T/CCAA 79-2024 Waterborne virus detection method: Real-time RT-PCR method

Scope

Scope: This document specifies the real-time fluorescent RT-PCR detection method for stellate viruses in water bodies. This document is applicable to the qualitative detection of stellate viruses in water bodies, including source water, packaged drinking water, water used in food processing, aquaculture water, seawater, sewage from medical institutions, and groundwater, etc. Main technical content: Foreword II1   Scope 12   Normative reference documents 13   Terms and definitions 14   Equipment and material requirements 15   Reagents 26   Operating steps 26.1   Sample collection and processing 26.1.1   Membrane filtration method 26.1.2   Automatic water sampling and microbial enrichment system method 36.2   Virus extraction and purification 36.2.1   Virus lysis 36.2.2   Virus RNA extraction 36.2.3   Virus RNA purification 36.3   Real-time fluorescent RT-PCR detection 36.4   Quality control 36.4.1   Blank control 36.4.2   Negative control 46.4.3   Positive control 46.4.4   Process control virus 46.4.5   External amplification control 47   Results and reporting 47.1   Judgment of test validity 47.2   Result determination 47.3   Result reporting 48   Biosafety measures 5 Appendix A (informative)   Removal of RNase and preparation of RNase-free solution 6 Appendix B (normative)   Real-time fluorescent RT-PCR primers and probes 8 Appendix C (informative)   Real-time fluorescent RT-PCR reaction system and parameters 9 Appendix D (informative)   Process control virus Escherichia coli phage MS2 culture and primers and probes 10 Appendix E (informative)   External amplification control RNA preparation 12 Appendix F (informative)   Automatic water sampling and microbial enrichment system 13 References

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