RT-LAMP nucleic acid test-strip kit for determining hog cholera virus and application

Abstract

The invention discloses a RT-LAMP nucleic acid test-strip kit for determining hog cholera virus and application. The kit comprises a primer group with nucleotide sequences shown as SEQ ID NO. 1-6 and nucleic acid detection test strips. The application method of the kit comprises as follows: preparing a RT-LAMP reaction system which comprises AMV retrovirus, a 1* reaction buffer, strand displacement DNA polymerase, a dNTP mixture, betaine, MgSO4, a FIP primer, a BIP primer, a hybridization probe, a LoopB primer, a F3 primer, a B3 primer and RNA of a sample to be measured; and carrying out a reaction at a constant temperature, after testing the obtained products by using the nucleic acid detecting test strip, judging and reading directly: the result is positive when two red bands appear, and one band is in the detection zone while the other band is in the control zone. The kit has the advantages of simple operation, low cost, easy observation of the reaction result, good specificity and easy popularization and application in large scope.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an RT-LAMP nucleic acid test strip kit for detecting swine fever virus and its application. Background technique [0002] Classical swine fever (CSF) is a severe infectious disease with high morbidity and mortality caused by classical swine fever virus (CSFV). The infection process can be divided into acute, subacute, chronic, Delayed and so on. Acute cases of CSF are more common in the early stage of the epidemic. The main manifestations are sudden onset, cyanosis of the skin and mucous membranes, and general convulsions; constipation at first, and diarrhea later. In the later period, he appeared unsteady in gait, then became paralyzed, fell to the ground and died. Less virulent CSFV strains can cause subacute CSF or mild CSF with milder symptoms. After infection of pregnant sows, "poisoned sow syndrome" occurs, miscarriage, stillbirth, weak fetus, mummified fetus, death of newbo...

AI Technical Summary

Problems solved by technology

The reaction result can be judged by visual observation, or by measuring the turbidity of the white precipitate formed by the by-product magnesium pyrophosphate obtained during the reaction with a turbidimeter, but the turbidimeter is expensive and inconvenient to carry

Some scholars use the method of adding SYBR Green I dye to the reaction system to detect the amplification product, but this observation requires uncapping treatment. The high efficiency and high sensitivity of LAMP amplification make the product contain a large number of target fragments. It is extremely easy to pollute the environment by adding dyes

Therefore, it was invented to add fluorescent dyeing reagents into the reaction tube, observe under the ultraviolet light, monitor the reaction product in real time, and realize the visualization of LAMP, but this method is not accurate enough for the judgment of weak positive

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