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SN/T 4275-2015: Detection of Burkholderia pseudomallei by real-time PCR at frontier ports
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SN/T 4275-2015: Detection of Burkholderia pseudomallei by real-time PCR at frontier ports

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Detection of Burkholderia pseudomallei by real-time PCR at frontier ports People's Republic of China Entry-Exit Inspection and Quarantine Industry Standard Real-time port Becker's disease in the border port Fluorescent PCR detection method Released on.2015-05-26 2016-01-01 implementation People's Republic The General Administration of Quality Supervision, Inspection and Quarantine issued

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard is proposed and managed by the National Certification and Accreditation Administration. This standard was drafted. Sichuan Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China, Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Shanghai Zhijiang Biotechnology Co., Ltd. The main drafters of this standard. Tian Lubo, Fan Xuejun, Yan Wendong, Chen Xiaoyu, Shi Ying, Yang Yu, Ni Weiqin, Zhu Xuping, Wang Ning. Real-time port Becker's disease in the border port Fluorescent PCR detection method

1 Scope

This standard stipulates the target of fluorescent PCR detection of Burkholderia sinensis in border ports, collection, transportation and preservation of specimens. Program and results report. This standard is applicable to the screening and detection of Burkholderia sinensis in border ports.

2 Normative references

The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this article. Pieces. For undated references, the latest edition (including all amendments) applies to this document. GB/T 6682 Analytical laboratory water specifications and test methods GB 19489 General requirements for laboratory biosafety

3 Terms and definitions

The following terms and definitions apply to this document. 3.1 Bogholderiapseudomallei It is a Gram-negative bacteria and is a pathogen causing snot of severe infectious diseases in humans and animals. The strain is widely distributed in Southeast Asia and Australia. In the water and soil of northern Italy. China's epidemic areas are mainly distributed in Hainan, Guangdong, Guangxi, Hunan, Guizhou, Fujian, Hong Kong, Taiwan and other provinces. Area. The population is generally susceptible to the bacteria, and the clinical symptoms are complex and diverse, and are classified into acute sepsis, subacute, chronic and subclinical. The most serious symptoms are pneumonia and acute sepsis. The disease has a high mortality rate. 3.2 Fluorescent PCR fluorescencepolymerasechainreaction Also known as real-time PCR (real-time PCR), there are many real-time fluorescent PCR methods, this standard uses TaqMan hydrolysis probe The principle of the needle method is to add a specific fluorescent probe based on conventional PCR, the probe is an oligonucleotide, and the two sides are respectively A fluorescent reporter group and a fluorescent quenching group are labeled. When the probe is intact, the fluorescent signal emitted by the reporter group is absorbed by the quenching group. During PCR amplification, the probe is hydrolyzed by the 5'-3' exonuclease activity of the Taq enzyme to separate the fluorescent reporter group from the fluorescence quenching group. Thereby the fluorescence monitoring system can receive the fluorescent signal. Every time a PCR cycle is passed, the fluorescent signal is the same as the target segment. In the process of synchronous exponential growth, the intensity of the signal is proportional to the nucleic acid content of the detected object, and the detection object can be calculated according to the mathematical model. content. 3.3 Ct value cyclethreshold That is, the cycle threshold refers to the number of cycles experienced when the detected fluorescent signal in each reaction tube just reaches the set threshold.

4 Abbreviations

The following abbreviations apply to this document. ...