Reagent for detecting SNP locus genotype related to grouper nervous necrosis virus resistance

A nerve necrosis and detection kit technology is applied in the fields of molecular biology and genetic breeding, which can solve the problems of lack of detection of grouper nerve necrosis virus resistance, economic loss, and great harm, and achieves the improvement of anti-nerve necrosis symptoms. ability, speeding up the cultivation process, the effect of speeding up the cultivation process

Active Publication Date: 2021-03-02
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]Red-spotted grouper nervous necrosis virus (RGNNV) is very harmful to adults and juveniles of grouper, serious When the mortality rate can reach 100%, it often causes huge economic losses to the grouper aquaculture and seriously threatens the healthy development of the grouper aquaculture industry. However, there is still no effective treatment for the disease caused by the virus at present. monitoring
[0004] Chinese patent CN201610255794.5 discloses a method for detecting grouper neuronecrosis virus infection based on nucleic acid aptamer-based Sandwich ELASA; CN201610465941.1 discloses a Colloidal gold immunochromatographic detection test paper for detecting seven-banded grouper neuronecrosis virus antibody and its application; CN201910873715.0 discloses a colloidal gold test strip for detecting grouper neuronecrosis virus and its preparation and detection method ; CN200910039534.4 discloses a primer set, detection method and rapid diagnostic kit for detection of red-spotted grouper neuronecrosis virus; CN03114369.5 discloses grouper viral neuronecrosis virus gene diagnostic kit and detection method, However, there is a lack of a method for detecting grouper neuronecrosis virus resistance

Method used

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  • Reagent for detecting SNP locus genotype related to grouper nervous necrosis virus resistance
  • Reagent for detecting SNP locus genotype related to grouper nervous necrosis virus resistance
  • Reagent for detecting SNP locus genotype related to grouper nervous necrosis virus resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0068] Example 2 Confirmation of the SNPs of the PPAR-δ gene of the red-spotted grouper

[0069] 1. Experimental method

[0070] According to the 8 candidate PPAR-δ gene SNP sites obtained in Example 1 and the sequences near them, 50 susceptible samples and 50 anti-sensitivity samples randomly selected were used as templates, and the SnapShot method was used to analyze the SNPs obtained in Example 1. Genotyping was carried out, and the correlation analysis was carried out between the typing results and the resistance to neuronecrosis virus.

[0071] The SnapShot method detection was completed by TsingKe Company, and its amplification primer information and single-base extension primer information are shown in Table 1 and Table 2, respectively. Correlation analysis of resistance to neuronecrosis virus The chi-square test in SPSS 17.0 statistical software was used to conduct genotyping of 8 SNPs and correlation analysis with resistance to neuronecrosis virus (P<0.05 indicated s...

Embodiment 3

[0081] Embodiment 3 A kind of detection method of red-spotted grouper to neuronecrosis virus disease-resistant character

[0082] By detecting the SNP markers of the red-spotted grouper to be tested, the resistance and susceptibility of the red-spotted grouper to be tested to the nerve necrosis virus is determined. Include the following steps:

[0083] (1) extract the genomic DNA of the red-spotted grouper to be tested;

[0084] (2) using the DNA obtained in step (1) as a template, and using the nucleotide sequences shown in SEQ ID NO.2 to 3 as primers, to perform PCR amplification to obtain PCR amplification products,

[0085] SEQ ID NO. 2: 5'-gtgggaggaatgctgatgtg-3'.

[0086] SEQ ID NO. 3: 5'-attcttcttcaccgtggaggc-3'. ;

[0087] (3) the PCR amplification product obtained in step (2) is sequenced to determine the genotype of the SNP marker of the red-spotted grouper to be tested;

[0088] (4) Determine whether the red-spotted grouper to be tested is resistant to neuronec...

Embodiment 4

[0090] Example 4 A kit for detecting the disease-resistant traits of red-spotted grouper to neuronecrosis virus

[0091] 1. Composition

[0092] Primers and PCR amplification reagents with nucleotide sequences as shown in SEQ ID NO.2-3;

[0093] Wherein, SEQ ID NO.2: 5'-gtgggaggaatgctgatgtg-3';

[0094] SEQ ID NO. 3: 5'-attcttcttcaccgtggaggc-3'.

[0095] 2. How to use

[0096] With embodiment 3.

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Abstract

The invention discloses a reagent for detecting SNP locus genotype related to grouper nervous necrosis virus resistance. The invention discloses an SNP (Single Nucleotide Polymorphism) site related tonervous necrosis virus resistance. The death probability of groupers with the SNP site genotype of CT (Computed Tomography) infected with nervous necrosis viruses is obviously lower than that of individuals with the genotype of CC or TT. By detecting the genotype of the SNP of the epinephelus akaara, whether the epinephelus akaara has nervous necrosis virus resistance can be effectively determined. The reagent is advantaged in that the SNP marker provided by the invention is closely related to the resistance of epinephelus akaara to nervous necrosis viruses, selection of individuals with CT genotypes in parent breeding is beneficial to improving the nervous necrosis disease resistance of offspring, and the marker provided by the invention is used for auxiliary breeding, so the culture process of excellent disease-resistant varieties of epinephelus akaara can be accelerated.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and genetic breeding, in particular to a reagent for detecting the genotype of a SNP site related to grouper neuronecrosis virus resistance. Background technique [0002] Red-spotted grouper (Epinephelus akaara), commonly known as red spot, belongs to the genus Epinephelus of the family Serranidae in the order Perciformes. It is mainly distributed in the western part of the North Pacific Ocean. It is produced in the south of the Zhoushan Islands in China and through the Taiwan Strait to the South China Sea. , Beibu Gulf area. It has the characteristics of delicious meat and high nutritional value, and is an important grouper breeding species in the coastal areas of South China. [0003] Red-spotted grouper nerve necrosis virus (red-spotted grouper nervous necrosis virus, RGNNV) is very harmful to adults and juveniles of grouper, and the mortality rate can reach 100% in severe cases, of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 王庆秦启伟杨敏陈锦鹏黄健玲李鑫帅郑乐云
Owner SOUTH CHINA AGRI UNIV
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