Primers and probe for dual real-time fluorescent quantitative PCR detection of duck circovirus type 1 and duck circovirus type 2

A real-time fluorescence quantitative, duck circovirus technology, applied in the field of animal virology, can solve the problems of high sensitivity, unobserved feather mess and weight loss, etc., to achieve the effects of high sensitivity, rapid detection, and simplified operation procedures

Pending Publication Date: 2020-10-09
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

In 2018, Korean scholars studied the pathogenicity of D11-JW-008 strain (belonging to DuCV-1 branch) to Peking duck (24d) and found that apoptosis could be detected in the bursa of Peking duck after infection, but the experimental group and Similar to the control group, typical clinical symptoms of DuCV infection (such as emaciation, disheveled feathers, and weight loss, etc.) were not observed
At present, there are no research reports on real-time fluorescent quantitative primer sets and probe sets that can specifically diagnose DuCV-1 and DuCV-2 infections. This method can not only be used for epidemiological detection (reaching the effect of conventional PCR detection, But the sensitivity is higher), and it can accurately quantify the specific subtype infection of DuCV-1 and DuCV-2, and at the same time accurately distinguish different infection types

Method used

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  • Primers and probe for dual real-time fluorescent quantitative PCR detection of duck circovirus type 1 and duck circovirus type 2
  • Primers and probe for dual real-time fluorescent quantitative PCR detection of duck circovirus type 1 and duck circovirus type 2
  • Primers and probe for dual real-time fluorescent quantitative PCR detection of duck circovirus type 1 and duck circovirus type 2

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Experimental program
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Effect test

Embodiment 1

[0041] 1. Related test pathogens

[0042] 1.1 Experimental strains

[0043] Duck circovirus genotype 1 (DuCV-1, strain JX209) and duck circovirus genotype 2 (DuCV-2, strain FJ1815) were isolated, identified and preserved by the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences.

[0044] 1.2 Experimental control strains and bacterial strains

[0045] Common nucleic acid types in ducks are DNA pathogens, such as goose parvovirus (GPV), muscovy duck parvovirus (MDPV), duck adenovirus type A (DAdV-A), duck plague virus (DEV), duck Escherichia coli (E coli), Richteria anatipestifer (R.A.), and Pasteurella multocida (P.M.) from ducks were all identified and preserved by the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences.

[0046] 2. Primer design for TaqMan real-time fluorescent quantitative PCR detection method

[0047] 2.1 Design of primers and probes

[0048] According to the results ...

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Abstract

The invention relates to a primer and a probe for dual real-time fluorescent quantitative PCR detection of duck circovirus type 1 and duck circovirus type 2. The sequences of the primer and the probeare as shown in SEQ. ID. No.1 to SEQ.ID. No.6. A dual real-time fluorescent quantitative PCR detection method established by utilizing the primers and the probe is high in sensitivity, good in stability, strong in specificity and good in repeatability, can be used for duck circovirus typing detection, and lays a foundation for subsequent scientific research on genotype pathogenesis of two duck circoviruses and development of molecular epidemiology.

Description

technical field [0001] The invention belongs to the field of animal virology, and in particular relates to a primer and a probe for dual real-time fluorescent quantitative PCR detection of duck circovirus type 1 and duck circovirus type 2. Background technique [0002] Real-time fluorescent quantitative PCR method (Real-time PCR) is a method of detecting the total amount of products after each cycle of polymerase chain reaction (PCR) with a fluorescent chemical substance in a DNA amplification reaction. During the PCR amplification process, real-time fluorescent quantitative PCR can detect the progress of PCR in real time through fluorescent signals. Due to the exponential phase of PCR amplification, there is a linear relationship between the Ct value of a template and the initial copy number of the template. The fluorescent probe method uses sequence-specific fluorescently labeled probes to detect products. The appearance of the probe method greatly improves the specificit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/701C12Q1/6851C12Q2600/16C12Q2531/113C12Q2537/143C12Q2561/101C12Q2563/107
Inventor 陈翠腾万春和傅光华黄瑜程龙飞施少华陈红梅傅秋玲刘荣昌陈珍朱春华
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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