Pretreatment reagent for nucleic acid extraction of pig farm environment sample as well as preparation method and application thereof

Abstract

The invention discloses a pig farm environment sample nucleic acid extraction pretreatment reagent and a preparation method and application thereof, and relates to the technical field of livestock breeding pathogen detection. The reagent is prepared from the following raw materials in percentage by volume and mass: 0.3 to 0.8 percent of an emulsifier T-20, 0.1 to 0.2 percent of sodium lauryl sulfate, 0.7 to 1 percent of sodium chloride, 1 to 2 percent of ethylenediamine tetraacetic acid, 0.5 to 1 percent of a buffer solution and the balance of water. Impurity proteins and other impurities in pig farm environment samples can be effectively removed, the quality of nucleic acid extraction purity is remarkably improved, and the method plays an important role in efficient settling separation of impurities and high-purity separation and purification of nucleic acid, so that the requirement of a fluorescent quantitative PCR experiment on nucleic acid purity is met, the accuracy of a detection result is effectively improved, and the detection cost is reduced. The problem that impurities affect the detection result is solved, and healthy development of the pig raising industry is guaranteed.

Description

technical field [0001] The invention relates to the technical field of animal husbandry pathogen detection, in particular to a pretreatment reagent for nucleic acid extraction from environmental samples of pig farms and a preparation method and application thereof. Background technique [0002] Fluorescence quantitative PCR detection technology, as a detection method integrating conventional PCR, fluorescent labeling, laser irradiation and digital imaging technology, has the advantages of strong specificity, high sensitivity and low false positive, so it is widely used in the detection of pathogens in animal husbandry, especially It is a pig industry plagued by African swine fever. [0003] However, in the production process of pig herds, there is a problem that the nucleic acid extraction effect of environmental samples is poor, resulting in a low detection rate or false positives. The specific performance is that the composition of pig farm environmental samples is comple...

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Problems solved by technology

[0003] However, in the process of pig production, there is a problem of poor nucleic acid extraction from environmental samples, resulting in low detection rate or false positives.

The specific performance is that the composition of the pig farm environment sample is complex, containing at least but not limited to feed, feces, dust, dirt, knitted fabrics, disinfectants and other impurities, which have a great inhibitory effect on the quality of nucleic acid extraction and purification, thus affecting downstream fluorescence quantification The accuracy of PCR detection results is not conducive to the timely diagnosis of various diseases in pig herds, and is extremely harmful to the healthy development of the pig industry

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