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PCR primer group for rapidly detecting three fish viruses and application thereof

A primer set and virus technology, applied in the field of gene detection, can solve the problems of easy cross-infection, time-consuming and labor-intensive, and achieve the effects of high sensitivity, improved efficiency and strong specificity

Active Publication Date: 2021-02-12
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, many studies have established single and dual PCR detection methods for ISKNV, SCRIV and SCRV, but these conventional detection methods can only detect 1-2 virus types at a time, and there are disadvantages such as prone to cross-infection, time-consuming and labor-intensive

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  • PCR primer group for rapidly detecting three fish viruses and application thereof
  • PCR primer group for rapidly detecting three fish viruses and application thereof
  • PCR primer group for rapidly detecting three fish viruses and application thereof

Examples

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Embodiment 1

[0043] Example 1 Establishment of Triple PCR Detection Method for Infectious Spleen and Kidney Necrosis Virus, Mandarin Fish Frog Virus and Mandarin Rhizoma Virus

[0044] 1.1 Design and synthesis of primers

[0045] According to the MCP gene sequence of ISKNV (HQ317460.1) and its conserved region in GenBank, primers were designed respectively: ISKNV-MCP-F / ISKNV-MCP-R; the MCP gene sequence of SCRIV (MG941005.1) and its conserved region were designed respectively : SCRIV-MCP-F / SCRIV-MCP-R; SCRV N gene sequence (NC008514.1) and its conserved region were designed primers: SCRV-N-F / SCRV-N-R.

[0046] The primers were synthesized by Guangzhou Aiji Biotechnology Co., Ltd., and the primer information is shown in Table 1.

[0047] Table 1

[0048]

[0049] 1.2 Construction of standard recombinant plasmids

[0050] Using the DNA of clinical samples infected with ISKNV as a template, and using ISKNV-MCP-F / ISKNV-MCP-R as primers to amplify the MCP gene of ISKNV by PCR, the fragmen...

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Abstract

The invention discloses a triple PCR primer group for rapidly detecting three fish viruses and application thereof, and relates to the technical field of gene detection. The invention discloses a triple PCR primer group for rapidly detecting three fish viruses. The triple PCR primer group comprises an infectious spleen and kidney necrosis virus ISKNV primer, a mandarin fish frog virus SCRIV primerand a mandarin fish rhabdovirus SCRV primer, the triple PCR primer group has high sensitivity, the detection time is 2-3 hours earlier than that of conventional PCR, the efficiency of clinical diagnosis is greatly improved, richer reference bases can be provided for epidemiological investigation of ISKNV, SCRIV and SCRV infection, and the triple PCR primer group has important significance for subsequent research and prevention and treatment.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a PCR primer set for rapid detection of three fish viruses and its application. Background technique [0002] Mandarin fish has a long history of breeding and is one of the high-quality freshwater fish in my country. It is loved by consumers for its high yield, tender meat, less spines, sweet taste, and rich protein. However, with the rapid expansion and development of mandarin fish farming technology, equipment and feed market in China, Infectious spleen and kidney necrosis virus (ISKNV), mandarin fish frog virus (Siniperca chuatsi ranairidovirus, SCRIV) and mandarin shellfish Viral diseases such as Siniperca chuatsi rhabdovirus (SCRV) have become the main factors restricting the sustainable development of mandarin fish aquaculture. ISKNV is one of the main pathogens of large-scale outbreaks of infectious diseases in mandarin fish, and its clinical symptoms usually manif...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2537/143Y02A50/30
Inventor 梁红茹马赛亚付小哲李宁求林强刘礼辉牛银杰罗霞黄志斌
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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