GB/T 42077-2022
Biotechnology—Requirements for evaluating the performance of quantification methods for nucleic acid target sequences—qPCR and dPCR (English Version)
- Standard No.
- GB/T 42077-2022
- Language
- Chinese, Available in English version
- Release Date
- 2022
- Published By
- General Administration of Quality Supervision, Inspection and Quarantine of the People‘s Republic of China
- Latest
- GB/T 42077-2022
- Scope
- This document specifies general requirements for the performance evaluation and quality assurance of methods for the quantification of nucleic acid target sequences. This document is applicable to the quantitative determination of DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) target sequences by 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 (s sDNA), complementary DNA (cDNA), single-stranded RNA (s sRNA) including ribosomal gDNA) and plasmid DNA; single-stranded DNA (RNA (rRNA), messenger RNA (mRNA); double-stranded RNA, including long and short non-coding RNA [microRNA (miRNA) and small interfering RNA (siRNA), etc.]. This document applies to nucleic acids of biological origin (such as viruses, prokaryotic cells, and eukaryotic cells), cell-free biological fluid samples (such as plasma or cell matrix); Extracted nucleic acids. This document is not suitable for the quantification of DNA oligonucleotides that are too short (<50 bases). This document contains: - Assay design, including quantification strategies (dPCR molecular counting method, dPCR versus qPCR standard curve Quantification of total nucleic acid mass concentration and quality control of nucleic acid samples, including assessment of nucleic acid quality (purity and integrity); ——PCR test design, optimization, Bioinformatics and in vitro specificity testing; - data quality control and analysis, including acceptance criteria, threshold setting and normalization methods; - method validation (precision, linearity, limit of quantitation) according to specific requirements of qPCR/dPCR , detection limit, correctness and robustness); ——methods for establishment of metrological traceability and assessment of measurement uncertainty. This document does not cover the sampling of biological materials or the processing of biological samples (such as collection, preservation, transportation, storage , processing and nucleic acid extraction) put forward requirements or acceptance criteria. Nor does it put forward requirements and acceptance criteria for special applications (such as food or clinical application areas where special matrix problems may occur).
GB/T 42077-2022 Referenced Document
GB/T 42077-2022 history
- 2022 GB/T 42077-2022 Biotechnology—Requirements for evaluating the performance of quantification methods for nucleic acid target sequences—qPCR and dPCR
