What is ISO 20395 - Performance of quantification method for qPCR and dPCR about?
ISO 20395 discusses biotechnology. ISO 20395 is an international standard on performance evaluation of quantification methods for nucleic acid target sequences — qPCR and dPCR in biotechnology.
ISO 20395 details generic requirements for evaluating the performance and ensuring the quality of methods used for the quantification of specific nucleic acid sequences (targets).
ISO 20395 specifies:
- Analytical design including quantification strategies and use of controls
- Quantification of total nucleic acid mass concentration and quality control of a nucleic acid sample including assessment of nucleic acid quality (purity and integrity)
- PCR assay design, optimization, in silico and in vitro specificity testing
- Data quality control and analysis including acceptance criteria, threshold setting and normalization
- Method validation with specific requirements for qPCR and dPCR
- Approaches to establishing metrological traceability and estimating measurement uncertainty
ISO 20395 applies to the quantification of DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) target sequences, using either digital (dPCR) or quantitative real-time PCR (qPCR) amplification technologies.
ISO 20395 also applies to target sequences present in nucleic acid molecules including double-stranded DNA (dsDNA) such as genomic DNA (gDNA) and plasmid DNA, single-stranded DNA (ssDNA), complementary DNA (cDNA), and single-stranded RNA (ssRNA) including ribosomal RNA (rRNA), messenger RNA (mRNA), and long and short non-coding RNA [microRNAs (miRNAs) and short interfering RNAs (siRNAs)], as well as double-stranded RNA (dsRNA).
ISO 20395 also applies to nucleic acids derived from biological sources such as viruses, prokaryotic and eukaryotic cells, cell-free biological fluids (e.g., plasma or cell media), or in vitro sources [e.g., oligonucleotides, synthetic gene constructs, and in vitro transcribed (IVT) RNA].
Note 1: ISO 20395 does not apply to the quantification of very short DNA oligonucleotides (<50 bases).
Note 2: ISO 20395 does not provide requirements or acceptance criteria for the sampling of biological materials or processing of biological samples (i.e., collection, preservation, transportation, storage, treatment, and nucleic acid extraction). Nor does it provide requirements and acceptance criteria for specific applications (e.g., food or clinical applications where specific matrix issues can arise).
Who is ISO 20395 - Performance of quantification method for qPCR and dPCR for?
ISO 20395 on quantification methods for nucleic acid target sequences — qPCR and dPCR is useful for:
- Nanotechnology test laboratories
- Nanotechnology developers
- Regulatory authorities
Why should you use ISO 20395 –Performance of quantification method for qPCR and dPCR?
Both quantitative PCR (qPCR) and digital PCR(dPCR) provide sensitive and specific detection, and precise quantification of nucleic acids.
ISO 20395 has been developed to specifically support the analytical requirements withrespect to the quantification of specific nucleic acid sequences (targets). Quantification of nucleic acid target sequences is a cross-cutting fundamental measurement that broadly impacts many aspects of biotechnology.
ISO 20395 guidelines assist you with the design of measurement procedure, total nucleic acid quantity, integrity, purity, optimization for quantification of nucleic acid target sequences, data quality control (qc), nucleic acid quantification measurement method validation, nucleic acid quantification measurement traceability, comparability, and measurement uncertainty (mu) in qPCR and dPCR measurements for evaluating the performance of quantification methods for nucleic acid target sequences of qPCR and dPCR.
ISO 20395 assists in proper assessment for evaluating and standardizing the performance of quantification methods for nucleic acid target sequences of qPCR and dPCR.
