assay steps are performed automatically by the instrument 10 Reclose the vials and return them to 2 8 C after pipetting 11 The assay will be completed within approximately 40 minutes After the assay is completed remove the SPRs and strips from the instrument 12 Dispose of the used SPRs and strips into an appropriate recipient RESULTS AND INTERPRETATION Once the assay is completed results are analyzed automatically by the computer Fluorescence is measured twice in the Reagen
10. ed as potentially infectious Therefore usual safety procedures should be observed when handling HARMFUL reagent R20 21 22 Harmful by inhalation in contact with skin and if swallowed R68 20 21 22 Harmful possible risk of irreversible effects through inhalation and in contact with skin 36 37 39 Wear suitable protective clothing gloves and eye face protection IRRITANT reagent R 36 Irritating to eyes S 26 In case of contact with eyes rinse immediately with plenty of water and seek medical advice For further information refer to the Safety Data Sheet available on request MATERIALS AND DISPOSABLES REQUIRED BUT NOT PROVIDED Pipette with disposable tip to dispense 100 uL Powderless disposable gloves For other specific materials and disposables please refer to the Instrument User s Manual Instrument of the VIDAS family WARNINGS AND PRECAUTIONS e For in vitro diagnostic use only e For professional use only e This kit contains products of human origin No known analysis method can totally guarantee the absence of transmissible pathogenic agents It is therefore recommended that these products be treated as potentially infectious and handled observing the usual safety precautions see Laboratory biosafety manual WHO Geneva latest edition This kit contains products of animal origin Certified knowledge of the origin and or sanitary state of the animals does not t
11. erved when handling bioM rieux SA English 1 VIDAS 25 OH VITAMIN D TOTAL VITD The SPR The interior of the SPR is coated during production with vitamin D Each SPR is identified by the VITD code Only remove the required number of SPRs from the pouch and carefully reseal the pouch after opening Description of the VITD strips Sample Well 9304004 8 en 2013 08 The Reagent Strip The strip consists of 10 wells covered with a labeled foil seal The label comprises a bar code which mainly indicates the assay code kit lot number and expiration date The foil of the first well is perforated to facilitate the introduction of the sample The last well of each strip is a cuvette in which the fluorometric reading is performed The wells in the center section of the strip contain the various reagents required for the assay Pre treatment solution TRIS NaCl dissociation agent surfactant methanol Conjugate TRIS NaCl anti vitamin D antibody conjugated with alkaline phosphatase stabilizer of human origin preservative 300 uL 600 pL Reading cuvette with substrate 4 Methyl umbelliferyl phosphate 0 6 mmol l diethanolamine DEA 0 62 mol L or 6 6 pH 9 2 1 g L sodium azide This product has been tested and shown to be negative for HBs antigen and antibodies to HIV1 HIV2 and HCV However since no existing test method can totally guarantee their absence this product must be treat
12. ing two reagent lots 10 test days per reagent lot on 3 instruments N 240 values per sample The 3 instruments were located at one site Two calibrations were used for each reagent lot 5 test days per calibration for each lot The repeatability within run precision and reproducibility within instrument between lot were calculated for each sample using this protocol and are reported in the following table Sample 4 Repeatabilit Within instrument Mean p y between lot reproducibility concentration ng mL Standard Standard deviation deviation ng mL ng mL CV 4 3 2 100 0 bioM rieux SA English 5 VIDAS 25 OH VITAMIN D TOTAL VITD 9304004 B en 2013 08 Specificity The specificity of the VIDAS 25 OH Vitamin D TOTAL assay was assessed by testing cross reactants according to the recommendations of the CLSI document EP7 A2 Cross reactivity was evaluated by adding the following substances to samples containing 25 OH Vitamin D The results of this study are summarized in the following table Tested compound Cross reactivity 1 25 OH 2 Vitamin D2 1 25 OH gt Vitamin D3 81 24 25 OH s Vitamin D3 588 levels tested were 10x to 1000x the typical endogenous levels of analyte Mean Value spiked ng ml Mean Value unspiked ng ml oe od BRS Ee a es Cross reactivity 100 concentration of cross reactant ng ml The cross reactivity P to 25 OH Vitamin D2 was determined using
13. ions in serum or plasma which makes 25 OH vitamin D the preferred analyte for the determination of vitamin D nutritional status 10 KIT COMPOSITION 60 TESTS PRINCIPLE The assay principle combines an enzyme immunoassay competition method with a final fluorescent detection ELFA The Solid Phase Receptacle SPR serves as the solid phase as well as the pipetting device for the assay Reagents for the assay are ready to use and pre dispensed in the sealed reagent strips All of the assay steps are performed automatically by the instrument The reaction medium is cycled in and out of the SPR several times The sample is mixed with pre treatment reagent to separate vitamin D from its binding protein The pre treated sample is then collected and transferred into the well that contains an alkaline phosphatase ALP labeled anti vitamin D antibody conjugate The vitamin D antigen present in the sample and the vitamin D antigen coating the interior of the SPR compete for binding sites on the anti vitamin D antibody ALP conjugate During the final detection step the substrate 4 Methyl umbelliferyl phosphate is cycled in and out of the SPR The conjugate enzyme catalyzes the hydrolysis of this substrate into a fluorescent product 4 Methyl umbelliferone the fluorescence of which is measured at 450 nm The intensity of the fluorescence is inversely proportional to the concentration of vitamin D antigen present in the sample
14. is assay hemolysis after spiking samples with hemoglobin 0 to 1 9 g L monomer lipemia after spiking samples with lipids 0 to 4 0 g L equivalent in triglycerides bilirubinemia after spiking samples with bilirubin 0 to 0 3 g L cholesterol after spiking samples with cholesterol 0 to 5 g L However it is recommended not to use samples that are clearly hemolyzed lipemic or icteric and if possible to collect a new sample Specimen stability Serum and plasma samples can be stored in primary tube at 18 25 C for up to 8 hours or aliquoted at 2 8 C for up to 5 days if longer storage is required freeze the sera or plasma at 25 6 C Serum type samples can be stored for 3 months at 25 6 C with 3 freeze thaw cycles Plasma type samples can be stored for 3 months at 25 6 C with 3 freeze thaw cycles INSTRUCTIONS FOR USE For complete instructions see the Instrument User s Manual VIDAS PTC protocol data entry When using the assay for the first time and before reading the MLE data scan the bar code s at the end of the package insert or downloadable from www biomerieux com techlib using the instrument s external bar code reader This reading will allow VIDAS PTC protocol data to be transferred to the instrument software for its update These data should only be read the first time the assay is used Master lot data entry Note When using the assay for the first time enter the VIDA
15. ning antibodies directed against reagent components For this reason assay results should be interpreted taking into consideration the patient s history and the results of any other tests performed RANGE OF EXPECTED VALUES It is recommended that each laboratory establish its own reference range which may be unique to the population it serves A review of the most recent literature suggests the recommendation for 25 OH Vitamin D levels are 12 Status 25 OH vitamin D A reference range study was conducted using 140 apparently healthy adults based on guidance from CLSI C28 A3 Serum samples were collected in from a French population between January and December and were tested using VIDAS 25 OH Vitamin D TOTAL assay The values observed are summarized below Observed values n 140 25 OH vitamin D ng ml Observed range 2 5th to 97 5th percentile 9 3 48 5 Indicative results the results obtained can vary from one laboratory to another and according to the geographical zones English 4 VIDAS 25 OH VITAMIN D TOTAL VITD 9304004 B en 2013 08 PERFORMANCE Studies performed using VIDAS 25 OH Vitamin D TOTAL kit gave the following results Measurement range The VIDAS 25 OH Vitamin D TOTAL measurement range extends from 8 1 ng mL up to 126 0 ng mL Values below the lower limit of the measurement range are reported as lt 8 1 ng mL Values above the upper limit of the measurement
16. otally guarantee the absence of transmissible pathogenic agents It is therefore recommended that these products be treated as potentially infectious and handled observing the usual safety precautions do not ingest or inhale e Do not use the SPRs if the pouch is pierced or if the dot sealing a SPR is detached bioM rieux SA e Do not use visibly deteriorated STRs damaged foil or plastic e Do not use reagents after the expiration date indicated on the label e Do not mix reagents or disposables from different lots e Use powderless gloves as powder has been reported to cause false results for certain enzyme immunoassay tests e Kit reagents contain sodium azide which can react with lead or copper plumbing to form explosive metal azides If any liquid containing sodium azide is disposed of in the plumbing system drains should be flushed with water to avoid build up e The reading cuvette with pre treatment solution well 3 contains a harmful reagent methanol CH3OH Refer to the risk phrase R and the precautions S above e The reading cuvette with substrate well 10 contains an irritant agent 6 6 diethanolamine Refer to the risk phrase R and the precautions S above e Spills should be wiped up thoroughly after treatment with liquid detergent or a solution of household bleach containing at least 0 5 sodium hypochlorite See the Users Manual for cleaning spills on or in the instrument Do not
17. s an aid in the assessment of Vitamin D sufficiency SUMMARY AND EXPLANATION Vitamin D is a fat soluble steroid prohormone Vitamin D deficiency can be associated with rickets in children and osteoporosis and secondary hyper parathyroidism in adults Recent studies have established a link between low circulating vitamin D levels and an increasing risk of diabetes cardiovascular or autoimmune diseases as well as various forms of cancer 1 8 Vitamin D testing has become an assay of general health status 9 Vitamin D is found mainly in two forms vitamin D2 ergocalciferol and vitamin D3 cholecalciferol Vitamin D3 is synthesized from 7 dehydrocholesterol by action of solar ultraviolet radiation on the skin It is also present in food mostly in fatty fish Vitamin D2 is from exogenous origin only Small amounts of vitamin D2 are present in food mushrooms vegetables Both vitamins D2 and D3 are used for medical supplementation and are identically metabolized by the body The active form of the molecule is the 1 25 OH z vitamin D calcitriol which is obtained from vitamin D through two successive hydroxylation reactions The first hydroxylation occurs in the liver to yield 25 OH vitamin D calcidiol The second hydroxylation occurs in the kidneys and other tissues as well to yield biologically active 1 25 OH 2 vitamin D The 25 OH vitamin D is the main storage form of vitamin D in the human body It is found in high concentrat
18. sample concentrations analyzed with VIDAS 25 OH Vitamin D TOTAL ranged between 8 2 ng mL and 98 2 ng mL 2 A comparison of the VIDAS 25 OH Vitamin D TOTAL assay Y with a LC MS MS method X assay gave the following results Number of samples analyzed 343 Equation for Passing Bablok regression Y 1 00 X 0 41 Coefficient of correlation 0 86 VIDAS 25 OH Vitamin D TOTAL vs LC MS MS 25 OH Vitamin D 120 5 Passing 8 Bablok 100 80 O n 60 Ee gt 5 40 O N lt S 2 gt 0 T T T T T 0 20 40 60 80 100 120 LC MS MS 25 OH Vitamin D ng mL The sample concentrations analyzed with VIDAS 25 OH Vitamin D TOTAL ranged between 8 2 ng mL and 98 2 ng mL bioM rieux SA English 7 VIDAS 25 OH VITAMIN D TOTAL VITD WASTE DISPOSAL Dispose of used or unused reagents as well as any other contaminated disposable materials following procedures for infectious or potentially infectious products It is the responsibility of each laboratory to handle waste and effluents produced according to their nature and degree of hazardousness and to treat and dispose of them or have them treated and disposed of in accordance with any applicable regulations LITERATURE REFERENCES 1 HOLICK M F 2007 Vitamin D deficiency N Engl J Med 357 266 281 2 DOBNIG H et al Independent association of low serum 25 hydroxyvitamin D and 1 25 dihydroxyvitamin D levels wi
19. t Strip s reading cuvette for each sample tested The first reading is a background reading of the substrate cuvette before the SPR is introduced into the substrate The second reading is taken after incubating the substrate with the enzyme remaining on the interior of the SPR The RFV Relative Fluorescence Value is calculated by subtracting the background reading from the final result This calculation appears on the result sheet The results are automatically calculated using calibration curves which are stored by the instrument 4 parameter logistics model and are expressed in ng mL or nmol L Assay results should be used in conjunction with other clinical or laboratory data to assist the clinician in making individual patient management decisions bioM rieux SA 9304004 8 en 2013 08 QUALITY CONTROL One control is included in each VIDAS 25 OH Vitamin D TOTAL kit This control must be performed immediately after opening a new kit to ensure that reagent performance has not been altered Each calibration must also be checked using this control The instrument will only be able to check the control value if it is identified by C1 Results cannot be validated if the control value deviates from the expected value Note It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations LIMITATIONS OF THE METHOD e Interference may be encountered with certain sera contai
20. th all cause and cardiovascular mortality Arch Intern Med 2008 Jun 23 168 12 1340 9 3 FREEDMAN BI et al Vitamin D adiposity and calcified atherosclerotic plaque in african americans J Clin Endocrinol Metab 2010 Mar 95 3 1076 83 4 REIS JP VON MUHLEN D MILLER ER 3rd MICHOS ED APPEL LJ Vitamin D status and cardiometabolic risk factors in the United States adolescent population Pediatrics 2009 Sep 124 3 e371 9 5 ZIPITIS CS AKOBENG AK 5 Vitamin D supplementation in early childhood and risk of type 1 diabetes a systematic review and meta analysis Arch Dis Child 2008 Jun 93 6 512 7 6 PITTAS AG et al 6 Vitamin D and calcium intake in relation to type 2 diabetes in women Diabetes Care 2006 Mar 29 3 650 6 7 YIN L et al Meta analysis longitudinal studies of serum vitamin D and colorectal cancer risk Aliment Pharmacol Ther 2009 Jul 1 30 2 113 25 8 CHEN P etal Meta analysis of vitamin D calcium and the prevention of breast cancer 9 GRANT WB HOLICK MF Benefits and requirements of vitamin D for optimal health a review Altern Med Rev 2005 Jun 10 2 94 111 10 HOLICK MF Vitamin D status measurement interpretation and clinical application Ann Epidemiol 2009 Feb 19 2 73 8 11 Clinical and Laboratory Standards Institute Interference Testing in Clinical Chemistry Approved Guideline Second Edition CLSI document EP7 A2 CLSI 2005 12 HOLICK MF et al Evaluation Treatment
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