GB/T 22287-2008Detection of hepatitis A virus in shellfish by contentional RT-PCR and real-time fluorescence RT-PCR (English PDF)

Foreword

Appendix A of this standard is a normative appendix. This standard is proposed and managed by the National Certification and Accreditation Administration Committee. This standard was drafted. People's Republic of China Beijing Entry-Exit Inspection and Quarantine Bureau, People's Republic of China Jiangsu Entry-Exit Inspection and Quarantine Bureau. The main drafters of this standard. Chan Kwong-chuen, Rao red, para Hong An, Feng Qian, Fu Pu Bo, Zhang Huiyuan, Wang Qi, Zeng Jing, Zhang Rui, Li Jinhua. Detection of hepatitis A virus in shellfish RT-PCR method and ordinary Real-time fluorescent RT-PCR.

1 Scope

GB/T 22287-2008 is the Chinese national standard on detection of hepatitis a virus in shellfish by contentional rt-pcr and real-time fluorescence rt-pcr, in the field of natural and applied sciences. The /T suffix marks it as a recommended standard: it is not compulsory by itself, but it becomes binding as soon as a contract, a tender or a customer specification calls it up - which in practice is how most foreign buyers meet it. It was issued on 12 August 2008 by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China; Standardization Administration of China, and has been in force since 1 December 2008. Classification: ICS 07.100, CCS X04. This page is published from the official record of the standard held by the Chinese standards administration: the identification, the dates, the classification and the issuing body are taken from there. The clause text, the tables and the numeric limits are in the document itself, which is delivered complete in English translation.

This standard specifies the hepatitis A virus in shellfish ordinary fluorescent RT-PCR and RT-PCR detection method. This standard applies to shellfish to detect hepatitis A virus nucleic acid.

2 Normative references

The following documents contain provisions which, through reference in this standard and become the standard terms. For dated references, subsequent Amendments (not including errata content) or revisions do not apply to this standard, however, encourage the parties to the agreement are based on research Whether the latest versions of these documents. For undated reference documents, the latest versions apply to this standard.

GB/T 6682 analytical laboratory use specifications and test methods ( GB/T 6682-2008, ISO 3696. 1987, MOD)

GB 19489 General requirements for laboratory biosafety SN/T 1193 genetic analysis and detection laboratory technical requirements

3 Terms and Definitions

The following terms and definitions apply to this standard.

3.1 DNA template first high temperature denaturation into single strands, at a suitable temperature and buffer, two primers and the template DNA strand two Some complementary sequence on the occurrence of annealing, followed by catalytic DNA polymerase in four dNTP as substrate, annealing of primers to be extended Stretch, and so forth denaturation, annealing and extension that is located between two primer sequences of the DNA fragment was amplified geometrically.

3.2 RNA in the action of reverse transcriptase, under suitable reaction conditions, is reverse transcribed into cDNA, cDNA as template to PCR.

3.3 Real-time fluorescent RT-PCR method is based on the conventional RT-PCR, to add a specific fluorescent probe. The probe is Period of oligonucleotides, both ends of the report marks a fluorophore and a quencher fluorophore. The probe is intact, the fluorescence emitted by the reporter group The optical signal is quencher absorption; PCR amplification, T Rao q enzyme 5'-3 'exonuclease activity of the enzyme degradation of the probe, and to make the report fluorophore Separating quencher fluorophore, fluorescence monitoring system so that the fluorescent signal may be received, i.e., each strand of DNA amplification, there is a Fluorescence Sub formed to achieve a cumulative fluorescent signal PCR product formed completely synchronized.

3.4 When the number of cycles of each reaction tube fluorescent signal reaches the set value of the field experienced.

4 Abbreviations

The following abbreviations apply to this standard.