Standards Development

Scope

(Please see ISO/IEC Directives, Part 1, Annex C, Clause C.4.3)

This document specifies considerations and requirements for handling, measuring, and storing ultralow concentration of target nucleic acid sequences, i.e., concentrations corresponding to copy numbers that are significantly affected by Poisson distribution.

This document is applicable to qPCR, dPCR, isothermal amplification, microarray, and NGS.

This document can be used in molecular biology labs. It can also be used in accredited laboratories such as ISO 17025.

Purpose

(Please see ISO/IEC Directives, Part 1, Annex C and additional guidance on justification statements in the brochure Guidance on New Work)

Analytical methods based on polymerase chain reaction (PCR) are theoretically capable of detecting the presence of a one molecule of nucleic acid in the reaction solution, making it possible to amplify, measure, and quantify the ultra-low concentration target nucleic acid sequence. It is used not only in the field of biotechnology, but also in many other fields such as medicine, food, and the environment. However, it is difficult to stably store, measure, quantify, and detect ultra-low concentration target nucleic acid sequences. For example, nucleic acids readily adsorb to polypropylene (PP), which is widely used as a base material for labware, so target nucleic acid sequences stored in such containers are easily removed from the sample/reaction solution during storage or experiment and become undetectable. Similarly, for example, if a sample before PCR and a sample after PCR are handled in the same space for the same target nucleic acid sequence, contamination will occur, and the measurement result will change. Furthermore, when the target nucleic acid sequence is present at ultra-low concentrations, the distribution of nucleic acid molecules becomes non-uniform. These phenomena may occur even in samples of general concentrations, but at such concentrations, the effects of adsorption and contamination are relatively small and negligible. However, since ultra-low concentration samples have a significant impact on the measurement results, there are special considerations to be taken into account when dealing with ultra-low concentration samples.

This document provides specific requirements to improve data reliability and obtain accurate measurement results when working with ultra-low concentrations of target nucleic acid sequence. Biotechnology and bioscience industry data with higher measurement confidence will enable data interoperability, improved product quality, reduced risks and costs and facilitate international trade.