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Primer and probe for detecting goose circovirus

A circovirus and probe technology, applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc., to achieve good repeatability, strong specificity, and high sensitivity

Inactive Publication Date: 2011-01-12
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no fluorescent quantitative PCR method for the detection of goose circovirus at home and abroad, but there are corresponding fluorescent quantitative PCR detection methods for other types of viruses in this genus, such as porcine circovirus and pigeon circovirus

Method used

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  • Primer and probe for detecting goose circovirus
  • Primer and probe for detecting goose circovirus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 The establishment of the fluorescent quantitative PCR method of goose circovirus

[0026] 1. Materials:

[0027] Goose circovirus (GenBank accession number: GU320569)

[0028] GPV, APMV-1, and GRV viruses are preserved in the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences.

[0029] Two, steps

[0030] 1. Instruments and reagents: Mastercycler ep realplex fluorescent quantitative PCR instrument, purchased from eppendorf; PrimeScript TM RT-PCR Kit (Perfect Real Time) was purchased from Bao Biological Engineering (Dalian) Co., Ltd., PrimeScript TM RT reagent Kit, pMD18-T vector, and DL2000 Marker were purchased from Bao Bioengineering (Dalian) Co., Ltd.; gel recovery kit and plasmid mini-extraction kit were purchased from Omega Bio-Tek; GoTaq Master Green Mix was purchased from Promega; Fluorescent quantitative PCR tubes were purchased from Axygen.

[0031] 2. Specificity of Primers and Probes

[0032] The...

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Abstract

The invention provides a primer and a probe for detecting goose circovirus. The sequence of the primer is: the upstream primer: 5'-AGGTAATGGTTGCTTCTACTGTTAG-3', and the downstream primer: 5'-GTGGTATTCTACGGAATGGGTATG-3'; and the sequence of the probe is 5'-ACAAGCCGAGGACACAGCAGCACC-3'. The primer and probe are designed according to complete gene sequences of all goose circovirus, which are registered in GenBank, the fluorescent quantitative PCR method of goose circovirus is established domestically and internationally for the first time, and the method has quite high sensitivity, and the lowest detection level is 453 copies; and the method is also high in specificity and repeatability.

Description

technical field [0001] The invention relates to a primer and a probe for detecting goose circovirus. Background technique [0002] Circovirus is an animal virus with no envelope and icosahedral symmetry. Its structure is simple, and its replication depends on the vitality of host cells. Lymphocytes with fast proliferation are the main hosts of circovirus infection. Therefore, circovirus infection often leads to immune disorder and decreased immunity of the body, which in turn causes secondary infection of various conditional pathogens. It mainly invades the immune system, can cause virus-induced lymphoid tissue damage, and makes animals vulnerable to concurrent and secondary infections of other pathogens. [0003] Goose circovirus (GoCV) was first observed in 1999 by German scholar Soike et al. from diseased goose pathological tissue. The sick goose mainly showed dysplasia, weight loss, messy feathers, etc. Histopathological examination found that the lymphocytes in the bu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
Inventor 万春和黄瑜施少华傅光华程龙飞陈红梅林芳林建生
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI