African swine fever polymerase chain reaction (PCR) detection method and oligonucleotide primer pair

An African swine fever, oligonucleotide technology

Active Publication Date: 2013-09-25
QINGDAO AGRI UNIV
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Problems solved by technology

At present, African swine fever has not been introduced into my country, and there is no effective vaccine in the world to prevent and control it. In order to pr...

Method used

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  • African swine fever polymerase chain reaction (PCR) detection method and oligonucleotide primer pair
  • African swine fever polymerase chain reaction (PCR) detection method and oligonucleotide primer pair
  • African swine fever polymerase chain reaction (PCR) detection method and oligonucleotide primer pair

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Experimental program
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Effect test

Embodiment

[0017] 1.1 ASFV P72 plasmid

[0018] Using conventional PCR technology, according to the nucleotide sequence of the African swine fever virus P72 gene published by GeneBank, the full-length ASFV P72 gene was amplified and inserted into the pcDNA3.1 vector.

[0019] 1.2 Source and treatment of tissue disease materials

[0020] Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), and porcine circovirus type 2 (PCV2) were purchased from Harbin Veken Biotechnology Development Company.

[0021] Clinical samples were collected from sows with reproductive failure or piglets with symptoms of respiratory impairment and aborted fetuses from 14 farms.

[0022] Aseptic treatment of clinical tissue disease materials: cut 2.0g~4.0g of tissue disease materials, add a small amount of sterilized PBS (containing penicillin and streptomycin), cut them into pieces, grind them into paste with a sterile grinder, and press The volume ratio of 1:4 was dil...

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Abstract

The invention belongs to the technical field of biological detection and relates to an African swine fever polymerase chain reaction (PCR) detection method and an oligonucleotide primer pair. The method comprises the following steps: designing a pair of specific primers by referring to a complete genome sequence of 23 ASFVP72 protein strains recorded in GenBank, exploring the optimal reaction system and reaction conditions according to the designed primers, performing sensitivity test, specificity test and reproducibility test on the established PCR reaction method, and performing PCR method detection on clinical samples of sows suffered from reproductive disturbance or piglets with respiratory disturbance symptoms and aborted fetuses from 14 pig farms. The test results prove that the established PCR method can sensitively and rapidly detect the African swine fever virus. The PCR detection method established in the research has high applicability and can be used for detecting the African swine fever disease.

Description

technical field [0001] The invention relates to a PCR method and an oligonucleotide primer pair for detecting African swine fever, belonging to the technical field of biological detection. Background technique [0002] African swine fever (ASF) is an acute, febrile, highly contagious disease of pigs caused by African swine fever virus. It is extremely harmful to the pig industry and is an infectious disease that is strictly guarded against in various countries. It is listed as a first-class animal disease in my country. The clinical symptoms and pathological changes are high fever, skin hemorrhage, respiratory disorder, neurological symptoms, miscarriage, edema and visceral hemorrhage, all of which are similar to acute swine fever, and it is easy to misdiagnose during diagnosis. 100%. At present, African swine fever has not been introduced into my country, and there is no effective vaccine in the world to prevent and control it. In order to prevent the disease from being i...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 单虎张传美秦晓冰王述柏黄娟杨瑞梅陈琳琳
Owner QINGDAO AGRI UNIV
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