Which CLSI EP protocol do you need? A map of the EP protocols by the question each one answers, and whether your situation calls for establishing performance or verifying it. Start here and follow the links to the detail.

The CLSI EP series covers quantitative and qualitative method validation end to end. The naming (EP05, EP09, EP17, EP28) tells you nothing about which one your task needs. This page maps the protocols to the questions they answer. Start from “I need to know X about my method” and arrive at the right procedure, and the right depth of study, with a guide for each step.

First: are you establishing or verifying?

Before choosing a protocol, settle the question that determines how much work each one is: are you validating a method from scratch or verifying a manufacturer’s claims? An unmodified, cleared assay used as intended needs verification: short studies confirming stated performance. A laboratory-developed or modified method needs validation: fuller studies establishing performance yourself. The same characteristic (precision, say) has a short verification protocol and a full establishment protocol. Picking the wrong one wastes weeks or leaves a gap. This decision runs through everything below.

The protocols by question

The question Protocol Guide
Is this method even worth full validation? EP10-A3 (preliminary) Rapid method evaluation
How precise is it? (establish) EP05-A3 Establishing precision
Does the claimed precision hold here? (verify) EP15-A3 Verifying precision claims
Does it agree with another method? EP09-A3 Choosing a regression · Bland–Altman
What is the total error of a result? EP21-A Total analytical error
Over what range is it linear? EP06-Ed2 Assessing linearity · measuring interval
What is the lowest it can detect or measure? EP17-A2 LoB, LoD and LoQ
Does a substance interfere with the result? EP07-Ed3 Interference and recovery testing
What is the reference interval? EP28-A3C Choosing a method
How well does it discriminate disease? EP24-A2 ROC and AUC
How accurate is a qualitative test? EP12-A2 Evaluating a qualitative test
Does a reference material behave like a patient sample? EP14-A3 Commutability

The protocols by number

The table above works forwards, from a question to a protocol. Most people arrive with the number instead, having seen it in an inspection finding, a supplier’s documentation or a colleague’s protocol. Here is the same set the other way round.

Protocol Title, in short Guide
EP05Precision of quantitative measurement procedures — establishing itEstablishing precision
EP06Linearity of a quantitative measurement procedureAssessing linearity
EP07Interference testing in clinical chemistryInterference and recovery testing
EP09Measurement procedure comparison and bias estimation using patient samplesChoosing a regression
EP10Preliminary evaluation of a quantitative measurement procedureRapid method evaluation
EP12Evaluation of qualitative, binary output examination performanceEvaluating a qualitative test
EP14Commutability of processed samplesCommutability
EP15User verification of precision and estimation of biasVerifying precision claims
EP17Detection capability — limit of blank, detection and quantitationLoB, LoD and LoQ
EP21Total analytical error for quantitative measurement proceduresTotal analytical error
EP24Assessment of the diagnostic accuracy of laboratory tests using ROC curvesROC curves and AUC
EP28Defining, establishing and verifying reference intervalsChoosing a reference interval method
EP31Verification of comparability of patient results within one health care systemComparing instruments within one laboratory

Editions change, so check the current designation on the CLSI catalogue before citing one in a validation file. EP05 reached its fourth edition in December 2025 and EP21 its third in June 2025, while EP15-A3, EP17-A2 and EP28-A3c remain the current versions of theirs.

A typical order of work

For a new quantitative method being validated in full, the studies fall into a natural sequence. A short preliminary evaluation screens out gross problems first. Then precision and linearity characterise the method on its own. Detection capability fixes the low end.

A method comparison then places it against an existing method, yielding the bias at decision points and, if needed, the total error. Finally the reference interval is established or transferred. For a diagnostic claim the diagnostic performance is characterised. Verifying an unmodified cleared assay is the same map with most steps shortened to confirmations.

A vertical numbered pipeline of six validation stages: 1 preliminary screen (EP10-A3), 2 precision and linearity (EP05-A3, EP06-Ed2), 3 detection capability (EP17-A2), 4 method comparison (EP09-A3) with an optional Total error branch (EP21), 5 reference interval (EP28-A3C), and 6 diagnostic performance (EP24-A2, EP12-A2), shown dashed as conditional.
The usual sequence for validating a new quantitative method in full. A preliminary screen comes first. Precision, linearity and detection capability characterise the method on its own. A comparison then places it against an existing method. Then the reference interval, and for a diagnostic claim diagnostic performance. Verifying a cleared assay follows the same map with most steps shortened to confirmations.

The specifications behind every judgement

Every protocol above produces a number that has to be judged against a limit. Those limits deserve as much thought as the measurements. Where the acceptance criteria come from is its own subject: analytical performance specifications derived from biological variation or clinical outcome, and the allowable total error a result is judged against. Getting the specification right is what makes a measured number a valid pass or fail. Settle the distinction between trueness, precision and accuracy early, because every protocol depends on it.

Work through the protocols with Analyse-it

Analyse-it performs the statistical analysis behind most of the protocols on this map, inside Excel:

Every feature from all five editions for 15 days. Across the Medical, Method Validation and Ultimate editions, from US$ 340 a year. Validated against NIST and CLSI reference datasets. If you are starting from a regulatory obligation rather than a statistical question, what CLIA requires before you report a result, CAP accreditation and AMR verification, performance studies for an IVD 510(k) and method validation under ICH Q2(R2) come at the same map from that direction. Every protocol here is one part of a measurement system analysis, and what goes in a validation report covers the document they all end up in.