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Kit for testing Marek's disease virus of chicken by using MEQ gene and detection method thereof

A Marek's and genetic detection technology, applied in the detection field of chicken Marek's virus, can solve the problem that the detection method cannot meet the needs of fast, sensitive and accurate detection, and achieve low cost, high sensitivity and good specificity

Inactive Publication Date: 2013-10-23
SICHUAN AGRI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0007] The technical problem to be solved in the present invention is to overcome the existing detection method that cannot adapt to the fast, sensitive and accurate detection requirements, thereby providing a kind of reagent that utilizes the MEQ gene to detect chicken Marek's virus that can detect the virus quickly, sensitively and accurately Box and method for detecting chicken Marek's virus using the kit

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  • Kit for testing Marek's disease virus of chicken by using MEQ gene and detection method thereof
  • Kit for testing Marek's disease virus of chicken by using MEQ gene and detection method thereof
  • Kit for testing Marek's disease virus of chicken by using MEQ gene and detection method thereof

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Embodiment Construction

[0031] The present invention is specifically described below in conjunction with specific samples:

[0032] (1) The acquisition of samples and the establishment of control groups;

[0033] Positive individuals of AA broiler chickens, Roman layer hens, and Erlangshan mountain chickens that were found to be infected with Marek's virus during the breeding process were collected as samples, and uninfected individuals were also from the poultry farm of Sichuan Agricultural University as the control group.

[0034] (2) Tissues such as liver, spleen, kidney, bursa and thymus were collected from each chicken in the infection group and the control group. After the samples were collected, the EP tubes containing the tissue samples were stored in liquid nitrogen, and then transferred to a -80°C refrigerator.

[0035] (3) Extract the viral DNA from the collected tissue according to the operating instructions of the Baobio Virus DNA Extraction Kit;

[0036] (4) Design primers according t...

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Abstract

The invention relates to a test method of Marek 's disease virus of chicken, specifically a kit for testing Marek's disease virus of chicken by using MEQ gene, and also includes a test method of the kit. The kit includes a virus genomic DNA extraction reagent, Master Mix, an SYBR Green I premixed liquid reagent, negative control, and positive control. The kit also includes two specific primers, sequences of which are MEQ-F:5'-ACGCAGGGAGCAGACGTACTAT-3' and MEQ-R:5'-CCATAGGGCAAACTGGCTCAT-3'. The kit is high in sensitivity, the plasmid with a concentration of 1.17*10<2> copies / [mu]L which is obtained by diluting standard plasmid to 10-fold still shows a good fluorescence amplification curve, good specificity and stability are shown through the design of primers and optimization for PCR reaction system, and the copy number of a sample to be tested can be calculated too. The kid and the method have the advantages of quick and easy operation, low cost, and stable and accurate test results.

Description

technical field [0001] The invention relates to a detection method of chicken Marek's virus, specifically a kit for detecting chicken Marek's virus by using MEQ gene, and the invention also includes the detection method of the kit. Background technique [0002] Marek's disease [0003] Marek's disease (Marek's Disease, MD) is a highly contagious and highly harmful tumor disease, which is the most common chicken malignant tumor disease (cancer) caused by the Marek's disease herpes virus in the B subgroup of the herpesviridae. This disease is quite harmful, for example, it is highly contagious. It occurs in the United States, China and many other countries and causes great economic losses. %, and can cause death. In the prevention and treatment of this disease, there is no specific drug treatment; in the prevention, vaccination and genetic resistance can be adopted. Marek virus is divided into three serotypes, serotype 1 (MDV-1) is mainly a tumorigenic strain and an attenua...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 程洋冯泽清刘益平朱庆金洁
Owner SICHUAN AGRI UNIV