ISO 16140-2:2016

Overview - ISO 16140-2:2016 (Method validation for microbiology of the food chain)

ISO 16140-2:2016 establishes a harmonized protocol for the validation of alternative (mostly proprietary) microbiological methods against a reference method. It is part of the ISO 16140 series (Microbiology of the food chain - Method validation) and is intended for organizations conducting validation studies. The standard covers both qualitative (detection) and quantitative (enumeration) methods used for microorganisms in foods, feeds, environmental samples in production/processing, and primary production samples. It is especially applicable to bacteria and fungi, with some clauses applicable to other organisms (viruses, parasites) on a case-by-case basis.

Key technical topics and requirements

• Two-phase validation approach
  • Method comparison study (performed in the organizing laboratory)
  • Interlaboratory study (multi-lab evaluation)
• Protocols for qualitative and quantitative methods
  • Qualitative: sensitivity study, Relative Level of Detection (RLOD), inclusivity/exclusivity testing
  • Quantitative: relative trueness, accuracy profile, limit of quantification (LOQ), inclusivity/exclusivity
• Paired vs unpaired study designs
  • Guidance when methods share the same initial test portion (paired) or require separate portions (unpaired)
• Statistical and interpretative tools
  • Use of Acceptability Limits (AL), RLOD models, accuracy profile methodology and interlaboratory variance assessment
• Annexes and supporting guidance
  • Classification of sample types, contamination protocols, strain selection criteria, RLOD calculation models, and worked examples of accuracy profile application

Practical applications - who uses ISO 16140-2:2016

• Method developers validating proprietary rapid tests or novel assays for market adoption
• Independent validation bodies and accredited laboratories conducting method validation studies
• Food and feed industry laboratories assessing alternative methods for routine monitoring and quality control
• Regulators and certification bodies evaluating method performance data for acceptance or recognition
• Researchers and contract testing labs designing comparative studies and interlaboratory trials

Using ISO 16140-2 helps produce reproducible, auditable validation data for adoption of faster or easier alternatives to reference methods, supporting risk-based testing and regulatory acceptance.

Related standards

• ISO 16140-1 - Vocabulary for method validation (normative reference)
• Other parts of ISO 16140 (in preparation): verification, single-lab validation, factorial interlaboratory validation, and validation for confirmation/typing

Keywords: ISO 16140-2:2016, method validation, microbiology of the food chain, alternative methods, proprietary methods, method comparison study, interlaboratory study, RLOD, accuracy profile, inclusivity exclusivity.