General RT-PCR detection primer for muscovy duck reoviruses and detection method

A technology of RT-PCR and reovirus, which is applied in the field of Muscovy duck virus detection, can solve the problems that the RT-PCR detection method of Muscovy duck reovirus cannot be established, and achieve high sensitivity, good repeatability and specificity strong effect

Active Publication Date: 2014-08-06
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, the RT-PCR method established above is designed based on differential diagnosis, and can only amplify the bands of the respective viruses, and detect MDRV or NDRV alone, and cannot establish a general RT-PCR detection method for Muscovy duck reovirus

Method used

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  • General RT-PCR detection primer for muscovy duck reoviruses and detection method
  • General RT-PCR detection primer for muscovy duck reoviruses and detection method
  • General RT-PCR detection primer for muscovy duck reoviruses and detection method

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Embodiment 1

[0031] 1. Test material

[0032] 1.1 virus

[0033] MDRV isolates ARV-00m and ARV-05m were isolated from young muscovy ducks suffering from "white spot disease" in Yuhang District, Hangzhou City in October 2000 and Lanxi City in June 2005; NDRV isolate ARV-03g It was isolated from goslings with "hemorrhagic necrotizing hepatitis" in February 2003. ARV-HNF was isolated from Muscovy ducks in Haining City with "hemorrhagic necrotizing hepatitis" in August 2010. ARV-HNF -09w was isolated from a Hangzhou wild duck that suffered from "hemorrhagic necrotizing hepatitis" in 2009.

[0034] In addition, duck Tambusu virus (DTMUV) strain YY5, duck plague virus (DEV) vaccine strain, goose-derived Newcastle disease virus (NDV) DQ strain, Muscovy duck parvovirus (MDPV) and duck hepatitis A virus ( DHAV), Egg Drop Syndrome Virus (EDSV) and H9N2 Subtype Avian Influenza Virus (AIV) were all provided and preserved by the Institute of Animal Husbandry, Zhejiang Academy of Agricultural Sciences...

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Abstract

The invention discloses a general RT-PCR detection primer for muscovy duck reoviruses and a detection method, and belongs to the technical field of muscovy duck reovirus detection. The general RT-PCR detection primer comprises an upstream primer, namely 5'-CACAGTGCTACCATC-3', and a downstream primer, namely 5'-CTGCTGATCATAATT-3'. The detection method comprises the following steps: RT-PCR amplification is carried out by using the general RT-PCR detection primer for muscovy duck reoviruses and samples RNA to be detected, and if the amplification product comprises 436bp fragments, the samples to be detected comprise the muscovy duck reoviruses. After comparison according to the sigma A gene sequence of the muscovy duck reovirus, the general primer is optimally designed, and the one-step process RT-PCR is optimized further. The detection method not only can detect the MDRV muscovy duck reovirus and the NDRV muscovy duck reovirus at the same time, but also has the characteristics of being good in repeatability, strong in specificity, and high in sensibility.

Description

technical field [0001] The invention relates to the technical field of muscovy duck virus detection, in particular to a universal RT-PCR detection primer and a detection method for muscovy duck reovirus. Background technique [0002] Muscovy duck reovirus disease is an acute infectious disease of muscovy ducks, muscovy ducks and geese caused by Muscovy duck reovirus (MDRV). The main clinical lesions are soft feet, diarrhea and growth Dysplasia, with a large number of gray-white necrotic spots on the surface of liver and spleen as the main lesion manifestation, commonly known as "white spot disease" or "flower liver disease" of Muscovy duck liver, this disease mainly occurs in Muscovy ducks aged 10-45 days, and the incidence is usually 30% to 90%, and the case fatality rate is generally 60% to 80%. In severe cases, it can lead to the death of the whole group, causing huge economic losses in the Muscovy duck breeding industry. [0003] In 1972, French scholars isolated and id...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/686C12Q1/70C12Q2531/113
Inventor 叶伟成余斌张存云涛陈柳倪征华炯钢
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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