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Method for detecting newcastle disease virus nucleic acid by RT-PCR

An RT-PCR and Newcastle disease virus technology, applied in the field of Newcastle disease virus nucleic acid RT-PCR detection, can solve problems such as easy missed detection, and achieve the effect of reducing detection time, accurate detection, and solving easy missed detection.

Inactive Publication Date: 2015-01-14
POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
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Problems solved by technology

It is relatively time-saving to directly take feces to detect viral nucleic acids, but due to the small amount of sample poisoning, it is easy to miss detection

Method used

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  • Method for detecting newcastle disease virus nucleic acid by RT-PCR
  • Method for detecting newcastle disease virus nucleic acid by RT-PCR
  • Method for detecting newcastle disease virus nucleic acid by RT-PCR

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Embodiment Construction

[0021] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention, but the present invention is not limited thereby.

[0022] A method for Newcastle disease virus nucleic acid RT-PCR detection, comprising the following steps:

[0023] (1) Virus proliferation: Take the animal feces to be tested, add the animal feces and normal saline with a mass ratio of 1:3 into a centrifuge tube and shake fully to form a suspension, and add penicillin and streptomycin to each ml of the suspension. 1000 units, centrifuge at 8000g after 4-6 hours of reaction, take sterile supernatant, inoculate 9-11-day-old chicken embryos with the supernatant through the allantoic cavity, discard dead chicken embryos within 24 hours, and collect 24-96-hour chickens Embryo allantoic fluid, reserve at -20°C;

[0024] (2) Design primers: with reference to the F gene sequ...

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Abstract

The invention relates to a virus detection method, and particularly relates to a method for detecting newcastle disease virus nucleic acid by RT-PCR, which comprises virus multiplication, primer design, nucleic acid extraction, product identification, and sequence identification; the beneficial effects of the invention are that: according to the method, virus multiplication is performed firstly by chick embryo inoculation, and then PCR detection is performed; an R degenerate primer is added into an F-F primer, which realizes the detection of week viruses and strong viruses during the detection of the newcastle disease virus nucleic acid; the detection is accurate, reduces the time for respectively detecting week viruses and strong viruses, and solves the problem of easy detection missing in traditional methods.

Description

technical field [0001] The invention relates to a virus detection method, more specifically a method for RT-PCR detection of Newcastle disease virus nucleic acid. Background technique [0002] Newcastle disease ((Newcastle disease, ND) is caused by Newcastle disease virus (Newcastle disease virus, NDV) a kind of acute, severe contagious disease of poultry, brings great economic loss to poultry industry and rare poultry industry. In addition, from many NDV has also been isolated from species of wild birds such as sparrows, pigeons, crows, etc. Fusion protein F is necessary for NDV to infect cells. The fusion of the virus envelope with the target cell allows the virus to enter the cell for replication. The fusion process relies on the F protein A large number of studies have shown that the amino acid motif of the F protein F1 / F2 polypeptide cleavage site of NDV is closely related to the virulence of NDV, and its pathogenic mechanism has been confirmed by reverse genetic manipu...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 袁小远王友令张玉霞徐绍建
Owner POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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