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Real-time fluorescence quantification PCR detection primer for quickly identifying genotypes of duck circovirus and detection method thereof

A technology of real-time fluorescence quantification and duck circovirus, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, to achieve fast accuracy, fast and accurate detection, and simple identification methods

Pending Publication Date: 2016-11-02
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

However, there is no report on the real-time fluorescent quantitative PCR (real-timequantitative PCR, qPCR) method for the differential diagnosis of genotype 1 and genotype 2 duck circovirus, especially only one set of primers is needed for genotype 1 and genotype 2. The research report on the kit and method for the identification of genotype 2 duck circovirus, the present invention can fill in the gaps in related fields

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  • Real-time fluorescence quantification PCR detection primer for quickly identifying genotypes of duck circovirus and detection method thereof
  • Real-time fluorescence quantification PCR detection primer for quickly identifying genotypes of duck circovirus and detection method thereof
  • Real-time fluorescence quantification PCR detection primer for quickly identifying genotypes of duck circovirus and detection method thereof

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Embodiment Construction

[0030] The content of the present invention will be described in detail below in conjunction with the drawings and embodiments of the specification:

[0031] A primer for rapid identification of duck circovirus genotype by real-time fluorescent quantitative PCR detection, the sequence of the primer is as follows:

[0032] Upstream primer P1: 5'-CAGATCCCCGGGCACGAGA-3';

[0033] Downstream primer P2: 5'-CCTCACCTTCAGGGATTC-3'.

[0034] A real-time fluorescent quantitative PCR detection method for quickly identifying duck circovirus genotypes, using the upstream primer P1: 5′-CAGATCCCCGGGCACGAGA-3′ and the downstream primer P2: 5′-CCTCACCTTCAGGGATTC-3′ to establish a real-time fluorescent quantitative PCR method based on SYBRGreenⅠ The difference in the GC content of the nucleotides in the region of the specific gene fragments of genotype 1 duck circovirus and genotype 2 duck circovirus amplified by the primers leads to a difference in melting curve temperature, and the difference in melt...

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Abstract

The invention relates to a real-time fluorescence quantification PCR detection primer for quickly identifying genotypes of duck circovirus and a detection method thereof. The sequences of the primer are described as follows: an upstream primer P1: 5'-CAGATCCCCGGGCACGAGA-3'; a downstream primer P2: 5'-CCTCACCTTCAGGGATTC-3'. The detection method includes the steps of: establishing the real-time fluorescence quantification PCR method based on SYBR Green I with the primer, wherein difference of temperature of a melting curve exists due to the difference of GC content of a nucleotide in a specific gene fragment region of gene 1-type duck circovirus and gene 2-type duck circovirus which are amplified by the primer, so that infection of the gene 1-type duck circovirus and the gene 2-type duck circovirus can be directly carried out according to the difference of Tm value in the melting curve. The method can distinguish the infection caused by the duck circovirus in different genotypes just with one group of primer, thereby providing technical fundament for healthy breeding in duck breeding industry.

Description

Technical field [0001] The invention relates to the technical field of rapid diagnosis of animal infectious diseases, in particular to a real-time fluorescent quantitative PCR detection primer for quickly identifying duck circovirus genotypes and a detection method thereof. Background technique [0002] Duck circovirus (DuCV) is a new virus in duck flocks in recent years, and all kinds of ducks can be infected. It was first reported by Hattermann et al. in 2003 and the earliest report by Fu Guanghua et al. (DuCV-MH25 strain, GenBank number EF451157). The clinical symptoms manifest as skin and lymphatic system damage, which makes the animal body vulnerable to concurrent and secondary infections by other pathogens; there are also symptoms manifested as abnormal development and feather disorder; and can infect the immune system of ducks, causing immune suppression of sick ducks, and feeding The duck industry caused significant economic losses. [0003] Duck circovirus is a non-envel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2531/113C12Q2563/107
Inventor 陈翠腾陈珍朱春华刘斌琼蔡国漳
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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