GB/T 42077-2022 Active National standards

GB/T 42077-2022 Biotechnology—Requirements for evaluating the performance of quantification methods for nucleic acid target sequences—qPCR and dPCR

Publish Date: 2022-12-30 Implement Date: 2022-12-30 For services related to genuine standard inquiry, procurement, translation, and other related services in China, please Contact Us

Basic Information

Standard Code: GB/T 42077-2022

Standard Type: National standards

Standard Status: Active

is_force_gb: no

CCS Name: Basic disciplines

ICS Name: Biology, botany, and zoology

Publish Date: 2022-12-30

Implement Date: 2022-12-30

Pages: 46 pages

Scope

This document specifies the general requirements for the performance evaluation and quality assurance of nucleic acid target sequence quantification methods. This document is applicable to the quantitative determination of DNA and RNA target sequences using digital PCR (dPCR) or real-time quantitative PCR (qPCR) amplification techniques. Applicable target sequence nucleic acid molecules include 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) and messenger RNA (mRNA); and double-stranded RNA, including long and short non-coding RNAs (such as microRNA (miRNA) and small interfering RNA (siRNA)). This document is applicable to nucleic acids from biological sources (such as viruses, prokaryotic cells, and eukaryotic cells), non-cellular biological fluid samples (such as plasma or cell matrix), and nucleic acids extracted from in vitro non-biological sources (such as oligonucleotides, synthesized gene fragments, and in vitro transcribed (IVT) RNA). This document does not apply to the quantification of extremely short DNA oligonucleotides (<50 bases). This document includes: -- Analysis design, including quantification strategies (qPCR standard curve method, dPCR molecular counting method, qPCR relative quantification method, dPCR ratio quantification method) and use controls; -- Total nucleic acid quality concentration quantification and nucleic acid sample quality control, including nucleic acid quality evaluation (purity and integrity); -- PCR experiment design, optimization, bioinformatics, and in vitro specificity testing; -- Data quality control and analysis, including acceptance criteria, threshold setting, and normalization methods; -- Method validation based on specific qPCR/dPCR requirements (precision, linearity, quantification limit, detection limit, accuracy, and robustness); -- Methods for establishing metrological tr