TaqMan-MGB fluorescent quantitative polymerase chain reaction (PCR) detection primer, TaqMan-MGB fluorescent quantitative PCR detection probe and TaqMan-MGB fluorescent quantitative PCR detection method for seneca valley virus (SVV)

Abstract

The invention relates to the technical field of molecular biology detection, in particular to a TaqMan-MGB fluorescent quantitative polymerase chain reaction (PCR) detection specific primer (as shown in SEQ ID No. 1 and SEQ ID No. 2), a TaqMan-MGB fluorescent quantitative PCR detection probe as shown in SEQ ID No. 3) and a TaqMan-MGB fluorescent quantitative PCR detection method for seneca valley virus (SVV). The method comprises the steps of drawing a standard curve; extracting ribonucleic acid (RNA) of a sample virus; carrying out reverse transcription on the RNA of the sample virus; enabling the product of the reverse transcription to have a TaqMan-MGB fluorescent quantitative PCR, and reading the result. The primer provided by the invention has better specificity and sensitivity; the MGB probe provided by the invention is shorter and is beneficial to probe design, and the Tm value difference between a paired template and a non-paired template is improved, so that the experimental result is more stable and accurate; the method provided by the invention has the advantages of being simple and rapid, easy to operate, visual in results, high in sensitivity, good in stability, real-time quantitative, and the like, and shortens the reaction time.

Description

technical field [0001] The invention relates to a detection method in the technical field of molecular biology detection, in particular to a specific primer, probe and detection method for Seneca Valley virus TaqMan-MGB fluorescence quantitative PCR detection. Background technique [0002] Seneca Valley virus (SVV) is a single-stranded positive-sense RNA virus belonging to the genus Senecavirus of the picornaviridae family. SVV was first discovered in PER.C6 cell cultures in 2002. In 2007, SVV was identified as the causative agent of blister symptoms in pigs in a group of slaughterhouses in the United States. No new reports appeared until 2015. Brazil and Many SVV outbreaks have broken out in the United States, and pigs at multiple stages can be infected, and blisters appear on the nose, lips, and hoofs, and lead to the death of young piglets, causing greater losses. Subsequently, there were reports that Seneca Valley virus also appeared in China. Since it had never appeare...

AI Technical Summary

Problems solved by technology

SVV was first discovered in PER.C6 cell cultures in 2002. In 2007, SVV was identified as the causative agent of blister symptoms in pigs in a group of slaughterhouses in the United States. No new reports appeared until 2015. Brazil and Many SVV outbreaks have broken out in the United States, and pigs at multiple stages can be infected. Blisters appear on the nose, lips and hooves, and lead to the death of young piglets, causing greater losses.

Subsequently, there were reports that Seneca Valley virus also appeared in China. Since it had never appeared in China before, there was no ready-made detection method. Up to now, there are still few detection methods for Seneca Valley virus in China. The main reason is that the disease is a new disease. The existing RT-PCR detection method can only perform qualitative detection but not accurate quantitative analysis, and electrophoresis is required to judge the results after amplification. The number of samples tested each time is small, and the samples with low content Disadvantages such as insufficient resolution

Images