Primer and probe combination for detecting Newcastle disease virus, kit and detection method

A technology of Newcastle disease virus and kit, which is applied in the field of detection of primers and probe combinations of Newcastle disease virus, and can solve the problems of long detection time and high technical requirements

Pending Publication Date: 2020-05-08
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problems that existing PCR and RFQ-PCR require longer detection time and higher technical requiremen

Method used

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  • Primer and probe combination for detecting Newcastle disease virus, kit and detection method
  • Primer and probe combination for detecting Newcastle disease virus, kit and detection method
  • Primer and probe combination for detecting Newcastle disease virus, kit and detection method

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Effect test

Embodiment 1

[0031] The embodiment of the present invention provides a combination of primers and probes for detecting Newcastle disease virus, the sequences are respectively:

[0032] Upstream primer: 5′-ACGTCTTTATTACTCCTGAGCTTGTCATTGTGA-3′;

[0033] Downstream primer: 5′-TGATTACCTAAGTCCTTTGCCAGAGCATCTACT-3′;

[0034] Probe: 5'-AAGTTCACATGCCTCACCCAGGAACTTGTA-THF-TGATGTATGCGGATA-3'.

Embodiment 2

[0036] Embodiments of the present invention provide a kit for detecting Newcastle disease virus, specifically comprising:

[0037] (1) Isothermal amplification primers and probes: the sequences are respectively:

[0038] Upstream primer: 5′-ACGTCTTTATTACTCCTGAGCTTGTCATTGTGA-3′;

[0039] Downstream primer: 5′-TGATTACCTAAGTCCTTTGCCAGAGCATCTACT-3′;

[0040] Probe: 5'-AAGTTCACATGCCTCACCCAGGAACTTGTA-THF-TGATGTATGCGGATA-3'.

[0041] (2) Fluorescence basic buffer, magnesium acetate (both are RT-RAA nucleic acid amplification reagents of Jiangsu Qitian Gene Biotechnology Co., Ltd.).

Embodiment 3

[0043] The embodiment of the present invention provides a method for detecting Newcastle disease virus, which is operated with the kit in embodiment 2.

[0044] The samples to be tested are:

[0045] Both NDV and avian influenza virus (Avian Influenza Virus, AIV) were preserved in the laboratory of Hebei Agricultural University, chicken infectious laryngotracheitis virus (Infectious Laryngotracheitis Virus, ILTV) AV195 and chicken infectious bronchitis virus (Infectious Bronchitis Virus, IBV) AV1511 was purchased from China Veterinary Drug Control Institute. Negative control (N) used pure water instead of template. The steps are as follows:

[0046] (1) Recombinase-mediated isothermal nucleic acid amplification reaction

[0047] Use the DNA / RNA extraction kit (purchased from Tiangen Biochemical Technology Co., Ltd.) to extract the RNA of NDV, AIV, IBV and the DNA of ILTV respectively, and use the cDNA obtained by reverse transcription as a template, and recombine according ...

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Abstract

The invention relates to the technical field of virus detection, specifically to a primer and probe combination for detecting Newcastle disease virus, a kit and a detection method. A to-be-detected object containing a Newcastle disease virus RNA fragment can be massively amplified under an isothermal condition through a reverse transcription recombinase mediated isothermal nucleic acid amplification technology by utilizing the primer and probe combination; and the primer and probe combination have the advantages of simple operation, short required time, good specificity and high sensitivity.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a combination of primers and probes for detecting Newcastle disease virus, a kit and a detection method. Background technique [0002] Newcastle Disease (ND), also known as pseudo fowl plague, is an acute and highly contagious disease of poultry caused by Newcastle disease virus (NDV) infection with the digestive tract and respiratory tract as the main transmission route. All can fall ill in four seasons, very harmful to the poultry breeding industry in the world. NDV mainly infects poultry, chickens are the most common, and chicks are the most susceptible. The clinical manifestations of sick chickens are increased body temperature, shortness of breath, coughing, and discharge of yellow-green or even gray-white watery feces. The egg production rate of laying hens decreases, and the proportion of soft-shell eggs increases. Therefore bring serious economic loss to aquacul...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2531/119C12Q2521/507C12Q2565/625C12Q2521/107C12Q2563/107
Inventor 王春光王文静张铁翟向和李希明曹立辉白云徐瑞涛
Owner HEBEI AGRICULTURAL UNIV.
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