1、H21-A5Vol. 28 No. 5Replaces H21-A4Vol. 23 No. 35Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays; Approved Guideline Fifth EditionThis document provides procedures for collecting, transporting, and storing blood; pro

2、cessing blood specimens; storing plasma for coagulation testing; and general recommendations for performing the tests.A guideline for global application developed through the Clinical and Laboratory Standards Institute consensus process.CLINICAL ANDLABORATORYSTANDARDSINSTITUTE(Formerly NCCLS)Volume

3、28 Number 5H21-A5ISBN 1-56238-657-3ISSN 0273-3099Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays; Approved GuidelineFifth EditionDorothy M. Adcock, MDDaniel M. Hoefner, MT, PhDKandice Kottke-Marchant, MD, PhDRichard

4、 A. Marlar, PhDDiane I. Szamosi, MA, MT(ASCP), SH(ASCP)David J. Warunek, PhD, MBAAbstractClinical and Laboratory Standards Institute H21-A5Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays; Approved GuidelineFifth Edi

5、tion is an update of the previous edition published in 2003. The guideline provides procedures for the collection, transport, and processing of blood specimens for plasma-based and molecular coagulation testing. Tests of the coagulation system are very sensitive to storage (time and temperature), co

6、ncentration of anticoagulant, and surface of containers; attention to these parameters is important. H21-A5 is primarily directed toward laboratory and/or clinical personnel responsible for obtaining patient specimens and preparing samples for plasma-based or molecular coagulation testing.Clinical a

7、nd Laboratory Standards Institute (CLSI). Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays; Approved GuidelineFifth Edition. CLSI document H21-A5 (ISBN 1-56238-657-3). Clinical and Laboratory Standards Institute, 940

8、 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 2008.The Clinical and Laboratory Standards Institute consensus process, which is the mechanism for moving a document through two or more levels of review by the health care community, is an ongoing process. Users should expect revise

9、d editions of any given document. Because rapid changes in technology may affect the procedures, methods, and protocols in a standard or guideline, users should replace outdated editions with the current editions of CLSI/NCCLS documents. Current editions are listed in the CLSI catalog and posted on

10、our website at . If your organization is not a member and would like to become one, and to request a copy of the catalog, contact us at: Telephone: 610.688.0100; Fax: 610.688.0700; E-Mail: ; Website: (Formerly NCCLS)Volume 28H21-A5ContentsAbstract .iCom

11、mittee Membership.iiiForeword .vii1Scope.12Introduction.13Standard Precautions.14Definitions .15Specimen Collection .35.1Specimen Labeling .35.2Methods for Obtaining Specimens.45.3Specimen Collection Containers and Additives.75.4Needle Gauge.96Specimen Transport, Processing, and Sample Storage .106.

12、1Specimen Transport .106.2Processing Suitable Specimens for Plasma-Based Coagulation Assays.116.3Processing Suitable Specimens for Molecular Hemostasis Assays .127Storage .137.1For Plasma-Based Coagulation Assays .137.2For Molecular Hemostasis Assays.168Summary of Causes for Specimen Rejection.168.1

13、For Plasma-Based Coagulation Assays .168.2For Molecular Hemostasis Assays.17References.18Appendix A. Amount of Anticoagulant Solution/Volume of Blood at Different PackedCell Volume Values.22Appendix B. Summary of Data Showing the Variation of Results From the Baseline ValueWith Time of All Analytes

14、Measured.23Summary of Consensus Comments and Committee Responses .24Summary of Delegate Comments and Subcommittee Responses.26The Quality Management System Approach .32Related CLSI Reference Materials .33vVolume 28H21-A5ForewordBecause of the many variables that can affect coagulation test results,

15、CLSI has made available this guideline, which describes procedures for collection, transport, preparation, and storage of samples for plasma-based coagulation assays and molecular hemostasis testing. This publication should enhance the uniformity of sample collection, preparation, and handling and,

16、thereby, reduce many of the preanalytical variables that can affect the test results.This document replaces the fourth edition of the approved guideline, H21-A4, which was published in 2003. Several changes were made in this edition; chief among them is the revision of transportation and storage gui

17、delines for plasma -based hemostasis testing and the addition of information pertinent to the collection, transportation, and processing of specimens for molecular hemostasis assays.Key WordsActivated partial thromboplastin time, citrate, coagulation, preanalytical variables, prothrombin time, sampl

18、e storage, specimen collection, specimen transportviiVolume 28H21-A5Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays; Approved GuidelineFifth Edition1ScopeThis guideline covers the procedures for the collection, tran

19、sport, and processing of specimens for plasma- based coagulation and molecular hemostasis tests. Many variables, including anticoagulant volume and concentration, type of tube additive, duration and temperature of specimen storage, and surface of containers used for specimen collection and storage,

20、may affect plasma-based coagulation test results. The reliability and accuracy of molecular test results also depend upon a variety of specimen collection, transport, and storage factors. The molecular testing in this document refers to DNA testing only.The document is directed toward laboratory and

21、/or clinical personnel responsible for obtaining and preparing patient specimens and for plasma-based coagulation and molecular hemostasis testing. It is also aimed at manufacturers of products involved in specimen collection, storage, preparation, and testing of plasma-based or molecular hemostasis

22、 assays. This document does not address whole blood clotting tests, platelet function tests, or point-of-care testing. H21-A5 does not provide general guidelines for the performance of coagulation testing. Performance guidelines for specific coagulation assays are addressedin other CLSI documents, s

23、uch as those for PT and APTT assays (ie, H471) and fibrinogen assay (ie, H302).2IntroductionA procedural guideline for the collection, transport, and processing of specimens for plasma-based coagulation and molecular hemostasis tests is necessary, as many preanalytical variables may affect test resu

24、lts (eg, concentration and volume of anticoagulant or additive; specimen and sample storage time and temperature). Because important diagnostic and therapeutic decisions are based on the results of hemostasis assays, a procedural guideline for the collection, transport, and processing of specimens f

25、or the general performance of plasma-based coagulation and molecular hemostasis assays is warranted.3Standard PrecautionsBecause it is often impossible to know what isolates or specimens might be infectious, all patient and laboratory specimens are treated as infectious and handled according to “sta

26、ndard precautions.” Standard precautions are guidelines that combine the major features of “universal precautions and body substance isolation” practices. Standard precautions cover the transmission of all infectious agents and thus are more comprehensive than universal precautions, which are intend

27、ed to apply only to transmission of blood-borne pathogens. Standard and universal precaution guidelines are available from the US Centers for Disease Control and Prevention.3 For specific precautions for preventing the laboratory transmission of all infectious agents from laboratory instruments and

28、materials and for recommendations for the management of exposure to all infectious disease, refer to CLSI document M29.44Definitionsactivated partial thromboplastin time (APTT) the time, in seconds, required for a fibrin clot to form in a plasma sample after appropriate amounts of calcium chloride,

29、and a partial thromboplastin reagent (phospholipid plus a contact activator), are mixed with the sample; NOTE: The APTT measures the intrinsic and common coagulation pathways.Clinical and Laboratory Standards Institute. All rights reserved.1Volume 28H21-A5Related CLSI Reference MaterialsC28-A2How to

30、 Define and Determine Reference Intervals in the Clinical Laboratory; Approved GuidelineSecond Edition (2000). This document provides guidance for determining reference values and referenceintervals for quantitative clinical laboratory tests.GP2-A5Laboratory Documents: Development and Control; Appro

31、ved GuidelineFifth Edition (2006). Thisdocument provides guidance on development, review, approval, management, and use of policy, process, andprocedure documents in the medical laboratory community.H1-A5Tubes and Additives for Venous Blood Specimen Collection; Approved StandardFifth Edition (2003).

32、This document contains requirements for venous blood collection tubes and additives, including technicaldescriptions of ethylenediaminetetraacetic acid (EDTA), sodium citrate, and heparin compounds used in bloodcollection devices.H3-A6Procedures for the Collection of Diagnostic Blood Specimens by Ve

33、nipuncture; Approved StandardSixth Edition (2007). This document provides procedures for the collection of diagnostic specimens byvenipuncture, including line draws, blood culture collection, and venipuncture in children.H30-A2Procedure for the Determination of Fibrinogen in Plasma; Approved Guideli

34、neSecond Edition(2001). This document provides general guidelines for performing the fibrinogen assay in the clinicallaboratory. It also includes reporting of results and in vivo and in vitro conditions that may alter results.H47-AOne-Stage Prothrombin Time (PT) Test and Activated Partial Thrombopla

35、stin Time (APTT) Test;Approved Guideline (1996). This document provides guidelines for performing the PT and APTT tests in theclinical laboratory, for reporting results, and for identifying sources of error.H51-AAssays of von Willebrand Factor Antigen and Ristocetin Cofactor Activity; Approved Guide

36、line (2002).This guideline describes the following: appropriate test specimens; reagents and materials; methods of plateletagglutination and ELISA; preparation of reference curves; determination of reference intervals; quality controlprocedures; result interpretation; and sources of error for assays

37、 of von Willebrand factor antigen andristocetin cofactor activity. A brief description of von Willebrand disease and its various subtypes is included,as well as a list of references to more comprehensive reviews of this commonly inherited and rarely acquiredbleeding disorder.M29-A3Protection of Labo

38、ratory Workers From Occupationally Acquired Infections; Approved GuidelineThird Edition (2005). Based on US regulations, this document provides guidance on the risk of transmissionof infectious agents by aerosols, droplets, blood, and body substances in a laboratory setting; specificprecautions for preventing the laboratory transmission of microbial infection from laboratory instruments andmaterials; and recommendations for the management of exposure to infectious agents.MM1-A2Molecular Diagnostic Methods for Genetic Diseases; Approved Guidelin