Specific primer for detecting Aleutian disease virus of minks and application of specific primer in screening healthy minks

A technology of mink Aleutian and Aleutian virus, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of tedious sampling, inconvenience, hindering mink breeding work, etc., and achieve experimental stability. High performance, high coincidence rate, stable application effect

Inactive Publication Date: 2015-03-25
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned methods are cumbersome and inconvenient in sampling to varying degrees, and also hinder the breeding work of mink. Therefore, it is particularly important to develop a simple, practical and fast method for screening mink breeds that do not suffer from Aleutian disease.

Method used

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  • Specific primer for detecting Aleutian disease virus of minks and application of specific primer in screening healthy minks
  • Specific primer for detecting Aleutian disease virus of minks and application of specific primer in screening healthy minks
  • Specific primer for detecting Aleutian disease virus of minks and application of specific primer in screening healthy minks

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Application of Specific Primers

[0020] 1. Materials and methods

[0021] 1.1 Donor mink and reagents

[0022] 672 mink anal feces swabs were collected from many mink farms in Dalian, Changchun, Harbin, and Shandong. 2×EasyTaq PCR SuperMix was purchased from Quanshijin Biotechnology Co., Ltd. (Beijing), and DNA Marker DL2000 was purchased from Baobio Bioengineering Co., Ltd. Company (Dalian), throat swab virus DNA / RNA extraction kit was purchased from Kangwei Reagent Biotechnology Co., Ltd. (Beijing).

[0023] 1.2 Extraction of total viral DNA from mink feces

[0024] Use a dry cotton swab to collect feces from the anus of the mink, and quickly dissolve it in pure water. After freezing and thawing the swab once, extract the total amount according to the instructions of the throat swab virus DNA / RNA extraction kit (Kangwei Reagent Biotechnology Co., Ltd.). DNA.

[0025] 1.3 PCR detection of Aleutian virus in mink feces

[0026] According to the Aleutian ...

Embodiment 2

[0034] Example 2: The Consistency Experiment of the Results of Using Specific Primers to Screen Minks Infected with Aleutian Disease

[0035] In order to identify the consistency and stability of the detection results of mink infected with Aleutian virus by this PCR method, we took samples from the same mink at different time periods, collected the feces of the same mink in August, September and October, and extracted DNA Finally, specific PCR amplification was carried out, and the experiments in the 11 minks tested proved that the three PCR results were completely consistent, indicating that this method can stably detect the mink Aleutian virus, and it can imply that the Aleutian virus-infected mink is a long-term disease. Indeed, this result is consistent with literature reports.

[0036] Table 1 Consistency of application-specific PCR amplification screening in different growth stages of mink

[0037]

[0038]

Embodiment 3

[0039] Embodiment three: Application of specific primers to screen different kinds of mink

[0040] At present, the mink bred in my country mainly include the following species: American short-haired black mink, silver-blue mink, red-eyed white mink, pearl-colored mink, brown-colored mink and beige mink. Using this PCR detection method to detect Aleutian virus infection in different populations of minks in a mink farm, 100 male minks were taken from each population, and the results found that the Aleutian virus infection rate of minks in most populations reached 65% to 80%. Red-eyed ermine and American short-haired black had the highest infection rates.

[0041]

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Abstract

The invention discloses a specific primer for detecting Aleutian disease virus of minks and an application of the specific primer in screening healthy mink. The invention aims at designing a high-sensitivity specific detection primer for the Aleutian disease virus of a mink and applying the primer to the screening of minks not affected by the Aleutian disease. By rapidly and accurately detecting whether Aleutian disease virus is carried in the waste of mink, the method disclosed by the invention can be used for judging whether a host mink is suitable to be used as a breeding mink, so that complex detection by virtue of a contraimmunoelectrophoresis (CIEP) method is avoided. The screening method is stable and effective, and is nearly applicable to all varieties of minks and is high in coincidence rate with CIEP result; and on the precondition of not hurting the mink, the method can be used for rapidly detecting out positive minks carrying the virus, and the method is nearly suitable for all simple laboratories.

Description

Background technique [0001] Mink Aleutian disease (AMD) is a chronic progressive infectious disease caused by the Aleutian virus. The disease is characterized by a long incubation period, chronic course, vertical infection, abnormally increased gamma globulin in the blood, and increased plasma cells , frequent arteritis, glomerulonephritis and hepatitis, acute abortion (Best SM et al., 2005). Therefore, it is also called plasmacytosis or agammaglobulinemia. In classification, AMDV belongs to Parvoviridae, Parvovirinae, Aleutian virus, and is a single-stranded linear DNA virus. [0002] A large number of documents show that Aleutian disease has become an important infectious disease widely prevalent in various mink countries in the world (Knuuttila et al.2009); in Heilongjiang in 1973, a field in Heilongjiang used iodine reaction to check 1300 mink, and the positive rate reached 22%. Jilin Agricultural University successively inspected 12,000 mink farms in Beijing, Dalian and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/701
Inventor 易立程悦宁程世鹏陈立志仝明薇岳志刚王建科赵航林鹏
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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