SN/T 3661-2013Detection for rotavirus (group A) of foodbrone disease by real-time fluorescence PCR at ports (English PDF)

Foreword

This standard was drafted in accordance with GB/T 1.1-2009 given rules.

This standard is proposed and managed by the National Certification and Accreditation Administration Committee.

This standard was drafted. People's Republic of China Hebei Exit Inspection and Quarantine.

1 Scope

This standard specifies the border crossings foodborne rotavirus (A group) of PCR for detection methods, including specimen collection, transport and Preservation, processing, nucleic acid extraction and detection results of the analysis and the corresponding bio-security measures.

This standard applies to border crossings immigration stool samples of patients with diarrhea, vomit and suspect food rotavirus (A group) screening test.

2 Normative references

The following documents for the application of this document is essential. For dated references, only the dated version suitable for use herein Member. For undated references, the latest edition (including any amendments) applies to this document.

GB/T 4789.1-2003 Microbiological examination of food hygiene - General GB 19489 General requirements for laboratory biosafety SN/T 1720-2006 rotavirus infection monitoring entry and exit points of order

3 Terms and Definitions

The following terms and definitions apply to this document.

Rotavirus

Belonging reoviridae Rotavirus is a major cause mammalian and avian pathogens diarrhea. Rotavirus is a double-stranded RNA viruses, genetic component segments 11, 7 rotavirus serum (AG), wherein A, B, C associated with human disease group, A group Rotavirus The main cause of diarrhea among children, caused mainly group B rotavirus diarrhea in adults.

Fluorescent PCR fluorescencepolymerasechainreaction Also known as real-time PCR (real-timePCR), is the use of fluorescent dyes on the basis of ordinary PCR under the excitation light effect released Fluorescent light changes directly reflect the amount of PCR product changing technology. The standard used was Taq Man hydrolysis probe Law, the principle is the conventional RT-PCR (reverse transcription-polymerase chain reaction) based on the addition of a specific fluorescent probe that for the period of oligonucleotides, both ends of the report marks a fluorophore and a quencher fluorophore probe is intact, the reporter group launched the fluorescent signal is quenched groups absorb, PCR amplification using Taq DNA polymerase 5'-3 'exonuclease activity of the enzyme hydrolysis probe, make fluorescence report Group and a quencher group separation, the fluorescence monitoring system can receive the fluorescent signal, the PCR product accumulation and fluorescence signals Formation sync.

Ct values cyclethreshold the number of cycles when fluorescence signal of each reaction tube reaches a set threshold experienced.

4 Instruments and equipment

4.1 real-time PCR instrument.