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Universal RPA (recombinase polymerase amplification) nucleic acid isothermal amplification primer and kit for herpesvirus abalone HaHV-1 and application of universal RPA nucleic acid isothermal amplification primer

A herpes virus, isothermal amplification technology, applied in the direction of DNA / RNA fragments, biological material analysis, recombinant DNA technology, etc., can solve the problem that it is difficult to meet the needs of rapid on-site detection of epidemic diseases, and achieve good and good specificity test results The effect of stability and accurate detection results

Pending Publication Date: 2022-03-01
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

Moreover, the existing methods are difficult to meet the needs of front-line farmers for on-site rapid detection of epidemic diseases in the production process

Method used

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  • Universal RPA (recombinase polymerase amplification) nucleic acid isothermal amplification primer and kit for herpesvirus abalone HaHV-1 and application of universal RPA nucleic acid isothermal amplification primer
  • Universal RPA (recombinase polymerase amplification) nucleic acid isothermal amplification primer and kit for herpesvirus abalone HaHV-1 and application of universal RPA nucleic acid isothermal amplification primer
  • Universal RPA (recombinase polymerase amplification) nucleic acid isothermal amplification primer and kit for herpesvirus abalone HaHV-1 and application of universal RPA nucleic acid isothermal amplification primer

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Effect test

Embodiment 1

[0037] Example 1: Establishment of the test strip RPA detection method for rapid on-site detection of HaHV-1

[0038] 1. Design of RPA upstream and downstream primers and probe primers

[0039] The genome sequences of HaHV-1 mutant strains KU096999.1 and JX453331.1 were downloaded from GenBank of NCBI and compared and analyzed, and the conserved segment among HaHV-1 mutant strains was screened out, and a set of primers was designed for the conserved segment, in order to HaHV-1 of different mutant strains can be detected. The present invention designed and synthesized five sets of primers respectively, as shown in Table 1 below.

[0040] Table 1

[0041]

[0042] 2. DNA extraction from clinical samples

[0043]All clinical HaHV-1 positive samples used in the present invention are collected and preserved by our laboratory. The TIANamp Marine Animals DNA Kit from TIANGEN was used to extract the total DNA of the mantle of the abalone according to the instructions, and final...

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Abstract

The invention belongs to the technical field of molecular biological detection, and particularly relates to a herpes abalone virus HaHV-1 universal RPA nucleic acid isothermal amplification primer, a kit and application thereof. The primer is a complete set of primer combination and consists of upstream and downstream primers and a probe primer, the upstream primer is as shown in SEQ ID NO. 1; the downstream primer is as shown in SEQ ID NO. 2; and the probe primer is as shown in SEQ ID NO. 3. According to the present invention, the universal RPA amplification detection primer is designed according to the conservative segment aiming at the different variants of the existing known herpes abalone virus HaHV-1, the test strip RPA detection system and the test strip RPA detection method of the HaHV-1 are established, and the technology has important values on the rapid on-site screening and detection of the HaHV-1 involved in the breeding process or the import and export trade, and has broad market application prospects.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to an abalone herpes virus HaHV-1 universal RPA nucleic acid isothermal amplification primer, a kit and an application thereof. Background technique [0002] Abalone virus disease and abalone cracking shell disease are listed as three types of animal diseases in my country's "List of Types I, II, and III Animal Epidemic Diseases" and listed as Type II imported animal diseases in the "List of Imported Animal Quarantine Epidemic Diseases of the People's Republic of China" . This kind of disease seriously threatens the farming of abalone in the north and south of our country. Since 1999, in southern China, mainly in Dongshan District, Fujian Province, abalone herpes virus has caused large-scale deaths of abalone. Then, in 2003 and 2005, variegated abalone and Australian black lip abalone (Haliotis rubra ), green-lipped abalone (Haliotis laevigata) and i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11G01N33/543G01N33/558
CPCC12Q1/705C12Q1/6844G01N33/54313G01N33/558G01N2333/03C12Q2521/507C12Q2522/101
Inventor 辛鲁生于江南白昌明王崇明
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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