CELL-DYN ImmunoPlt (CD61)

Transcription

1 CELL-DYN ImmunoPlt (CD61) REF REAGENT 03H tests 1. NAME CELL-DYN ImmunoPlt (CD61) Reagent 2. INTENDED USE CELL-DYN ImmunoPlt (CD61) Reagent is a one-color direct immunofluorescence reagent for enumerating platelets (PLT) in human whole blood, when used in combination with the CELL-DYN Sapphire System or the CELL-DYN 4000 System (CELL-DYN 4000 software version 8.2 or greater). The ImmunoPlt (CD61) Reagent is for in vitro diagnostic use only. 3. SUMMARY AND EXPLANATION OF THE TEST Human PLT (thrombocytes) are small anucleated (non-nucleated) cellular structures released from the cytoplasm of megakaryocytes. enables automated enumeration of CD61 labeled PLT. Research studies indicate that CD61 PLT analyses provide improved PLT counting in thrombocytopenic patients. 1,2 The blood of many thrombocytopenic patients contains significant numbers of non-platelet particles which resemble platelets both in size and some physical properties such as electrical impedance and light scatter. The non-platelet particles have been shown to include erythrocyte fragments, white cell fragments, immune complexes, bacteria and perhaps others. They are seen especially in patients with thrombocytopenia accompanied by hemolytic anemia, immune complex formation due to underlying autoimmune disease, or drug reaction...second, accurate platelet counts have become of steadily increasing importance because of the pressure to decrease the use of platelet transfusions and to improve the inventory control for platelets in the blood bank BIOLOGICAL PRINCIPLES OF THE PROCEDURE The CD61 antigen is found on all normal resting and activated human PLT. 3 The CD61 antibody is directed against gpiiia of the gpiib/ IIIa complex. 4 In the CELL-DYN ImmunoPlt (CD61) Reagent, fluorescein isothiocyanate (FITC) is conjugated to a CD61 monoclonal antibody (mab). The instrument aspirates a sample from an EDTA-anticoagulated human whole blood specimen, mixes it with the ImmunoPlt (CD61) Reagent and dilutes the mixture for processing. The solid-state laser in the CELL-DYN Sapphire System or the argon-ion laser in the CELL-DYN 4000 System excites the FITC-labeled PLT with 488 nm laser light. The resulting light scatter and fluorescence are measured at 90 o, 7 o, and FL1 fluorescence (530 nm) respectively. This information is processed by specific algorithms to produce the ImmunoPlt (CD61) count. 5. REAGENT Reagent Provided, Sufficient for 40 Tests The CELL-DYN ImmunoPlt (CD61) Reagent is packaged in two pouches. Each pouch contains 20 tubes. Each tube contains sufficient lyophilized reagent for one test. Each tube contains a lyophilized bead of FITC-labeled CD61 antibody in a protective matrix. The fluorescein-to-protein ratio (F: P) for the CELL-DYN ImmunoPlt (CD61) Reagent is 2.8 to 4.5. The F:P ratio has been optimized for its intended use. Specificity: The CELL-DYN ImmunoPlt (CD61) Reagent recognizes an Mr 110-kDalton (kda) protein, also known as gpiiia, the common β-subunit (integrin β 3 -chain) of the gpiib/iiia complex and the vitronectin receptor (VNR). 3,4 Clone: The CELL-DYN ImmunoPlt (CD61) Reagent, clone RUU-PL 7F12, 5 is derived from hybridization of mouse P3X63. Ag8.653 myeloma cells with spleen cells from BALB/c mice immunized with purified PLT membrane glycoproteins. 5 Ig Chain Composition: The antibody is composed of mouse IgG 1 heavy chains and kappa light chains. 5 CELL-DYN ImmunoPlt (CD61) Reagent Page 1 of 6

2 Warnings and Precautions IVD In Vitro Diagnostic Medical Device. Not for irrigation, infusion or ingestion. CAUTION: This product requires the handling of human specimens. It is recommended that all human sourced materials are considered potentially infectious and be handled in accordance with the OSHA Standard on Bloodborne Pathogens 6. Biosafety Level 2 7 or other appropriate biosafety practices 8,9 should be used for materials that contain or are suspected of containing infectious agents. Information for European customers: For product not classified as dangerous per European Directive 1999/45/EC Safety data sheet available for professional user on request. Storage Instructions When pouches are stored unopened and at room temperature, the antibody reagent is stable until the expiration date shown on the label. Do not use after the expiration date. Protect tubes from moisture and ambient light. Always reseal the foil pouch immediately after opening it. Once the pouch has been opened and resealed, use the remaining tubes within 60 days. 30 Once a tube has been removed from a pouch, it 15 is stable for 8 hours at temperatures between 15 o and 30 o C. When blood and Diluent/Sheath Reagent are added to the CELL-DYN ImmunoPlt (CD61) Reagent, the dilution is usable for one test only; the reagent tube should then be discarded into an appropriate biohazard waste container. Indications of deterioration The indicator granules in the desiccant package should be a blue color. If the indicator granules have turned to lavendar color, do not use the product. Alteration in the appearance of the reagent pellet, such as a change in color (from pink to red), or shrinking of the pellet (so that it can pass through the stainless steel retainer when the tube is inverted), indicates instability and deterioration. In such cases, the reagent tube should be discarded. 6. INSTRUMENT The CELL-DYN ImmunoPlt (CD61) Reagent has been validated for use on the CELL-DYN Sapphire and the CELL-DYN 4000 Systems. For detailed information concerning specific ImmunoPlt (CD61) Assay instructions, refer to the CELL-DYN System Operator s Manual, Section: ImmunoPlt (CD61) Assay. 7. SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS Use only human whole blood specimens, anticoagulated with EDTA. Specimens can be stored at room temperature or refrigerated (2 to 8 C) and run within 36 hours of collection. If refrigerated specimens are used, allow them to warm to room temperature before use. 8. PROCEDURE CELL-DYN ImmunoPlt (CD61) Reagent is designed for use on the CELL-DYN Sapphire and CELL-DYN 4000 Hematology Systems which includes all required reagents, with appropriate hardware and software. NOTE: The CELL-DYN 4000 must be equipped with software version 8.2 or greater. All performance characteristics were obtained using a CELL-DYN Sapphire and a CELL-DYN 4000 System. For detailed information, refer to the CELL-DYN System Operator s Manual, Section: ImmunoPlt (CD61) Assay. automatically processes a human whole blood sample with a proprietary, lyophilized FITC-labeled CD61 monoclonal antibody reagent. The method uses a solid-state laser or an argon-ion laser for light scatter and CELL-DYN ImmunoPlt (CD61) Reagent Page 2 of 6

3 fluorescence detection of PLT tagged with the FITC-labeled CD61 antibody at 530 nm. automatically configures the optics, adjusts the electronics and analyzes the data. When the blood and Diluent/Sheath Reagent are added to the ImmunoPlt (CD61) Reagent, automated processing begins. Each ImmunoPlt (CD61) Reagent tube may be used only once and should then be discarded. Performance Characteristics and Specifications Refer to Section 12 in this Package Insert. Operating Instructions Refer to the CELL-DYN System Operator s Manual, Section: ImmunoPlt (CD61) Assay. Operational Precautions and Limitations For detailed information, refer to the CELL-DYN System s Operator Manual, Section: ImmunoPlt (CD61) Assay. Reagent Background Count Procedure For background counts, a tube of CELL-DYN ImmunoPlt (CD61) Reagent is run with an empty specimen tube to check for particulate material in the reagent and system. It is recommended that a background count be run with each new lot of CELL-DYN ImmunoPlt (CD61) Reagent and also with each new lot of Diluent/ Sheath Reagent. Calibration The CELL-DYN Immuno-Plt count is factory calibrated. Quality Control Procedures It is recommended that a whole blood specimen from an adult with PLT values in the normal range be used for the patient controls. This specimen should be undiluted as a normal control and diluted 1:4 with Diluent/Sheath reagent as a low control. The two levels of control should be run once per day of operation and results confirmed to be within acceptable limits before reporting any patient results. Patient controls should be handled according to the laboratory s protocol. The patient control s ImmunoPlt (CD61) count is compared to the patient control s optical PLT count in an automated, internal QC program. The acceptable limit for the ratio of the ImmunoPlt (CD61) count to the optical PLT count is for the normal control and for the low control. The optical PLT count is calibrated and controlled with CELL-DYN HemCal Plus and CELL-DYN 29 Plus (with Retic) Control respectively. For detailed System Calibration and QC information, refer to the CELL-DYN System Operator s Manual, Sections: 6 Calibration and 11 Quality Control. For detailed ImmunoPlt (CD61) QC information, refer to the CELL-DYN System Operator s Manual, Section : ImmunoPlt (CD61) Assay. 9. RESULTS CELL-DYN ImmunoPlt (CD61) results are acquired using information from 90 o and 7 o scatter and fluorescence (FL1) channels. This method differentiates PLT from red cells, white cells, cellular debris and other particles using light scatter and the fluorescence of the FITC-CD61 monoclonal antibody, bound to CD61 antigens on the surface of the PLT. The software applies algorithms to the measured data and produces final results, automatically; no calculations by the operator are required. Flagging Refer to the CELL-DYN Systems Operator s Manual, Section: ImmunoPlt (CD61) Assay for information and descriptions of result flags. CELL-DYN ImmunoPlt (CD61) Reagent Page 3 of 6

4 10. LIMITATIONS OF THE PROCEDURE requires the presence of CD61 antigen and an accurate red cell analysis. If interfering substances are present in sufficient numbers, they may interfere with the algorithms used to obtain the ImmunoPlt (CD61) count. The following list of interfering substances and conditions can affect the results of the CELL-DYN ImmunoPlt (CD61) Assay and is neither all-inclusive or non-exclusive: Glanzmann s thrombasthenia PLT aggregation Platelet satellitism RBC auto-fluorescence Cold or autoagglutinins Human anti-mouse anti-bodies NOTE: Any substance or condition that can interfere with the System s principles of operation should be considered an interfering substance or condition. All potential interferents have not been formally tested by Abbott. 11. EXPECTED VALUES Each laboratory should establish its own reference intervals for this assay. 12. SPECIFIC PERFORMANCE CHARACTERISTICS For details on the methods used to establish performance specifications, refer to the CELL-DYN Sapphire or the CELL-DYN 4000 Systems Operator s Manual, Section: CELL-DYN ImmunoPlt (CD61) Assay. Refer to the CELL-DYN Sapphire or the CELL-DYN 4000 Systems Operator s Manual, Section: CELL-DYN ImmunoPlt (CD61) Assay for instrument specific system performance results for: CD61 Background (Count) Reproducibility (Precision) Analytical Measurement Range (Linearity) Comparability (Correlation) Carryover 13. BIBLIOGRAPHY 1. Ault KA, et al. Implementation of the immunological platelet count on a hematology analyzer the Abbott CELL-DYN Lab Hematol. 1997; 3: Dickerhoff R, Von Rueker. Enumeration of platelets by multiparameter flow cytometry using platelet-specific antibodies and fluorescent reference particles. Clin Lab Haem. 1995; 17: von dem Borne AEGKr, et al. Platelet antibodies, the overall results. In: Knapp W, et al, eds. Leukocyte Typing IV, White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1989: Modderman PW. Cluster report: CD61. In: Knapp W, Dorken B, Gilks WR, et al, eds. Leukocyte Typing IV, White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1989: Monoclonal Antibodies Detecting Human Antigens; CD61. In: Becton Dickinson Immunocytometry Systems Monoclonal Antibodies Source Book. 1996: US Department of Labor, Occupational Safety and Health Administration. 29 CFR Part Occupational Exposure to Bloodborne Pathogens. 7. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories, Fourth Edition. Washington, D.C.: US Government Printing Office. May World Health Organization. Laboratory Biosafety Manual. Geneva: World Health Organization, National Committee for Clinical Laboratory Standards. Protection of Laboratory Workers from Occupationally Acquired Infections: Approved Guideline Second Edition. NCCLS Document M29-A2. Wayne, PA: NCCLS, CELL-DYN ImmunoPlt (CD61) Reagent Page 4 of 6

5 KEY TO SYMBOLS USED 14. NAME AND PLACE OF BUSINESS OF MANUFACTURER EC REP Authorized Representative in the European Community i CAUTION Consult instructions for use Manufactured for: Abbott Laboratories Abbott Park, IL USA 2010 IVD In Vitro Diagnostic Medical Device REAGENT Reagent EC REP ABBOTT Max-Planck-Ring Wiesbaden Germany REF Catalog Number Abbott LOT Batch Code Temperature Limitation Use by ImmunoPlt is a trademark, and CELL-DYN, CELL-DYN Sapphire, and CELL-DYN HemCal are registered trademarks of Abbott Laboratories in various jurisdictions. Manufacturer CELL-DYN ImmunoPlt (CD61) Reagent Page 5 of 6

6 15. DATE OF ISSUANCE E February 2010 CELL-DYN ImmunoPlt (CD61) Reagent Page 6 of 6