T/CCAA 78-2024 Detection of Hepatitis A Virus in Water Using Real-Time Fluorescent RT-PCR Method

Scope

Scope: This document specifies the real-time fluorescent RT-PCR method for detecting the hepatitis A virus in water bodies. This document is applicable to the qualitative detection of the hepatitis A virus 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 26.2   Virus extraction and purification 36.2.1   Virus lysis 36.2.2   Viral RNA extraction 36.2.3   Viral RNA purification 36.3   Real-time fluorescent RT-PCR detection 36.4   Quality control 36.4.1   Blank control 36.4.2   Negative control 36.4.3   Positive control 36.4.4   Process control viruses 46.4.5   External amplification control 47   Results and reporting 47.1   Determination of test validity 47.2   Result determination 47.3   Result reporting 48   Biosafety measures 4 Appendix A (informative)   Removal of RNase and preparation of RNase-free solution 5 Appendix B (normative)   Real-time fluorescent RT-PCR primers and probes 7 Appendix C (informative)   Real-time fluorescent RT-PCR reaction system and parameters 8 Appendix D (informative)   Process control viruses Escherichia coli phage MS2 culture and primers and probes 9 Appendix E (informative)   External amplification control RNA preparation 11 Appendix F (informative)   Automatic water sampling and microbial enrichment system 12 References 15

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