Free Estriol ELISA - DIAsource Immunoassays
Contents
1. IMMUNOADSORBENT DIL CAL REC SOLN PEG EXTR SOLN ELU SOLN GEL PRE SOLN NEUTR SOLN TRACEUR BU HRP HRP HRP HRP CONJ BUF CHROM TMB conc CHROM TMB Concentrado de conjugado de biotina Concentrado avidina HRP Tamp n de ensayo Conjugado de biotina Anticuerpo espec fico Estreptavidina HRP Concentrado WASH SOLN Tamp n de lavado Revision nr 1305132. new disposal plastic pipette tips for each calibrator or sample in order to avoid cross contamination Absorbance is a function of the incubation time and temperature Before starting the assay it is recommended that all reagents are ready caps removed all needed wells secured in holder etc This will ensure equal elapsed time for each pipetting step without interruption As a general rule the enzymatic reaction is linearly proportional to time and temperature 6 2 Assay Procedure Each run must include a calibration curve 1 Secure the desired number of Microtiter wells in the frame holder 2 Dispense 10 uL of each Calibrator control and samples with new disposable tips into appropriate wells 3 Dispense 100 uL Enzyme Conjugate into each well Thoroughly mix for 10 seconds It is important to have a complete mixing in this step 4 Incubate for 60 minutes at room temperature 5 Briskly shake out the contents of the wells Rinse the wells 4 times with diluted Wash Solution 400 uL per well Strike the wells sharply on absorbent paper to remove residual droplets Important note The sensitivity and precision of this assay is markedly influenced by the correct performance of the washing procedure Add 100 uL of Substrate Solution to each well Incubate for 30 minutes at room temperature Stop the enzymatic reaction by adding 100 uL of Stop Solution to each well Determine the absorbance OD of each well at 450 10 nm with a microtiter
3. spiked was assayed and analyte concentrations of the samples were calculated from the calibration curve The percentage recoveries were determined by comparing expected and measured values of the samples 2a Sample 2 Sample 3 Average Recovery 100 8 101 8 106 9 Range of Recovery to 103 8 109 8 112 3 9 6 Linearity Sample 3 S S Sample 2 Concentration ng mL 15 58 Average Recovery 100 2 Range of Recovery to 107 9 106 8 103 7 Catalogue Nr KAPD1612 PI number 1701129 en Revision Nr 140127 1 10 LIMITATIONS OF USE Reliable and reproducible results will be obtained when the assay procedure is performed with a complete understanding of the package insert instruction and with adherence to good laboratory practice Any improper handling of samples or modification of this test might influence the results 10 1 Interfering Substances Haemoglobin up to 4 mg mL Bilirubin up to 0 125 mg mL and Triglyceride up to 30 mg mL have no influence on the assay results 10 2 Drug Interferences Until today no substances drugs are known to us which have an influence to the measurement of Free Estriol in a sample 10 3 High Dose Hook Effect No hook effect was observed in this test 11 LEGAL ASPECTS 11 1 Reliability of Results The test must be performed exactly as per the manufacturer s instructions for use Moreover the user must strictly adhere to the rules of GLP Good Laboratory Practice or
4. Calibrador 0 y result ser 0 075 ng mL 9 4 Precisi n Consultar el manual de usuario en ingl s 9 5 Recuperaci n Consultar el manual de usuario en ingl s 9 6 Linealidad Consultar el manual de usuario en ingl s 10 LIMITACIONES DE USO Cualquier manipulaci n inadecuada de las muestras o modificaciones del ensayo pueden influenciar los resultados 10 1 Sustancias que pueden interferir Hemoglobina hasta 4 mg mL Bilirrubina hasta 0 125 mg mL y Triglic ridos hasta 30 mg mL no influencian los resultados del ensayo 10 2 Interferencias con drogas Hasta ahora no se han encontrado sustancias drogas conocidas por nosotros que tengan influencia en la medida de Estriol libre en una muestra 10 3 Efecto Gancho Dosis Elevada No se ha observado efecto gancho en este ensayo 11 ASPECTOS LEGALES 11 1 Fiabilidad de los Resultados El ensayo debe realizarse exactamente de acuerdo a las instrucciones del fabricante Mas a n el usuario debe ajustarse estrictamente a las reglas BPL Buenas Pr cticas de Laboratorio o a otros calibradors y o leyes nacionales aplicables Esto es especialmente relevante para el uso de reactivos control Es importante incluir siempre dentro del procedimiento de ensayo un n mero suficiente de controles para validar la exactitud y la precisi n del ensayo Los resultados del ensayo son v lidos s lo si todos los controles se encuentran dentro de los rangos especificados y si todos los otros par metr
5. DA Ge CE Free Estriol ELISA KAPD1612 DIAsource ImmunoAssays S A Rue du Bosquet 2 B 1348 Louvain la Neuve Belgium LOT 140127 1 FREE ESTRIOL ELISA Enzymeimmunoassay for the quantitative measurement of Free Estriol in human serum KAPD1612 IN VITRO DIAGNOSTIC USE DlAsource ImmunoAssays SA Rue du Bosquet 2 B 1348 Louvain la Neuve Belgium Tel 32 10 84 99 11 Fax 32 10 84 99 90 1 INTRODUCTION 1 1 Intended Use The DlAsource Free Estriol ELISA is an enzyme immunoassay for the quantitative in vitro diagnostic measurement of free estriol unconjugated estriol in serum during the second half of pregnancy This kit is not intended to be used for the risk evaluation of trisomy 21 1 2 Summary and Explanation Estriol E3 is the major estrogen formed by the fetoplacental unit during pregnancy Unconjugated E3 passes through the placenta into the maternal circulation where it is rapidly converted into glucuronide and sulfate derivatives to facilitate its excretion The half life of estriol in the maternal bloodstream is only 20 30 minutes Its measurement therefore offers a convenient and quick evaluation of current fetal status Plasma estriol levels increase steadily throughout pregnancy and most rapidly during the third trimester 28 40 weeks A sudden decrease in fetoplacental E3 production will result in a rapid fall in unconjugated E3 in the maternal serum There are several potential advantages to measuring u
6. alibrator 4 15 0 ng mL Calibrator 5 40 0 ng mL Calibrators 7 EXPECTED VALUES It is strongly recommended that each laboratory should determine its own normal and abnormal values 7 1 Normal healthy adults In a study conducted with apparently normal healthy adults using the DlAsource Free Estriol ELISA the following values are observed Population Valid N 5 Percentile ng mL 95 Percentile ng mL Females not pregnant 0 057 0 822 7 2 Values during pregnancy Week of Expected range Week of Expected range Twin pregnancy gestation gestation p m ng mL p m ng mL ng mL CLINICAL SIGNIFICANCE The measurement of Estriol E3 in body fluids has routinely been used for the monitoring and management of fetal well being particularly in the high risk pregnant patient The concentration of E3 in plasma increases gradually during the first 20 weeks gestation and more rapidly during the third trimester Since the ranges for normal and abnormal serum conjugated E3 are wide and overlap considerably a single E3 determination is of little value The patient should be monitored frequently to establish any individual trend Consistently low levels or a sudden and continual decrease of serum E3 during the third trimester is highly indicative of fetal distress and possibly intrauterine death When such observations are made the status of the fetus should be assessed by alternative methods Interpretation of serum unconjugated E3 levels sho
7. ansporte no est sujeto a la responsabilidad del fabricante 12 REFERENCIAS BIBLIOGRAF A Consultar el manual de usuario en ingl s Fecha de revisi n 2014 01 27 Catalogue Nr KAPD1612 PI number 1701129 es Revision Nr 140127 1 P I Number 1701000 Ag 1251 Ab 1251 Ag 1251 Ab 1251 CONC INC BUF ACETONITRILE SERUM DIL SPE DIL BUF ANTISERUM IMMUNOADSORBENT DIL CAL NEUTR SOLN TRACEUR BUF EXTR SOLN conc WASH SOLN CONTROL N Used symbols Wash buffer Revision nr 130513 P I Number 1701000 S mbolos utilisados Producto sanitario para diagn stico in vitro Fabricante WASH SOLN cone 1251 CONC Ab 1251 conc ACETONITRILE DIL BUF ANTISERUM
8. azards in use and for disposal e Before starting the assay read the instructions completely and carefully Use the valid version of the package insert provided with the kit Be sure that everything is understood The microplate contains snap off strips Unused wells must be stored at 2 C to 8 C in the sealed foil pouch and used in the frame provided e Pipetting of samples and reagents must be done as quickly as possible and in the same sequence for each step e Use reservoirs only for single reagents This especially applies to the substrate reservoirs Using a reservoir for dispensing a substrate solution that had previously been used for the conjugate solution may turn solution coloured Do not pour reagents back into vials as reagent contamination may occur Mix the contents of the microplate wells thoroughly to ensure good test results Do not reuse microwells Do not let wells dry during assay add reagents immediately after completing the rinsing steps Allow the reagents to reach room temperature 21 26 C before starting the test Temperature will affect the absorbance readings of the assay However values for the patient samples will not be affected e Never pipet by mouth and avoid contact of reagents and specimens with skin and mucous membranes e Do not smoke eat drink or apply cosmetics in areas where specimens or kit reagents are handled e Wear disposable latex gloves when handling specimens and reagents Microbia
9. de control de calidad que acompa an al kit Los valores y los rangos fijados en la hoja del control de calidad se refieren siempre al kit actual y deben usarse para la comparaci n directa de los resultados Es recomendable tambi n hacer uso de programas de Aseguramiento de la Calidad nacionales o internacionales para asegurar la exactitud de los resultados Utilizar m todos estad sticos apropiados para el an lisis de los valores y tendencia de los controles Si los resultados del ensayo no se ajustan a los rangos aceptables establecidos en los controles los resultados obtenidos de los pacientes han de considerarse inv lidos En este caso por favor comprobar las siguientes reas t cnicas Pipeteo y tiempo empleado fot metro fecha de caducidad de los reactivos condiciones de almacenamiento e incubaci n m todos de aspiraci n y lavado Catalogue Nr KAPD1612 PI number 1701129 es Revision Nr 140127 1 Despu s de comprobar los asuntos mencionados arriba sin encontrar ning n error contactar con su distribuidor o con DlAsource ImmunoAssays S A directamente 9 CARACTER STICAS DEL ENSAYO 9 1 Rango din mico del ensayo El rango del ensayo se encuentra entre 0 075 40 ng mL 9 2 Especificidad de los Anticuerpos Reactividad Cruzada Consultar el manual de usuario en ingl s 9 3 Sensibilidad Anal tica La sensibilidad anal tica se calcul a partir de la media menos dos desviaciones calibrador de veinte 20 r plicas del
10. de estar a temperatura ambiente antes de su uso Todos los reactivos deben mezclarse sin formar espuma Una vez se ha comenzado el ensayo deben completarse todos los pasos sin interrupci n Utilizar puntas de pipeta de pl stico nuevas para cada est ndar control o muestra para evitar combinaciones cruzadas La absorbancia es funci n del tiempo de incubaci n y la temperatura Antes de comenzar el ensayo se recomienda que todos los reactivos est n preparados tapas removidas todos los pocillos que se necesiten asegurados en recipiente etc Esto asegurar un tiempo similar para cada paso de pipeteo sin que haya interrupciones Como regla general la reacci n enzim tica es linealmente proporcional al tiempo y a la temperatura 6 2 Procedimiento de ensayo Cada uno debe incluir una curva de calibradors Asegurar el n mero deseado de pocillos en el recipiente 2 Dispensar 10 uL de cada Calibrador Control y muestras con puntas nuevas en los pocillos adecuados 3 Dispensar 100 uL de Enzyme Conjugate a cada pocillo Mezclar totalmente durante 10 segundos Es importante mezclar completamente en este paso 4 Incubar durante 60 minutes a temperatura ambiente 5 Sacudir en rgicamente el contenido de los pocillos Lavar los pocillos 4 veces con Wash Solution diluida 400 uL por pocillo Realizar un golpe seco de los pocillos contra el papel absorbente para eliminar las gotas residuales Nota importante La sensibilidad y
11. e ensayo pueden usarse suero No usar muestras hemol ticas ict ricas o lip micas Tener en cuenta No deben usarse muestras que contengan acida s dica 5 1 Toma de muestras Suero Recoger la sangre por punci n en la vena ej Sarstedt Monovette para el suero permitir coagulaci n y separar el suero por centrifugaci n a temperatura ambiente No centrifugar antes de la coagulaci n completa Las muestras de pacientes que reciben terapia anticoagulante requieren m s tiempo para coagular 5 2 Almacenamiento de las muestras Las muestras deben ser tapadas y pueden ser almacenadas hasta 4 d as a 2 C a 8 C antes del ensayo Las muestras almacenadas por un per odo de tiempo mas largo al menos un a o han de congelarse s lo una vez a 20 C antes del ensayo Las muestras descongeladas deben invertirse varias veces antes del ensayo 5 3 Diluci n de las muestras Si en un ensayo inicial se encuentra una muestra que presenta valores mayores que el calibrador mas concentrado ha de diluirse con Calibrador 0 y volver a ensayarse como se describe en el Procedimiento de Ensayo Para el c lculo de las concentraciones habr que tener en cuenta el factor de diluci n Ejemplo a diluci n 1 10 10 uL Suero 90 uL Calibrador O mezclar totalmente b diluci n 1 100 10 uL diluci n a 1 10 90 uL Calibrador 0 mezclar totalmente 6 PROCEDIMIENTO DE ENSAYO 6 1 Consideraciones generales Todos los reactivos y muestras han
12. e used in this assay Do not use haemolytic icteric or lipaemic specimens Please note Samples containing sodium azide should not be used in the assay 5 1 Specimen Collection Serum Collect blood by venipuncture e g Sarstedt Monovette allow to clot and separate serum by centrifugation at room temperature Do not centrifuge before complete clotting has occurred Patients receiving anticoagulant therapy may require increased clotting time 5 2 Specimen Storage Specimens should be capped and may be stored for up to 4 days at 2 8 C prior to assaying Specimens held for a longer time least one year should be frozen only once at 20 C prior to assay Thawed samples should be inverted several times prior to testing 5 3 Specimen Dilution If in an initial assay a specimen is found to contain more than the highest calibrator the specimens can be diluted with Calibrator O and reassayed as described in the Assay Procedure For the calculation of the concentrations this dilution factor has to be taken into account Example a Dilution 1 10 10 uL Serum 90 uL Calibrator O mix thoroughly b Dilution 1 100 10 uL dilution a 1 10 90 uL Calibrator O mix thoroughly 6 TEST PROCEDURE 6 1 General Remarks All reagents and specimens must be allowed to come to room temperature before use All reagents must be mixed without foaming Once the test has been started all steps should be completed without interruption Use
13. es of reagents storage and incubation conditions aspiration and washing methods After checking the above mentioned items without finding any error contact your distributor or DIAsource ImmunoAssays S A directly Catalogue Nr KAPD1612 PI number 1701129 en Revision Nr 140127 1 9 ASSAY CHARACTERISTICS 9 1 Assay Dynamic Range The range of the assay is between 0 075 40 ng mL 9 2 Specificity of Antibodies Cross Reactivity The following substances were tested for cross reactivity of the assay added steroid concentration of steroid OD 450 nm Measured concentration Estriol E3 40 ng ml 39 67 ng ml Testosterone 16 ng ml n d Estradiol E2 2 ng ml n d Estrone E1 2 ng ml n d Cortisol 800 ng ml n d n d non detectable 9 3 Analytical Sensitivity The analytical sensitivity was calculated from the mean minus two standard deviations of twenty 20 replicate analyses of Calibrator O and was found to be 0 075 ng mL 9 4 Precision 9 4 1 Intra Assay Variation The within assay variability is shown below Mean ng mL Cv 9 4 2 Inter Assay Variation The inter assay variability three different kit lots is shown below The between assay variability is shown below Mean ng mL CV 9 5 Recovery Recovery of the DiAsource ELISA was determined by adding increasing amounts of the analyte to three different patient sera containing different amounts of endogenous analyte Each sample non spiked and
14. icuerpo inmovilizado Despu s de la incubaci n el conjugado no unido se lava La cantidad de conjugado de peroxidasa unido es inversamente proporcional a la concentraci n de Estriol en la muestra Despu s de la adici n de la soluci n sustrato la intensidad de color desarrollado es inversamente proporcional a la concentraci n de Estriol libre en la muestra del paciente 3 PRECAUCIONES Este kit es solamente para diagn stico in vitro Por favor se usa solo la version valida de la metodico t cnico incluido aqui en el kit Para obtener informaci n de las sustancias peligrosas incluidas en el kit por favor mirar las hojas de los datos de seguridad del material Todos los reactivos en este kit de ensayo que contienen suero o plasma humano se han ensayado y confirmado ser negativos para HIV I II HBsAg y HCV mediante procedimientos aprobados por la FDA Sin embargo todos los reactivos deben ser tratados tanto en su uso como dispensaci n como potencialmente biopeligrosos Evitar contacto con Stop Solution que contiene H2SO 0 5 M Puede provocar irritaci n y quemaduras en la piel Nunca pipetear con la boca y evitar el contacto de los reactivos y las muestras con la piel y con membranas mucosas No fumar comer beber o usar cosm ticos en reas donde las muestras o los reactivos del kit est n siendo usados Usar guantes de l tex cuando se utilicen las muestras y los reactivos La contaminaci n microbiana de
15. l contamination of reagents or specimens may give false results e Handling should be done in accordance with the procedures defined by an appropriate national biohazard safety guideline or regulation e Do not use reagents beyond expiry date as shown on the kit labels e All indicated volumes have to be performed according to the protocol Optimal test results are only obtained when using calibrated pipettes and microtiter plate readers e Do not mix or use components from kits with different lot numbers It is advised not to exchange wells of different plates even of the same lot The kits may have been shipped or stored under different conditions and the binding characteristics of the plates may result slightly different e Avoid contact with Stop Solution containing 0 5 M H2SO It may cause skin irritation and burns Some reagents contain Proclin 300 BND and or MIT as preservatives In case of contact with eyes or skin flush immediately with water e TMB substrate has an irritant effect on skin and mucosa In case of possible contact wash eyes with an abundant volume of water and skin with soap and abundant water Wash contaminated objects before reusing them If inhaled take the person to open air Catalogue Nr KAPD 1612 PI number 1701129 en Revision Nr 140127 1 Chemicals and prepared or used reagents have to be treated as hazardous waste according to the national biohazard safety guideline or regulation For information o
16. la precisi n de este ensayo se ve marcadamente influenciada por la realizaci n correcta del proceso de lavado 6 Adicionar 100 uL de Substrate Solution a cada pocillo 7 Incubar durante 30 minutes a temperatura ambiente 8 Parar la reacci n enzim tica mediante la adici n de 100 uL de Stop Solution a cada pocillo 9 Leer la OD a 450 10 nm con un lector de microplacas dentro de los 10 minutos despu s de la adici n de la Stop Solution 6 3 C lculo de los Resultados Calcular los valores de absorbancia media para cada conjunto de calibradors controles y muestras de pacientes 2 Construir una curva calibrador mediante la representaci n de la absorbancia media obtenida para cada calibrador frente a su concentraci n con el valor de absorbancia en el eje vertical Y y la concentraci n en el eje horizontal X 3 Usando el valor de absorbancia media de cada muestra determinar la concentraci n correspondiente a partir de la curva est ndar 4 M todo automatizado Los resultados en las instrucciones de uso se han calculado autom ticamente usando una curva de regresi n 4 Par metros 4 Par metros Rodbard o 4 Par metros Marquardt son los m todos preferidos Otras funciones de regresi n dar n lugar a resultados sensiblemente diferentes 5 La concentraci n de las muestra puede leerse directamente de la curva de calibradors Las muestras con concentraciones superiores al mayor calibrador han de diluirse Para el c lculo de las c
17. los reactivos o las muestras puede dar resultados err neos El manejo debe realizarse de acuerdo a los procedimientos definidos por las gu as o regulaci n nacionales de seguridad de sustancias biopeligrosas No utilizar los reactivos despu s de su fecha de caducidad que aparece en las etiquetas del kit Todos los vol menes indicados han de ser realizados de acuerdo con el protocolo Los resultados ptimos del ensayo se obtienen solo cuando se utilizan pipetas y lectores de microplacas calibrados No mezclar o usar componentes de kits con distinto n mero de lote Se recomienda no intercambiar pocillos de distintas placas incluso si son del mismo lote Los kits pueden haber sido enviados o almacenados bajo diferentes condiciones y las caracter sticas de uni n de las placas pueden resultar diferentes Los compuestos qu micos y los reactivos preparados o utilizados han de tratarse como residuos peligrosos de acuerdo con las gu as o regulaci n nacionales de seguridad de sustancias biopeligrosas Las hojas de los datos de seguridad de este producto est n disponibles bajo pedido directamente a DIAsource ImmunoAssays S A Las hojas de los datos de seguridad se ajustan a las demandas de las gu a europea EU Guideline 91 155 EC Catalogue Nr KAPD1612 PI number 1701129 es Revision Nr 140127 1 4 COMPONENTES DEL KIT 4 1 Componentes del Kit 1 Microtiterwells Placas multipocillo 12 x 8 tiras separables 96 pocillo
18. n hazardous substances included in the kit please refer to Material Safety Data Sheets Material Safety Data Sheets for this product are available upon request directly from DlAsource ImmunoAssays S A 4 KIT COMPONENTS 4 1 Contents of the Kit 12x8 break apart strips 96 wells Wells coated with a anti Estriol antibody polyclonal Ze lt N 0 to 5 6 vials 1 mL ready to use See exact values on vial labels Control Low 8 High 2 vials 1 mL each ready to use For control values and ranges please refer to vial label or QC Datasheet Contain non mercury preservative 1 vial 14 mL ready to use Estriol conjugated to horseradish peroxidase Contain non mercury preservative 1 vial 14 mL ready to use Tetramethylbenzidine TMB on gt Q El HEG ies E O fz CHROM TMB 6 1 vial 14 mL ready to use contains 0 5M H2S04 Avoid contact with the stop solution It may cause skin irritations and burns 7 was sorn conc 1 vial 30 mL 40X concentrated wasn sorn conc see Preparation of Reagents A BND 5 bromo 5 nitro 1 3 dioxane MIT 2 methyl 2H isothiazol 3 one Note Additional Calibrator O for sample dilution is available upon request 4 2 Materials required but not provided A microtiter plate calibrated reader 450 10 nm Calibrated variable precision micropipettes Absorbent paper Distilled water Timer Semi logarithmic graph paper or software f
19. nconjugated E3 rather than total serum or urinary E3 Unconjugated estriol levels are free from effects related to maternal renal or hepatic disease and are not altered by the administration of certain antibiotics Unconjugated E3 more accurately reflects fetal outcome in diabetic pregnancies and since no hydrolysis of unconjugated E3 is required a more rapid turnaround for the test result is possible 2 PRINCIPLE OF THE TEST The DlAsource Free Estriol ELISA Kit is a solid phase enzyme linked immunosorbent assay ELISA based on the principle of competitive binding The microtiter wells are coated with a polyclonal rabbit antibody directed towards an antigenic site on the Estriol molecule Endogenous Estriol of a patient sample competes with an Estriol horseradish peroxidase conjugate for binding to the coated antibody After incubation the unbound conjugate is washed off The amount of bound peroxidase conjugate is inversely proportional to the concentration of Estriol in the sample After addition of the substrate solution the intensity of colour developed is inversely proportional to the concentration of Estriol in the patient sample 3 PRECAUTIONS This kit is for in vitro diagnostic use only For professional use only e All reagents of this test kit which contain human serum or plasma have been tested and confirmed negative for HIV III HBsAg and HCV by FDA approved procedures All reagents however should be treated as potential bioh
20. oncentraciones hay que tener en cuenta el factor de diluci n Catalogue Nr KAPD1612 PI number 1701129 es Revision Nr 140127 1 6 3 1 Ejemplo de una Curva Calibrador T pica Los siguientes datos son solamente para la explicaci n y no pueden ser utilizados en lugar de los datos generados en el momento del ensayo Calibrador Unidades pticas 450 nm Calibrador 0 0 0 ng mL Calibrador 1 0 3 ng mL Calibrador 2 1 2 ng mL Calibrador 3 4 0 ng mL Calibrador 4 15 0 ng mL Calibrador 5 40 0 ng mL 7 VALORES ESPERADOS Se recomienda encarecidamente que cada laboratorio determine sus valores normales e inusuales 7 1 Valores con adultos aparentemente sanos En un estudio con adultos aparentemente sanos utilizando el DIAsource Free Estriol ELISA se observaron los siguientes valores 7 2 Valores durante embarazo p m post menstruationem Semanas de Semanas de ra embarazo ng mL er ng mL di ng mL 38 39 7 8 37 13 40 40 42 AR TENER EN CUENTA Para importancia cl nica consultar el manual de usuario en ingl s Cap tulo 7 EXPECTED VALUES 8 CONTROL DE CALIDAD Se recomienda usar muestras control de acuerdo con las leyes estatales y federales El uso de muestras control ser recomienda para asegurar la validez diaria de los resultados Usar controles tanto a niveles normal como patol gico Los controles y los correspondientes resultados del Laboratorio de control de calidad est n fijados en el certificado
21. or data reduction 4 3 Storage and stability of the Kit When stored at 2 8 C unopened reagents will retain reactivity until expiration date Do not use reagents beyond this date Opened reagents must be stored at 2 8 C Microtiter wells must be stored at 2 8 C Once the foil bag has been opened care should be taken to close it tightly again Opened kits retain activity for two months if stored as described above 4 4 Preparation of Reagents Allow all reagents and required number of strips to reach room temperature prior to use Wash Solution Add deionized water to the 40X concentrated Wash Solution Dilute 30 mL of concentrated Wash Solution with 1170 mL deionized water to a final volume of 1200 mL The diluted Wash Solution is stable for 2 weeks at room temperature 4 5 Disposal of the Kit The disposal of the kit must be made according to the national regulations Special information for this product is given in the Material Safety Data Sheets 4 6 Damaged Test Kits In case of any severe damage to the test kit or components DIAsource ImmunoAssays S A has to be informed in writing at the latest one week after receiving the kit Severely damaged single components should not be used for a test run They have to be stored until a final solution has been found After this they should be disposed according to the official regulations Catalogue Nr KAPD1612 PI number 1701129 en Revision Nr 140127 1 5 SPECIMEN Serum should b
22. os del ensayo se encuentran dentro de las especificaciones dadas para el ensayo En cado de alguna duda o inquietud por favor contactar con DlAsource ImmunoAssays S A 11 2 Consecuencias Terap uticas Las consecuencias terap uticas nunca deben basarse s lo en los resultados de laboratorio incluso si todos los resultados del ensayo est n de acuerdo con los asuntos fijados en el punto 11 1 Cualquier resultado de laboratorio es solamente una parte del cuadro cl nico de un paciente Solamente en los casos donde los resultados de laboratorio est n en acuerdo con todo el cuadro cl nico de un paciente se pueden derivar consecuencias terap uticas Nunca deben derivarse consecuencias terap uticas a partir de solamente el resultado obtenido en el ensayo Catalogue Nr KAPD1612 PI number 1701129 es Revision Nr 140127 1 113 Responsabilidad Cualquier modificaci n del kit y o cambio o mezcla de cualquier componente procedentes de kits de lotes diferentes puede afectar negativamente a los resultados esperados y en la validez de todo el test Esas modificaciones y o cambios invalidad cualquier reclamaci n de reposici n Las reclamaciones emitidas debidas a una mala interpretaci n de los resultados de laboratorio por parte del comprador referidos al punto 11 2 son tambi n inv lidas A pesar de todo en el caso de cualquier reclamaci n la responsabilidad del fabricante no excede el valor del kit Cualquier da o provocado al kit durante su tr
23. other applicable national standards and or laws This is especially relevant for the use of control reagents It is important to always include within the test procedure a sufficient number of controls for validating the accuracy and precision of the test The test results are valid only if all controls are within the specified ranges and if all other test parameters are also within the given assay specifications In case of any doubt or concern please contact DiAsource 11 2 Therapeutic Consequences Therapeutic consequences should never be based on laboratory results alone even if all test results are in agreement with the items as stated under point 11 1 Any laboratory result is only a part of the total clinical picture of a patient Only in cases where the laboratory results are in acceptable agreement with the overall clinical picture of the patient should therapeutic consequences be derived The test result itself should never be the sole determinant for deriving any therapeutic consequences 11 3 Liability Any modification of the test kit and or exchange or mixture of any components of different lots from one test kit to another could negatively affect the intended results and validity of the overall test Such modification and or exchanges invalidate any claim for replacement Claims submitted due to customer misinterpretation of laboratory results subject to point 11 2 are also invalid Regardless in the event of any claim the manufact
24. plate reader It is recommended that the wells be read within 10 minutes after adding the Stop Solution e 6 3 Calculation of Results 1 Calculate the average absorbance values for each set of calibrators and patient samples Using semi logarithmic graph paper construct a calibration curve by plotting the mean absorbance obtained from each calibrator against its concentration with absorbance value on the vertical Y axis and concentration on the horizontal X axis 3 Using the mean absorbance value for each sample determine the corresponding concentration from the calibration curve 4 Automated method The results in the IFU have been calculated automatically using a 4 PL 4 Parameter Logistics curve fit 4 Parameter Logistics is the preferred method Other data reduction functions may give slightly different results 5 The concentration of the samples can be read directly from this calibration curve Samples with concentrations higher than that of the highest calibrator have to be further diluted For the calculation of the concentrations this dilution factor has to be taken into account Example of Typical Calibration Curve The following data is for demonstration only and cannot be used in place of data generations at the time of assay Catalogue Nr KAPD1612 PI number 1701129 en Revision Nr 140127 1 Optical Units 450 nm O Calibrator O 0 0 ng mL Calibrator 1 0 3 ng mL Calibrator 2 1 2 ng mL Calibrator 3 4 0 ng mL C
25. rir mantienen su reactividad hasta la fecha de caducidad No utilizar los reactivos mas alla de esta fecha Los reactivos abiertos han de almacenarse a 2 C 8 C Las placas multipocillo han de almacenarse a 2 C 8 C Una vez se ha abierto la bolsa hay que tener cuidado y cerrarla de nuevo Los kits abiertos conservan su actividad durante dos meses si se almacenan como se ha descrito arriba 4 4 Preparacion de los Reactivos Dejar que todos los reactivos y el numero requerido de tiras alcancen la temperatura ambiente antes de usarse Wash Solution Mezclar 30 mL de Wash Solution concentrada con 1170 mL de agua desionizada hasta un volumen final de 1200 mL La solucion del lavado diluida es estable durante 2 semanas a temperatura ambiente 4 5 Eliminacion del Kit La eliminacion del kit debe realizarse de acuerdo con las leyes nacionales En las hojas de datos de seguridad se proporciona informacion especial de este producto 4 6 Kits de ensayo danados En caso de que exista cualquier da o severo del kit de ensayo o de sus componentes ha de informarse por escrito a DlAsource ImmunoAssays S A no mas tarde de una semana despu s de recibir el kit No deben utilizarse componentes da ados para llevar a cabo un ensayo Han de almacenarse hasta que se encuentre una soluci n Despu s de esto deben ser eliminados de acuerdo con las leyes oficiales Catalogue Nr KAPD1612 PI number 1701129 es Revision Nr 140127 1 5 MUESTRAS En est
26. s Pocillos recubiertos con anticuerpo anti Estriol policlonal El r 2 A Calibrador Calibrador 0 5 6 viales 1 mL listo para usar Concentraciones 0 0 3 1 2 4 15 40 ng mL 3 Control Low amp High 2 viales 1 mL listo para usar A Referir los valores y rangos del control a la etiqueta del vial o a la Hoja de datos QC Contiene conservante 4 Enzyme Conjugate Conjugado enzim tico 1 vial 14 mL listo para usar A Estriol conjugado con la Peroxidasa de r bano Contiene conservante sin mercurio Substrate Solution Soluci n de sustrato 1 vial 14 mL listo para usar Tetrametilbencidina TMB Stop Solution Soluci n de parada 1 vial 14 mL listo para usar SOLN contiene 0 5 M H2SO4 Evitar el contacto con la Soluci n de parada Puede causar irritaci n y quemaduras en al piel CHROM TMB STOP Wash Solution Soluci n de lavado 1 vial 30 mL concentrado 40X picota ala ver Preparaci n de los Reactivos BND 5 bromo 5 nitro 1 3 dioxane MIT 2 methyl 2H isothiazol 3 one Nota Se puede solicitar el Calibrador O para la diluci n de la muestra 4 2 Equipamiento y material requerido pero no provisto Lector de microplacas calibrado 450 10 nm ej Microtiter Plate Reader Micropipetas de precisi n variable calibradas Papel absorbente Agua destilada 4 3 Almacenamiento y estabilidad del kit Cuando se almacena a 2 C 8 C los reactivos sin ab
27. uld be made in conjunction with other clinical tests or diagnostic procedures such as amniocentesis and ultrasound Subnormal E3 levels may also be observed in patients being administered certain antibiotics or corticosteroids or in patients with impaired maternal hepatic function 8 QUALITY CONTROL Good laboratory practice requires that controls be run with each calibration curve A statistically significant number of controls should be assayed to establish mean values and acceptable ranges to assure proper performance It is recommended to use control samples according to state and federal regulations The use of control samples is advised to assure the day to day validity of results Use controls at both normal and pathological levels The controls and the corresponding results of the QC Laboratory are stated in the QC certificate added to the kit The values and ranges stated on the QC sheet always refer to the current kit lot and should be used for direct comparison of the results It is also recommended to make use of national or international Quality Assessment programs in order to ensure the accuracy of the results Employ appropriate statistical methods for analysing control values and trends If the results of the assay do not fit to the established acceptable ranges of control materials patient results should be considered invalid In this case please check the following technical areas Pipetting and timing devices photometer expiration dat
28. urer s liability is not to exceed the value of the test kit Any damage caused to the test kit during transportation is not subject to the liability of the manufacturer 12 REFERENCES Bashore R A Westlake J R Plasma unconjugated estriol values in high risk pregnancy Am J Obstet Gynecol June 15 1977 p371 380 Revision date 2014 01 27 Catalogue Nr KAPD1612 PI number 1701129 en Revision Nr 140127 1 FREE ESTRIOL ELISA Enzymeimmunoassay for the quantitative measurement of Free Estriol in human serum KAPD1612 IN VITRO DIAGNOSTIC USE DlAsource ImmunoAssays SA Rue du Bosquet 2 B 1348 Louvain la Neuve Belgium Tel 32 10 84 99 11 Fax 32 10 84 99 90 4 INTRODUCCI N El Kit de inmunoensayo enzim tico DiAsource Free Estriol ELISA proporciona los materiales necesarios para la determinaci n cuantitativa del Estriol libre en suero Este ensayo est dise ado solo para diagn stico in vitro Este kit no esta destinado por ser utilizado para la evaluaci n de riesgo de trisomia 21 2 FUNDAMENTO DEL ENSAYO El Kit DIAsource Free Estriol ELISA es un ensayo en fase s lida de inmunoadsorci n unido a enzimas ELISA basado en el principio de uni n competitiva Los pocillos de las placas est n recubiertos con un anticuerpo policlonal dirigido contra un foci antig nico en la mol cula Estriol En las muestras de los pacientes Estriol compite con un conjugado Estriol peroxidasa de r bano en la uni n al ant
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