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Loop-mediated isothermal amplification detection primer group and kit for swine Getah virus

A technology of loop-mediated isothermal and amplified primers, which is applied in the field of zoonotics detection, can solve the problems of inability to detect, and achieve the effects of simple operation, low pollution and simple determination method

Pending Publication Date: 2022-01-18
广东方道基因生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The LAMP method has been developed to detect several viruses such as Japanese encephalitis virus (JEV), foot-and-mouth disease virus, dengue virus, West Nile virus, and Batay virus, but not GETV

Method used

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  • Loop-mediated isothermal amplification detection primer group and kit for swine Getah virus
  • Loop-mediated isothermal amplification detection primer group and kit for swine Getah virus
  • Loop-mediated isothermal amplification detection primer group and kit for swine Getah virus

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Embodiment 1

[0040] 1. Experimental method

[0041] 1 Test strains and bacterial strains

[0042] GETV strain JL17 / 08 was isolated from mosquitoes caught in the wild. The virus proliferated in Vero cells, and the culture supernatant was collected 4 days after infection, and clarified by centrifugation at 12000 g for 30 min. The laboratory isolated and identified Japanese encephalitis virus (JEV, strain YN0901), Akabane virus (AKAV, NM / BS / 1 strain), pseudorabies virus (PRV, DL14 / 08 strain), Batay virus (BATV, NM / 12 strains), porcine reproductive and respiratory syndrome virus (PRRSV, CC-1 strain) or porcine circovirus 2 (PCV2, NM2002 strain) and Sindbis virus (SINV, YN1008 strain).

[0043] 2 standard sample preparation

[0044]Use QIAamp DNA / Virus RNA Mini Kit (QIAAm DNA / Virus RNA Mini Kit) from Vero cell culture supernatant containing 105 plaque-forming units of JL17 / 08, JEV, AKAV, PRV, BATV, PRRSV, PCV2 and SINV RNA was extracted and reverse transcribed into cDNA. The content and p...

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Abstract

The invention discloses a loop-mediated isothermal amplification detection primer group and kit for a swine Getah virus. According to the primer group and the kit, the loop-mediated isothermal amplification primer group for detecting the swine Getah virus is designed according to a conserved region of an nsP1 gene of a genome of the swine Getah virus and is applied to RT-LAMP analysis. A loop-mediated isothermal amplification method for detecting the swine Getah virus is established by using primers designed by the invention, the method is high in specificity, cross reaction with other porcine viruses is avoided, and the lowest detection limit can reach 2.61 copies / microliter. When the primer group and the kit are applied to detection of the swine Getah virus, rapid and instant detection of the virus can be realized, the detection sensitivity and specificity are improved, meanwhile, the labor and equipment cost is reduced, and the period is shortened. The primer group and the kit disclosed by the invention can be popularized and applied to epidemiological investigation and epidemic situation monitoring of the swine Getah virus, and have good practical significance and wide market prospect.

Description

technical field [0001] The invention relates to a detection primer set for ring-mediated isothermal amplification of porcine getavirus, and also relates to a detection kit for ring-mediated isothermal amplification containing the primer set. The invention belongs to the technical field of animal infectious disease detection. Background technique [0002] Porcine Gatah virus (GETV), a mosquito-borne virus, is considered a novel pathogen. GETV is transmitted by mosquitoes, and vertebrates can act as amplifying hosts during seasonal outbreaks. GETV can cause rashes, fever, and swollen legs in horses, abortion and death in pigs, and even fever in humans. [0003] To effectively respond to a possible outbreak, GETV needs to be detected at an early stage, even before symptoms appear, using a method with strong specificity, good accuracy, and high sensitivity. Current diagnostic methods to detect GETV include virus isolation and virus neutralization assay (VN), hemagglutination ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2531/119C12Q2537/1376C12Q2563/107C12Q2521/107
Inventor 刘昊李丽霞黄良宗刘明杰陈盛楠
Owner 广东方道基因生物科技有限公司