Type A seneca virus detection primer, kit, virus detection method and application

Abstract

The invention belongs to the technical field of animal pathogen molecule diagnosis, and particularly relates to a type A seneca virus detection primer, a kit, a virus detection method and application.The following primers including an SVA forward primer being SEQ ID NO.1:5'-AGAATTTGGAAGCCATGC-3' and an SVA reverse primer SEQ ID NO.2:5'-GAAACAGAYTGCAGCTTCTC-3' are comprised, wherein the Y represents a degenerate base group, and is a base group T or C. The high sensitivity and specificity are ensured; meanwhile, the detection accuracy is improved; the false positive and false negative probability is reduced.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis of animal pathogens, and in particular relates to a type A Seneca virus detection primer, a kit, a virus detection method and an application. Background technique [0002] Type A Seneca virus (Senecavirus A, SVA), formerly known as Seneca Valley virus (Seneca Valleyvirus), is a new pig disease pathogen in China in recent years, similar to foot-and-mouth disease virus (FMDV) and porcine vesicular disease virus (SVDV) and vesicular stomatitis virus (VSV) are similar, mainly cause the mucosal disease of pigs, and easily cause the death of piglets and the reduction of production of fat pigs, and the economic harm to pig breeding industry is relatively large. [0003] SVA is the only virus in the Senecavirus genus with only one serotype. However, its epidemiology, infection mechanism, and host type are still lacking in international verification. At present, it is generally believed that S...

AI Technical Summary

Problems solved by technology

[0010] 1. Virus isolation and identification has always been the "gold standard" for laboratory diagnosis. The isolated virus can facilitate a series of follow-up related research, but it is difficult for early diagnosis

The detection period of more than 1 week is obviously not suitable for the detection of animal groups, and many viruses cannot be cultured and isolated through existing cell lines, which also limits the versatility of this method

[0011] 2. Due to the existence of the "gap period", it is not suitable to indirectly diagnose SVA infection through antibody detection. In many cases, the pigs are obviously sick, but the antibody still lags behind

Direct detection of viral antigens can be used for early diagnosis, but the sensitivity of antigen detection must be further improved through chemiluminescence and other technologies. Technical barriers and additional expensive equipment investment limit the promotion of on-site detection

[0012] 3. The RT-PCR detection for SVA nucleic acid has the advantage of the highest sensitivity, and has the advantage of "empty window period", which can realize the investigation in the early stage of the disease, but the traditional RT-PCR detection operation still requires high requirements, which is easy to cause false positives Pollution affects the accuracy of diagnosis, and it is difficult to achieve on-site detection

The MGB group has the effect of increasing the Tm value of the probe, reducing the synthesis cost and increasing the success rate, but the probe sequence using this label is often very short (generally less than 20bp), and is often used for the detection of single nucleotide polymorphism SNP. It can distinguish the mutation of 1 base, so it is used for the detection of viruses whose genome is easy to mutate, and often misses detection due to base mutation

[0014] The detection method of Seneca virus type A disclosed in patent CN201710327691 divides RNA reverse transcription and cDNA amplification into two independent steps, and additionally needs to open the cover to transfer samples and prepare reagents in the middle, which not only easily causes operational errors, but also costs more It takes a long time, and it is also easy to cause pollution and affect the accuracy of the test results

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