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Kit used for detecting sheep pox virus and detection method thereof

A sheeppox virus and a detection method technology are applied to a kit for detecting sheeppox virus and the detection field thereof, which can solve the problems of complex operation of the kit, harm to human health, time-consuming and laborious, and achieve low cost and good stability. , the effect of high sensitivity

Active Publication Date: 2013-09-04
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, there is only an ordinary PCR detection kit used to detect sheeppox virus in the market at present. The operation of this kit is relatively complicated. Human health, please be careful when handling

Method used

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  • Kit used for detecting sheep pox virus and detection method thereof
  • Kit used for detecting sheep pox virus and detection method thereof
  • Kit used for detecting sheep pox virus and detection method thereof

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Embodiment Construction

[0030] The invention provides a detection kit for sheeppox virus, which consists of the following components:

[0031] (1) Premix Ex Taq, purchased from Bao Biological Engineering (Dalian) Co., Ltd., model RR390Q.

[0032] (2) Positive control

[0033] The positive control of this kit invention is a plasmid containing the P32 gene of sheep pox virus. The preparation method of the plasmid containing the P32 gene of sheep pox virus is as follows: extract the viral DNA from the sheep pox virus isolated from the cells, amplify to obtain the P32 gene, Cloned into pMD-18T, identified by sequencing, the plasmid with the correct inserted sequence was named pMD-P32. The concentration of the plasmid measured by Nanodrop2000 nucleic acid protein spectrophotometer was 126ng / μL, and the copy number concentration was calculated to be 3.93×10 7 copies / μL (copies / μL is copy / microliter, the same below), the positive control plasmid concentration used in the kit of the present invention is 3....

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Abstract

The invention provides a detection method used for detecting sheep pox virus, wherein real-time fluorescent quantitative PCR (polymerase chain reaction) detection is carried out on a DNA (deoxyribose nucleic acid) template of a sample by designing a pair of primers and one probe; and the invention also provides a kit used for detecting the sheep pox virus by applying the detection method. The kit used for detecting the sheep pox virus designs two specific primers aiming at P32 and one probe, real-time quantitative PCR detection is carried out on sheep pox virus Taqman, and a real-time quantitative method established aiming at the gene has high specificity and good stability; the kit used for detecting the sheep pox virus is easy to operate, a user only needs to add DNA of a sample to be inspected into a reaction tube, a detection result can be obtained according to an amplification curve, time and labour are saved, cost is lower, and time is shortened to 1-2 hours from the original 3-4 hours; and the kit used for detecting the sheep pox virus has higher sensitivity compared with common PCR and can be used for detecting the sheep pox virus with low content.

Description

technical field [0001] The invention relates to a kit for detecting goat pox virus and a detection method thereof, in particular to a kit for detecting goat pox virus by using TaqMan real-time fluorescent quantitative PCR and a detection method thereof. Background technique [0002] Goat pox (Capripox) is caused by sheep pox virus (Capripoxvirus, CPV) and can infect goats, sheep and cattle. An acute, febrile, contagious infectious disease, mainly manifested as fever, papules and herpes on the skin or mucous membranes of hairless or less hairy parts. Sheeppox is the most serious of all animal pox diseases, with a high mortality rate It can cause huge economic losses and seriously affect the development of international trade and sheep farming. Among them, sheeppox is widely prevalent in Africa, Turkey, India, and some Asian countries, and it is also sporadic in Greece. Therefore, the World Organization for Animal Health (OIE) will The disease is classified as a class A disease,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/94
Inventor 刘湘涛田宏吴锦艳陈妍尚佑军张克山尹双辉王光祥杨顺利刘永杰
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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