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Primers for detection of porcine Japanese encephalitis virus and visual kit

A Japanese encephalitis virus, epidemic technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of insufficient detection efficiency, unfavorable promotion, and the need for expensive equipment To achieve the effect of high sensitivity and specificity, simple detection procedure, and optimized reaction system

Inactive Publication Date: 2013-05-15
INSPECTION & QUARANTINE TECH CENT OF GUANGDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] However, the above-mentioned prior art still has shortcomings and deficiencies: ① the judgment of the experimental results is not fast and intuitive enough; ② expensive instruments and equipment are needed, which is not conducive to popularization; ③ the detection efficiency is not high enough

Method used

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  • Primers for detection of porcine Japanese encephalitis virus and visual kit
  • Primers for detection of porcine Japanese encephalitis virus and visual kit
  • Primers for detection of porcine Japanese encephalitis virus and visual kit

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Experimental program
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Effect test

Embodiment Construction

[0061] 1. LAMP amplification of the conserved region of the JEV NS3 gene:

[0062] Taking JEV total RNA as a template, using the primers provided by the invention to carry out the amplification of the conserved region of the JEV NS3 gene, the reaction system is:

[0063] The 25μL reaction system contains:

[0064] 10×Thermopol reaction buffer (NEB company), 2.5 μL;

[0065] JEV total RNA 5.25μL;

[0066] 8U / tube Bst DNA polymerase large fragment (NEB company), 1 μL;

[0067] AMV reverse transcriptase (NEB company) 0.25 μL;

[0068] 10mmol / L dNTPs, 2.5μL;

[0069] 20pmol / tube upstream internal primer (FIP), 0.8μL;

[0070] 20pmol / tube downstream internal primer (BIP), 0.8μL;

[0071] 5pmol / tube upstream external primer (F3), 0.2μL;

[0072] 5pmol / tube downstream external primer (B3), 0.2μL;

[0073] 20pmol / tube upstream loop primer (FL), 0.4μL;

[0074] 20pmol / tube downstream loop primer (BL), 0.4μL;

[0075] (The above primers were synthesized by Bao Biological Enginee...

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Abstract

The invention provides a set of primers for detection of a porcine Japanese encephalitis virus. The primers include: an upstream internal primer, a downstream external primer, an upstream external primer, a downstream external primer, an upstream loop primer and a downstream loop primer. The sequences of the primers are shown as SEQ ID No.1-6 in a sequence table. The invention also provides a visual kit that contains the set of primers and is used for detecting the porcine Japanese encephalitis virus and a method for detecting whether a non-living sample contains the porcine Japanese encephalitis virus by the kit. The primers, the kit and the detection method provided in the invention optimizes the reaction system, and make the result determination realize visualization, thus being of great significance in detection, diagnosis, prevention and treatment of porcine Japanese encephalitis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, to a rapid detection kit and primers for swine infection with Japanese encephalitis virus. Background technique [0002] Japanese encephalitis virus (JEV), also known as Japanese encephalitis virus (JEV), is referred to as Japanese encephalitis virus, and the disease it causes is referred to as Japanese encephalitis. Humans and a variety of animals can cause central nervous system diseases in humans and reproductive disorders in pigs, which seriously endanger human health and the development of animal husbandry. [0003] The laboratory diagnostic method of encephalitis E etiology can detect viral antigens in the brain tissue, fetus, placenta, and body fluids of cases. The current diagnostic methods are: immunocytochemistry, reverse passive hemagglutination test, fluorescent antibody staining and RT-PCR technology. Using the traditional method of gene amplification, Nagasaki Universit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 林志雄田纯见何晓明罗琼鱼海琼罗长保陈茹刘志玲王宏吴晓薇毕英佐马静云
Owner INSPECTION & QUARANTINE TECH CENT OF GUANGDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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