Goat pox virus/contagious pustular dermatitis virus multiplex PCR (polymerase chain reaction) detection primers

A technology of oral ulcer virus and goat pox virus, which is applied in the direction of microorganisms, recombinant DNA technology, and methods based on microorganisms, can solve the problem of low sensitivity and achieve the effect of shortening the detection time

Inactive Publication Date: 2016-09-07
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this method can detect two viruses at the same time, the sens

Method used

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  • Goat pox virus/contagious pustular dermatitis virus multiplex PCR (polymerase chain reaction) detection primers
  • Goat pox virus/contagious pustular dermatitis virus multiplex PCR (polymerase chain reaction) detection primers
  • Goat pox virus/contagious pustular dermatitis virus multiplex PCR (polymerase chain reaction) detection primers

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Experimental program
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Embodiment Construction

[0015] Concrete technical scheme of the present invention is as follows:

[0016] (1) Design of primers

[0017] According to the purpose of the experiment, after homology comparison, the conserved gene regions of goat pox virus and aphthus virus were selected, and specific primers were designed. The primer sequences are as follows:

[0018]

[0019] Note: Merged base code R=A / G Y=C / T

[0020] (2) Extraction of viral DNA

[0021] Take about 5mg of animal tissue (collected lesion tissue), put it into a 2mL tube, add an appropriate volume of four-antibody PBS (containing 10000U / mL penicillin, 10mg / mL streptomycin, 250μg / mL gentamicin, 250μg / mL kanamycin), grind with a tissue homogenizer, centrifuge to take 500 μL of the supernatant, add 92 μL of 10% SDS and 8 μL of proteinase K, mix thoroughly, 58 ° C metal bath for 30 min, and then extract with an equal volume of phenol chloroform for 2 Once, pipette 400 μL supernatant into a new EP tube, add 40 μL sodium acetate soluti...

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Abstract

The invention relates to goat pox virus/contagious pustular dermatitis virus multiplex PCR (polymerase chain reaction) detection primers of which the sequences are disclosed as SEQ ID No.1-4. The 2 pairs of primers are utilized to carry out PCR amplification on the corresponding target genes so as to establish a multiplex PCR detection method; and the primers can be used for quickly identifying goat pox virus and contagious pustular dermatitis virus in one reaction system, and can simultaneously determine whether two-virus mixed infection occurs, thereby having important application value in the aspects of clinical sample quick screening, disease quick diagnosis and the like.

Description

technical field [0001] The invention relates to a primer for detecting goat pox virus and oropharynx virus by multiplex PCR. Background technique [0002] Goat pox (Goat pox, GP) is an acute contagious disease caused by goat pox virus (GTPV), and papular-pustular pox occurs on the skin. It can infect goats of all breeds, genders and ages. It has been reported that sheep can also be infected, often causing abortion of pregnant sheep and huge economic losses. It is one of the animal diseases that must be declared under the OIE regulations. Oral aphthous is an acute contact and epithelial infectious disease of sheep and goats caused by Orf virus (ORFV). It is characterized by the formation of papules, blisters, pustules, ulcers and thick warty scabs on the skin and mucous membranes of the mouth, lips, tongue, nose, and breasts. Although it can recover on its own, secondary infection and slow growth seriously affect the development of the breeding industry. [0003] In recent...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2537/143
Inventor 陈素娟彭大新秦涛朱相儒易成功
Owner YANGZHOU UNIV
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