December 2010

GP34-A
Validation and Verification of Tubes for

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Venous and Capillary Blood Specimen
Collection; Approved Guideline
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SA

This document provides guidance for conducting validation and

verification testing for venous and capillary blood collection tubes.

A guideline for global application developed through the Clinical and Laboratory Standards Institute consensus process.

Clinical and Laboratory Standards Institute
Setting the standard for quality in clinical laboratory testing around the world.

The Clinical and Laboratory Standards Institute (CLSI) is a not-for-profit membership organization that brings
together the varied perspectives and expertise of the worldwide laboratory community for the advancement of
a common cause: to foster excellence in laboratory medicine by developing and implementing clinical laboratory
standards and guidelines that help laboratories fulfill their responsibilities with efficiency, effectiveness, and
global applicability.

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Consensus—the substantial agreement by materially affected, competent, and interested parties—is core to the
development of all CLSI documents. It does not always connote unanimous agreement, but does mean that the
participants in the development of a consensus document have considered and resolved all relevant objections

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and accept the resulting agreement.

Commenting on Documents

CLSI documents undergo periodic evaluation and modification to keep pace with advancements in technologies,

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procedures, methods, and protocols affecting the laboratory or health care.

CLSI’s consensus process depends on experts who volunteer to serve as contributing authors and/or as
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developed the document is obligated to review all comments, respond in writing to all substantive comments,
and revise the draft document as appropriate.

Comments on published CLSI documents are equally essential, and may be submitted by anyone, at any time, on
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any document. All comments are addressed according to the consensus process by a committee of experts.

Appeals Process

If it is believed that an objection has not been adequately addressed, the process for appeals is documented in
the CLSI Standards Development Policies and Process document.
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 GP34-A
Vol. 30 No. 25
ISBN 1-56238-739-1 Replaces GP34-P
ISSN 0273-3099 Vol. 29 No. 22
Validation and Verification of Tubes for Venous and Capillary Blood
Specimen Collection; Approved Guideline
Volume 30 Number 25
Nancy Dubrowny, MS, MT(ASCP)SC
Elizabeth Armstrong, MT(ASCP)

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Julie Berube, PhD
Raffick A. R. Bowen, MLT(CSMLS), PhD, DCl Chem, FCACB, DABCC
Yung W. Chan, MT(ASCP)
Daniel Hesselgesser, MT(ASCP)
Susan S. Smith
Ana K. Stankovic, MD, PhD, MSPH
Diane I. Szamosi, MA, MT(ASCP)SH
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Abstract
Clinical and Laboratory Standards Institute document GP34-A—Validation and Verification of Tubes for Venous and Capillary
Blood Specimen Collection; Approved Guideline is a guideline for manufacturers of venous and capillary blood collection tubes
and users of blood collection tubes for serum, plasma, and whole blood testing. GP34 provides guidelines for validation and
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verification of test (examination) performance.

Clinical and Laboratory Standards Institute (CLSI). Validation and Verification of Tubes for Venous and Capillary Blood
Specimen Collection; Approved Guideline. CLSI document GP34-A (ISBN 1-56238-739-1). Clinical and Laboratory Standards
Institute, 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087 USA, 2010.

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 revised 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 documents. Current editions are listed in
the CLSI catalog and posted on our website at www.clsi.org. 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:
[email protected]; Website: www.clsi.org.
Number 25 GP34-A

Copyright ©2010 Clinical and Laboratory Standards Institute. Except as stated below, any reproduction of
content from a CLSI copyrighted standard, guideline, companion product, or other material requires
express written consent from CLSI. All rights reserved. Interested parties may send permission requests to
[email protected].

CLSI hereby grants permission to each individual member or purchaser to make a single reproduction of
this publication for use in its laboratory procedure manual at a single site. To request permission to use
this publication in any other manner, e-mail [email protected].

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Suggested Citation

CLSI. Validation and Verification of Tubes for Venous and Capillary Blood Specimen Collection;

Proposed Guideline
October 2009

Approved Guideline
December 2010
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Approved Guideline. CLSI document GP34-A. Wayne, PA: Clinical and Laboratory Standards Institute;
2010.
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ISBN 1-56238-739-1
ISSN 0273-3099

ii
Volume 30 GP34-A

Contents

Abstract ....................................................................................................................................................i

Committee Membership........................................................................................................................ iii

Foreword .............................................................................................................................................. vii

1 Scope .......................................................................................................................................... 1

2 Standard Precautions.................................................................................................................. 1

3 Terminology............................................................................................................................... 1
3.1 A Note on Terminology ................................................................................................ 1

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3.2 Definitions .................................................................................................................... 2
3.3 Abbreviations and Acronyms ....................................................................................... 4
4 Impact of Blood Collection Tubes on Test (Examination) Performance ................................... 4
4.1 Tube Wall .....................................................................................................................5
4.2
4.3
4.4
4.5
4.6
4.7
4.8
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Closures ........................................................................................................................ 5
Closure Lubricant ......................................................................................................... 6
Surfactants .................................................................................................................... 6
Clot Activators .............................................................................................................. 6
Anticoagulants .............................................................................................................. 7
Separator Gel ................................................................................................................8
Trace Metals ................................................................................................................. 8
5 Validation and Verification of Venous Blood Collection Tubes ............................................... 9
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5.1 Preanalytical (Preexamination) Considerations ............................................................ 9
5.2 Determining the Need for Validation and Verification ................................................ 9
5.3 Clinical Evaluation—Planning, Designing, and Conducting the Clinical
Evaluation ................................................................................................................... 10
5.4 Data Analysis .............................................................................................................. 14
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5.5 Clinical Acceptance Criteria ....................................................................................... 15

6 Conclusion ............................................................................................................................... 16

References ............................................................................................................................................. 17

Additional References ........................................................................................................................... 21

Appendix A. Sample Protocol for User Evaluation of Evacuated Venous Blood Collection Tubes .... 23

Appendix B. Example of a Method for Analysis of Precision .............................................................. 31

Summary of Delegate Comments and Subcommittee Responses......................................................... 35

The Quality Management System Approach ........................................................................................ 42

Related CLSI Reference Materials ....................................................................................................... 44

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Volume 30 GP34-A

Foreword
Currently, no guideline is available for either in vitro diagnostic (IVD) manufacturers or clinical
laboratories to validate or verify use of the various venous and capillary blood collection tubes within
each of the following laboratory medicine disciplines: chemistry, immunochemistry, hematology, and
coagulation. However, for microbiology assays or culture methods, several documents address validation
and quality control of collection tubes (see CLSI documents M40, M47, and M15).1-3

This guideline contains information on tubes for venous and capillary blood collection. It is written for
manufacturers of venous and capillary blood collection devices; for assay/instrument manufacturers; and
for those who are responsible for acquisition, handling, and use of the equipment described in this
document.

Specimen collection devices, especially venous and capillary blood collection tubes, are classified as IVD
devices. Because these devices are used to collect patient blood samples that are analyzed on highly

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sensitive clinical instrumentation, it is extremely critical for accurate and precise test results that these
collection devices be verified for use on this instrumentation.

IVD manufacturers are challenged by regulatory agencies to ensure safety and efficacy of their devices as
part of the validation process before release of the devices for use in the clinical laboratory. Tube

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manufacturers can use this guidance document to establish and standardize their validation process for
both current and new blood collection tubes. In addition to this document, CLSI standard H01, Tubes and
Additives for Venous Blood Specimen Collection4—a complementary document to this guideline—details
the requirements for materials, manufacturing, and labeling of blood collection devices.

Additionally, accrediting organizations challenge clinical laboratories to ensure the acceptability or

compatibility of their venous and capillary blood collection devices, with their current instrumentation
and patient population.5 This type of verification will help the clinical laboratories ensure accurate and
precise test results for their collection device and test system.
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Key Words

Capillary blood collection, instrumentation, validation, venous blood collection tubes, verification
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Volume 30 GP34-A

Validation and Verification of Tubes for Venous and Capillary Blood

Specimen Collection; Approved Guideline

1 Scope
This document provides step-by-step recommendations for validation and verification of venous and
capillary blood collection devices. Capillary blood collection devices addressed in this document include
only microcollection devices (see Section 3.2). It also includes guidance for ascertaining the
acceptability/compatibility for clinical performance in chemistry, immunochemistry, hematology, and
coagulation. This guideline does not address validation and verification for clinical performance in
immunohematology, molecular diagnostics, arterial blood gas analysis, proteomics, or genomics.

The focus and procedures of this document are for quantitative measurement only. For qualitative

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measurement, the study requires a different study design.

This document is written for manufacturers of venous and capillary blood collection devices;
assay/instrument manufacturers; all clinical laboratory personnel; and those who are responsible for
acquisition, handling, and use of the equipment described in this document.

2 Standard Precautions
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Because 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 “standard 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 known infectious agents and thus
are more comprehensive than universal precautions, which are intended to apply only to transmission of
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blood-borne pathogens. Standard and universal precaution guidelines are available from the US Centers
for Disease Control and Prevention.6 For specific precautions for preventing the laboratory transmission
of all known infectious agents from laboratory instruments and materials and for recommendations for the
management of exposure to all known infectious disease, refer to CLSI document M29.7

3 Terminology
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3.1 A Note on Terminology

CLSI, as a global leader in standardization, is firmly committed to achieving global harmonization

wherever possible. Harmonization is a process of recognizing, understanding, and explaining differences
while taking steps to achieve worldwide uniformity. CLSI recognizes that medical conventions in the
global metrological community have evolved differently in the United States, Europe, and elsewhere; that
these differences are reflected in CLSI, International Organization for Standardization (ISO), and Comité
Européen de Normalisation (European Committee for Standardization; CEN) documents; and that legally
required use of terms, regional usage, and different consensus timelines are all important considerations in
the harmonization process. In light of this, CLSI’s consensus process for development and revision of
standards focuses on harmonization of terms to facilitate the global application of standards and
guidelines.

In order to align the use of terminology in this document with that of ISO, the terms preexamination,
examination, and postexamination were adopted. For the sake of introduction and to avoid confusion, the
subcommittee chose to include the ISO terms parenthetically where the US terms appear. In addition, the
term sample replaces the term specimen where appropriate, and measurand replaces analyte. The users of

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Clinical and Laboratory Standards Institute. All rights reserved. 1
Number 25 GP34-A

GP34-A should understand that the fundamental meanings of the terms are identical in many cases, and
are defined in the guideline’s Definitions section (see Section 3.2). The terms in this document are
consistent with those defined in the ISO 15189, ISO 17025, and ISO 9000 series of standards.

3.2 Definitions

accuracy (measurement) – closeness of agreement between a measured quantity value and a true
quantity value of a measurand (ISO/IEC Guide 99).8

additive – in a specimen collection tube, any ingredient that is placed in a collection container to facilitate
an intended function (eg, to prevent the blood from clotting or to prevent glycolysis); NOTE: Although
the container closure is not considered an additive, it may contain or be coated with additives, which, if
they come into contact with the specimen, may be considered additives.

analyte – component represented in the name of a measurable quantity (ISO 17511)9; NOTE 1: In the

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type of quantity “mass of protein in 24-hour urine,” “protein” is the analyte. In “amount of substance of
glucose in plasma,” “glucose” is the analyte. In both cases, the long phrase represents the measurand
(ISO 17511)9; NOTE 2: In the type of quantity “catalytic concentration of lactate dehydrogenase
isoenzyme 1 in plasma,” “lactate dehydrogenase isoenzyme 1” is the analyte (ISO 18153).10

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anticoagulant – an agent that prevents coagulation of blood or blood products.

bias – the difference between the expectation of the test results and an accepted reference value (ISO
3534-1)11; NOTE: If the comparison method is a reference method, then the difference between the two
methods measures the trueness of the new method, measured as bias. If the comparison method is not a
reference method, then the trueness of the new method cannot be determined. In this case, one refers to
the difference simply as a difference, and not bias.
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capillary blood – blood obtained by skin puncture or incision that contains a mixture of undetermined
proportions of blood from arterioles, venules, and interstitial and intracellular fluids.

clot activator – material used to initiate the clotting mechanism.

comparative tube – blood collection tube currently used by the clinical laboratory.
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control tube – any reference tube used as a comparative tube when evaluating a new or substantially
modified tube; NOTE: In the United States, these tubes must be US Food and Drug Administration
(FDA) cleared.

draw – quantity of blood drawn into the venous blood collection tube from a venipuncture; NOTE: For
testing purposes, the conditions are defined as follows: 101 kPa (760 mm Hg) pressure and 20 °C ambient
temperature. The temperature of the blood collected is assumed to be 37 °C.

error (measurement) – measured quantity value minus a reference quantity value (ISO/IEC Guide 99).8

expiration date – date after which the product, when stored under recommended conditions, should no
longer be used.

glycolytic inhibitor//antiglycolytic agent – agent that inhibits the use of glucose by blood cells.

imprecision – dispersion of independent results of measurements obtained under specified conditions;

NOTE: It is expressed numerically as standard deviation (SD) or coefficient of variation (CV).

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2 Clinical and Laboratory Standards Institute. All rights reserved.
 Number 25 GP34-A

The Quality Management System Approach

Clinical and Laboratory Standards Institute (CLSI) subscribes to a quality management system approach in the
development of standards and guidelines, which facilitates project management; defines a document structure via a
template; and provides a process to identify needed documents. The approach is based on the model presented in the
most current edition of CLSI document HS01—A Quality Management System Model for Health Care. The quality
management system approach applies a core set of “quality system essentials” (QSEs), basic to any organization, to
all operations in any health care service’s path of workflow (ie, operational aspects that define how a particular
product or service is provided). The QSEs provide the framework for delivery of any type of product or service,
serving as a manager’s guide. The QSEs are:

Documents and Records Equipment Information Management Process Improvement

Organization Purchasing and Inventory Occurrence Management Customer Service
Personnel Process Control Assessments—External Facilities and Safety
and Internal

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GP34-A addresses the QSEs indicated by an “X.” For a description of the other documents listed in the grid, please
refer to the Related CLSI Reference Materials section on the following page.
and Inventory

Improvement

Facilities and
Organization

Management

Management
and Records

and Internal
Information

Assessment
Occurrence
Documents

Purchasing
Equipment

—External
Personnel

Customer
Control
Process

Process

Service

Safety
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X
EP05
EP09
EP10
EP15
EP21
H01
H03
EP10

H03
H04
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H18
H21
M15 M29

M40
M47
Adapted from CLSI document HS01—A Quality Management System Model for Health Care.
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42 Clinical and Laboratory Standards Institute. All rights reserved.
Volume 30 GP34-A

Path of Workflow

A path of workflow is the description of the necessary steps to deliver the particular product or service that the
organization or entity provides. For example, CLSI document GP26⎯Application of a Quality Management System
Model for Laboratory Services defines a clinical laboratory path of workflow, which consists of three sequential
processes: preexamination, examination, and postexamination. All clinical laboratories follow these processes to
deliver the laboratory’s services, namely quality laboratory information.

GP34-A addresses the clinical laboratory path of workflow steps indicated by an “X.” For a description of the other
documents listed in the grid, please refer to the Related CLSI Reference Materials section on the following page.

Preexamination Examination Postexamination

Examination ordering

Results reporting and

Sample management
receipt/processing
Sample collection

Sample transport

Results review
and follow-up

Interpretation
Examination

archiving
Sample

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X
H01
H03 H03
H04

H21
M15

M47
H03

H18
H21
M15
M40
M47
PLH03

H18

M15

M47
Adapted from CLSI document HS01—A Quality Management System Model for Health Care.
H03

H18

M15

M47
H03

M15

M47
M15

M47 M47
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Clinical and Laboratory Standards Institute. All rights reserved. 43
Number 25 GP34-A

Related CLSI Reference Materials∗

EP05-A2 Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline—

Second Edition (2004). This document provides guidance for designing an experiment to evaluate the
precision performance of quantitative measurement methods; recommendations on comparing the resulting
precision estimates with manufacturers’ precision performance claims and determining when such
comparisons are valid; as well as manufacturers’ guidelines for establishing claims.

EP09-A2-IR Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline—Second Edition
(Interim Revision) (2010). This document addresses procedures for determining the bias between two clinical
methods, and the design of a method comparison experiment using split patient samples and data analysis.

EP10-A3 Preliminary Evaluation of Quantitative Clinical Laboratory Measurement Procedures; Approved

Guideline—Third Edition (2006). This guideline provides experimental design and data analysis for
preliminary evaluation of the performance of a measurement procedure or device.

EP15-A2 User Verification of Performance for Precision and Trueness; Approved Guideline—Second Edition

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(2005). This document describes the demonstration of method precision and trueness for clinical laboratory
quantitative methods using a protocol designed to be completed within five working days or less.

EP21-A Estimation of Total Analytical Error for Clinical Laboratory Methods; Approved Guideline (2003).
This document provides manufacturers and end users with a means to estimate total analytical error for an
assay. A data collection protocol and an analysis method that can be used to judge the clinical acceptability of

H01-A5

H03-A6
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new methods using patient specimens are included. These tools can also monitor an assay’s total analytical
error by using quality control samples.

Tubes and Additives for Venous Blood Specimen Collection; Approved Standard—Fifth Edition (2003).
This document contains requirements for venous blood collection tubes and additives, including technical
descriptions of ethylenediaminetetraacetic acid (EDTA), sodium citrate, and heparin compounds used in blood
collection devices.

Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture; Approved Standard—
Sixth Edition (2007). This document provides procedures for the collection of diagnostic specimens by
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venipuncture, including line draws, blood culture collection, and venipuncture in children.

H04-A6 Procedures and Devices for the Collection of Diagnostic Capillary Blood Specimens; Approved
Standard—Sixth Edition (2008). This document provides a technique for the collection of diagnostic
capillary blood specimens, including recommendations for collection sites and specimen handling and
identification. Specifications for disposable devices used to collect, process, and transfer diagnostic capillary
blood specimens are also included.
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H18-A4 Procedures for the Handling and Processing of Blood Specimens for Common Laboratory Tests;
Approved Guideline—Fourth Edition (2010). This document includes criteria for preparing an optimal
serum or plasma sample and for the devices used to process blood specimens.

H21-A5 Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation
Assays and Molecular Hemostasis Assays; Approved Guideline—Fifth Edition (2008). This document
provides procedures for collecting, transporting, and storing blood; processing blood specimens; storing
plasma for coagulation testing; and general recommendations for performing the tests.

M15-A Laboratory Diagnosis of Blood-borne Parasitic Diseases; Approved Guideline (2000). This document
provides guidance on specimen collection, optimum timing for preparing blood films, blood film preparations,
staining procedures, examination of specimens, and identification of parasites.

∗
CLSI documents are continually reviewed and revised through the CLSI consensus process; therefore, readers should refer to
the most current editions.

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44 Clinical and Laboratory Standards Institute. All rights reserved.
Volume 30 GP34-A

Related CLSI Reference Materials (Continued)

M29-A3 Protection of Laboratory Workers From Occupationally Acquired Infections; Approved Guideline—
Third Edition (2005). Based on US regulations, this document provides guidance on the risk of transmission
of infectious agents by aerosols, droplets, blood, and body substances in a laboratory setting; specific
precautions for preventing the laboratory transmission of microbial infection from laboratory instruments and
materials; and recommendations for the management of exposure to infectious agents.

M40-A Quality Control of Microbiological Transport Systems; Approved Standard (2003). This document
provides criteria to assist manufacturers and end users of transport devices in providing and selecting
dependable products for the transport of microbiological clinical specimens.

M47-A Principles and Procedures for Blood Cultures; Approved Guideline (2007). This document provides
recommendations for the collection, transport, and processing of blood cultures as well as guidance for the
recovery of pathogens from blood specimens taken from patients who are suspected of having bacteremia or
fungemia.

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Clinical and Laboratory Standards Institute. All rights reserved. 45
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