ISO 16140-3-2021 PDF

Full title and description

Microbiology of the food chain — Method validation — Part 3: Protocol for the verification of reference methods and validated alternative methods in a single laboratory. This international standard specifies the protocol to be used by a single laboratory to verify that a reference method or a validated alternative method performs as expected when implemented in the user laboratory environment.

Abstract

This document provides technical protocols and requirements for the verification (single-laboratory) of microbiological methods used for analysis (detection and/or quantification), confirmation and typing of microorganisms in products for human consumption, products for animal feeding, environmental samples related to food and feed production/handling and samples from primary production. It is primarily applicable to bacteria and fungi; some clauses may be applied to other microorganisms or their metabolites on a case-by-case basis. The standard describes specific technical protocols for validated qualitative methods, validated quantitative methods and validated alternative confirmation and typing methods, and includes provisions (Annex F) for verification of non-validated reference methods during a defined transition period.

General information

• Status: Published.
• Publication date: January 2021 (published 19 January 2021).
• Publisher: International Organization for Standardization (ISO).
• ICS / categories: 07.100.30 — Microbiology of food chain / Method validation.
• Edition / version: Edition 1 (2021).
• Number of pages: 70 pages (main publication).

Scope

ISO 16140-3:2021 applies to the single‑laboratory verification of both reference methods and previously validated alternative methods used for detection, enumeration, confirmation and typing of microorganisms in food, feed, environmental and primary‑production samples. The document provides distinct technical protocols for validated qualitative and quantitative methods and for validated alternative confirmation/typing methods, and it includes a procedure for verifying non‑validated reference methods during a transition period (see Annex F). The standard is intended to support laboratory implementation and accreditation activities (e.g., ISO/IEC 17025) by defining performance checks, acceptance criteria and documentation requirements.

Key topics and requirements

• Protocol for verification of validated qualitative methods (technical requirements and acceptance criteria).
• Protocol for verification of validated quantitative methods (statistical approaches, performance characteristics to assess: precision, trueness, limits).
• Protocol for verification of validated alternative confirmation and typing methods.
• Procedure for verification of non‑validated reference methods (Annex F) applicable during the transition period.
• Requirements for documentation, test sample selection, comparator/reference method use, statistical evaluation and record keeping to demonstrate equivalent performance in the user laboratory.
• Guidance intended to support accreditation and consistent implementation across laboratories (ties to ISO/IEC 17025 principles).

Typical use and users

Primary users are food and feed testing laboratories, public health and regulatory laboratories, quality control laboratories within industry, method developers and manufacturers of alternative microbiological methods. Accreditation bodies, proficiency testing providers and laboratories preparing for or maintaining ISO/IEC 17025 accreditation commonly use ISO 16140-3 as the reference for single‑laboratory verification procedures.

Related standards

ISO 16140-3:2021 is part of the ISO 16140 series on microbiological method validation. Relevant related publications include ISO 16140-1 (vocabulary), ISO 16140-2 (validation of alternative/proprietary methods), ISO 16140-4 (protocol for method validation in a single laboratory) and other parts of the series that define validation and verification terminology and study designs. Laboratories implementing verification should also consider ISO/IEC 17025 (general requirements for the competence of testing and calibration laboratories) for accreditation context.

Keywords

method validation, method verification, microbiology, food chain, single‑laboratory verification, reference method, alternative method, qualitative method, quantitative method, confirmation, typing, performance characteristics, ISO 16140.

FAQ

Q: What is this standard?

A: ISO 16140-3:2021 is an international standard that specifies the protocol for verification of reference methods and validated alternative microbiological methods when implemented in a single (user) laboratory.

Q: What does it cover?

A: It covers technical procedures and acceptance criteria for verifying validated qualitative and quantitative microbiological methods (detection, enumeration), validated alternative confirmation and typing methods, and includes a procedure for verification of non‑validated reference methods (Annex F) applicable during the transition period.

Q: Who typically uses it?

A: Food/feed testing laboratories, regulatory/public health laboratories, quality control laboratories in industry, method developers and accreditation bodies use the standard to verify method performance prior to routine use and/or for accreditation purposes.

Q: Is it current or superseded?

A: ISO 16140-3:2021 is the current published edition (first edition: January 2021). The standard is subject to periodic review and has an amendment published in 2025 addressing verification of validated identification methods; non‑validated reference method verification provisions are transitional (see TC 34/SC 9 transition guidance).

Q: Is it part of a series?

A: Yes — it is part of the ISO 16140 series on method validation (e.g., ISO 16140-1, -2, -4 among others) that together define vocabulary, validation and verification protocols for microbiological methods in the food chain.

Q: What are the key keywords?

A: Method verification, validation, microbiology, food, feed, single‑laboratory, reference method, alternative method, qualitative, quantitative, performance characteristics.