SN/T 3492-2013Quarantine protocol for infectious myonecrosis (English PDF)

Foreword

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

This standard reference OIE "aquatic animal disease diagnostic manual (2011 edition)" chapter 2.2.3, select clinical diagnosis of OIE manual Yung and molecular diagnosis on reverse transcription - polymerase chain reaction and real-time quantitative reverse transcription - polymerase chain reaction detection method content.

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

This standard was drafted. People's Republic of China SZCIQ, People's Republic of China Hainan Entry-Exit Inspection and Quarantine Bureau, People's Republic of China Hunan Exit Inspection and Quarantine, Shenzhen Inspection and Quarantine Science Research Institute.

Drafters of this standard. Lanvin lift, Zhengxiao Cong, Shixiu Jie, Huang Ji Hui, Tang Lianfei, Jia Peng, Wang Jinjin, Ye Yi gifted, Ying Wang.

Technical specifications quarantine infectious muscle necrosis

1 Scope

This standard specifies the infectious muscle necrosis virus (Infectiousmyonecrosisvirus, IMNV) reverse transcription - polymerase chain reaction should and real-time fluorescence quantitative reverse transcription - polymerase chain reaction detection method.

This standard applies to clinical symptoms of infectious muscle necrosis (Infectiousmyonecrosis, IMN) quarantine and diagnosis.

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.

Laboratory use specifications and test methods GB/T 6682 Analysis GB/T 18088 Entry and Exit Animal Quarantine sampling

3 Terms and Definitions

The following terms and definitions apply to this document.

Reverse transcription - polymerase chain reaction reversetranscriptionpolymerasechainreaction; RT-PCR RNA under the action of reverse transcriptase, under suitable reaction conditions, is reverse transcribed into cDNA, as a template in a polymerase chain cDNA Reactor.

Real-time quantitative reverse transcription - polymerase chain reaction real-timeRT-PCR on the basis of conventional RT-PCR, adding a specific fluorescent probes, fluorescent PCR instrument monitoring system can receive the fluorescence signal in the PCR reaction, amplification of a DNA strand of each, there is formed a fluorescent molecule, a cumulative fluorescent signal PCR product Forming a fully synchronized.

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

4 Reagents and materials

4.1 Unless otherwise specified, this standard uses only recognized as an analytical reagent.

4.2 Water. should be consistent with GB/T 6682 stipulated a water.

4.3 Taq enzyme. -20 °C to save.

4.4 reverse transcriptase. -20 °C to save.

4.5 RNase inhibitor. -20 °C to save.

4.6 T7RNA polymerase. -20 °C to save.

4.7 DNaseI. -20 °C to save.