Preparation and use of giant panda Ascaris schroederi antigen

A technology for Ascaris ximani and giant panda, which is applied in the field of preparation of Ascaris ximani and can solve the problems of less research on immunodiagnosis technology of Ascaris lumbricoides, etc.

Inactive Publication Date: 2009-01-21
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Compared with other insect species, there are less studies on immunodiagnostic techniques for Ascaris

Method used

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  • Preparation and use of giant panda Ascaris schroederi antigen
  • Preparation and use of giant panda Ascaris schroederi antigen
  • Preparation and use of giant panda Ascaris schroederi antigen

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Embodiment Construction

[0030] Embodiments are given and further descriptions are given in conjunction with the accompanying drawings. Firstly, collect roundworm eggs by giant panda feces precipitation method, put them in a culture dish containing 2% formalin in normal saline, cover them, and culture them in an incubator at 25-28°C, and a large amount of infectivity can be obtained in 3-4 weeks. Eggs, artificially infect white rabbits with giant panda roundworm infective eggs, and each rabbit was given 3,500 eggs, repeated every other week. Before each immunization, blood was collected from the ear vein of the experimental animal, and after four times of immunization, the titer was determined by agar expansion method. When the titer of the serum reached 1:8, blood was collected from the heart, and the separated serum was collected, divided into small tubes, and stored at -20°C for future use. The rabbit anti-Panda Ascaris positive serum was obtained for the next test. Take the collected giant panda ...

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Abstract

The invention discloses preparation and application of a giant panda Ascaris schroederi Mcintosh antigen, which belong to the giant panda Ascaris schroederi Mcintosh prevention, treatment and detection field. The method comprises the following steps: a Ascaris schroederi Mcintosh antigen gene primer is designed; total RNA of ascarids undergoes reverse transcription by the RT-PCR method to synthesize cDNA, and then the cDNA is taken as a template for PCR amplification of a target product; the purified target product is connected with a pMD18-T vector and then converted into DH5 alpha competent bacteria; positive recombinant clone is selected through flat screening and culture of the bacteria, and the antigen genes have a Bs-Ag1gene, a Bs-Ag2 gene and a Bs-Ag3 gene after culture and sequencing; and then recombinant plasmids which are accurately sequenced are converted into Escherichia coli BL21 competent cells for mass expression of proteins after construction of the recombinant plasmids, induction expression and purification of the recombinant proteins. After detection and immunologic tests of the product and ELISA detection and analysis of a test animal antibody IgG, the Bs-Ag1gene, the Bs-Ag2 gene and the Bs-Ag3 gene of the giant panda Ascaris schroederi Mcintosh antigen can be taken as candidate genes for preparing genetic engineering vaccines through the sascarids; and the recombinant proteins Bs-Ag1, Bs-Ag2 and Bs-Ag3 have good reactionogenicity and can be used for detecting infection of giant panda Ascaris by the ELISA method.

Description

technical field [0001] The invention belongs to the field of prevention and detection of roundworms, and in particular relates to the preparation and application of the giant panda ascariasis. Background technique [0002] The pathogen of giant panda ascariasis is Ascaris schroederi. Roundworms usually parasitize the small intestine. However, they have also been found in the mouth, larynx, trachea, and stomach. They can also drill into the ducts that communicate with the intestinal tract, such as the pancreatic duct and bile duct, causing obstruction and inflammation, and even death. Scholar Feng Wenhe The dead bodies of 33 wild giant pandas were dissected, and the cause of death was analyzed: 100% of wild giant pandas were infected with roundworms, and the direct and indirect mortality rate of giant pandas infected with roundworms was 66.67%, which seriously endangered the survival of giant panda species. Due to the different symptoms of ascariasis, the harm of roundworms ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/70C07K14/435A61K48/00A61P33/10G01N33/53
Inventor 杨光友何光志汪涛彭雪蓉
Owner SICHUAN AGRI UNIV
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