totalβhcg(5this).docVIP

Total βhCG (5th is) ASSAY PROCEDURE Standardized to the WHO 5th International Standard for Chorionic Gonadotropin FOR USE WITH TEST NAMEs: HCG5, HCG5d and d-CG5 total βhCG QUANTITATIVE DETERMINATION IN HUMAN SERUM AND PLASMA BY THE ACCESS IMMUNOASSAY SYSTEMS Table of Contents Principle 2 Specimen Collection 2 Reagents and Equipment 3 Calibration Details 5 Quality Control 6 Procedure 6 Results 6 Procedural Comments 7 Limitations of the Procedure 7 References 10 This document is provided as an aid to writing laboratory procedures following CLSI guidelines but does not include all activities identified in CLSI GP2-A5 Clinical Laboratory Technical Procedure Manuals; Approved Guideline – Fifth Edition. Each laboratory is responsible for ensuring their procedures are comprehensive and complete. Beckman Coulter, Inc. ( Brea, CA 92821 Principle Principles of the Procedure The Access Total βhCG (5th IS) assay is a sequential two-step immunoenzymatic (“sandwich”) assay. A sample is added to a reaction vessel along with a citrate buffer. After an initial incubation, rabbit anti-βhCG alkaline phosphatase conjugate and paramagnetic particles coated with goat anti-mouse IgG: mouse monoclonal anti-βhCG complexes are added. The hCG binds to the immobilized monoclonal anti-βhCG on the solid phase while, at the same time, the rabbit anti-βhCG alkaline phosphatase conjugate reacts with different antigenic sites on the hCG. After incubation in a reaction vessel, materials bound to the solid phase are held in a magnetic field while unbound materials are washed away. Then, the chemiluminescent substrate Lumi-Phos* 530 is added to the vessel and light generated by the reaction is measured with a luminometer. The light production is directly proportional to the concentration of total βhCG in the sample. The amount of analyte in the sample is determined from a stored, multi-point calibration curve. Summary and Explanation Human chorionic gonadot