ISO 16297:2020

Overview

ISO 16297:2020 - Milk - Bacterial count - Protocol for the evaluation of alternative methods specifies a statistical and practical protocol for evaluating instrumental alternative methods used to determine the total bacterial count in raw milk (animals of different species). It is intended to assess whether an alternative method gives reliable, fit-for-purpose results compared with an anchor method (an internationally recognized reference method). The document complements ISO 16140-2 and ISO 8196 | IDF 128 and is aimed at laboratories, manufacturers and regulators involved in milk microbiology and food safety.

Key topics and technical requirements

The standard lays out the attributes and tests required to demonstrate performance of an alternative method, including:

• Method description & checklist
  • Principle, parameter/unit, technical design, field of application (purpose, matrix), suppliers and method specification (sampling prerequisites, preservation, measurement spectrum, precision and accuracy statements, throughput).
• Data transformation
  • Use of transformations (commonly logarithmic) to approximate normal distributions for microbiological counts before statistical analysis.
• Measuring range
  • Lower limit of quantification (LLOQ) determined from repeated low-count milk samples (standard deviation estimated; generally n = 20) and calculated according to ISO 16140-2.
  • Upper limit of quantification defined by maximum reliable instrument reading and methodological limitations (e.g., coincidence effects).
  • Linearity of instrument signal across the measuring range.
• Analytical attributes
  • Carry-over, stability, precision (repeatability and reproducibility), and factors affecting results.
• Estimation vs. anchor method
  • Protocols for measurement, calculations (including visual scatter checks, outlier handling), accuracy profile and how to express attributes in anchor-method units.
• Performance rating
  • Procedures to rate the elaborated attributes and decide fitness for intended use.

Practical applications

ISO 16297:2020 is used to validate and document the performance of instrumental alternative methods such as flow cytometry or other automated counting technologies (as compared with plate counts). Typical applications include:

• Validating new microbiological instruments for routine milk quality control and milk grading
• Demonstrating equivalence to regulatory or contractual anchor methods
• Establishing measurement ranges, LLOQ/ULOQ and instrument linearity for method implementation
• Supporting method adoption by dairies, testing labs and instrument suppliers

Who should use this standard

• Dairy testing laboratories and quality control teams
• Instrument and reagent manufacturers developing alternative bacterial count methods
• Regulatory authorities and accreditation bodies assessing method equivalence
• Researchers and consultants involved in method validation for raw milk microbiology

Related standards

• ISO 16140-2 (method validation - comparison protocol)
• ISO 8196 | IDF 128 series (evaluation of overall accuracy)
• ISO 5725 parts (trueness and precision)
• ISO 21187 | IDF 196 (conversion relationships between alternative and anchor methods)

Keywords: ISO 16297:2020, milk bacterial count, total bacterial count, alternative methods, raw milk, method validation, anchor method, LLOQ, instrument linearity, precision, accuracy profile.