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PCR method for quick identifying newcastle disease virus, and indentifying strong or weak viral strain

A technology for Newcastle disease virus and attenuated strains, applied in the field of virus identification, can solve the problems of long reporting time and heavy workload, and achieve the effect of rapid virulence identification test method

Inactive Publication Date: 2008-05-21
YANGZHOU UNIV
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  • Claims
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Problems solved by technology

These methods take a long time to report results and require a lot of work

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  • PCR method for quick identifying newcastle disease virus, and indentifying strong or weak viral strain
  • PCR method for quick identifying newcastle disease virus, and indentifying strong or weak viral strain
  • PCR method for quick identifying newcastle disease virus, and indentifying strong or weak viral strain

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Embodiment Construction

[0016] 1. Primer design:

[0017] Four specific primers (Primer I, Primer II, Primer III, Primer IV) were designed according to the conserved sequence of the Newcastle disease virus fusion protein (F) gene published in the GenBank database. synthesis.

[0018] In addition, it has been reported in GenBank: Newcastle disease virus fragment length: 570bp; Newcastle disease virus strong strain fragment length: 200bp; Newcastle disease virus attenuated strain fragment length: 395bp; indicator band (Escherichia coli) fragment length: 750bp .

[0019] The size of the expected amplified target fragment is divided into Newcastle disease virus: 570bp; Newcastle disease virus strong strain: 200bp; Newcastle disease virus attenuated strain: 395bp; indicator band (E. coli): 750bp.

[0020] Newcastle disease virus:

[0021] Primer I: 5'-[TA(C / T)ACCTC(A / G)TC(T / C)CAGAC(A / T)GG]-3'

[0022] Primer II: 5'-[CCACCAGC(T / C)A(G / A)ATT(G / A)TAAAG]-3'

[0023] Virulent strains of Newcastle disease v...

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Abstract

The PCR method for identifying the Newcastle disease virus fast and differentiating whether the strain is a high virulent one or a low virulent one relates to a method for identifying virus, in particular to a method for identifying the high or low virulent strain of the Newcastle disease virus. Trizol kit is used for extracting the RNA of the sample gene for inverse transcription and composition of cDNA; at the same time, the method of thermal cracking is adopted for extracting the DNA template of colon bacillus and then a plurality of special genes is magnified in the PCR pattern for one time on the cDNA of the sample gene and the DNA template of the colon bacillus and finally the gene is identified through an electrophoresis process. The invention forms a large number of gene copy segments which is convenient for identifying whether the virus is the Newcastle disease one for one time and whether the strain is a high virulent one or a low virulent one. The invention is faster and more accurate than the classical method of virus segregation, identification and virulence identification and experiment.

Description

Technical field: [0001] The invention discloses a detection method for identifying viruses, in particular for identifying strong and weak strains of Newcastle disease virus. Background technique: [0002] Newcastle disease (ND) is an acute and highly contagious poultry disease caused by Newcastle disease virus (NDV), which is a serious hazard to the poultry industry. The World Organization for Animal Health (OIE) lists it as a statutory notifiable infectious disease, and my country also defines it as a first-class animal infectious disease. [0003] With the development of highly intensive poultry farming, the control of ND has attracted much attention. The on-site diagnosis of ND mainly relies on the comprehensive judgment of history, clinical symptoms, pathological changes and epidemiological investigation, which has brought overload work to the majority of veterinary staff, consuming time, effort and money. Rapid and accurate screening and diagnosis methods are urgently...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 焦新安潘志明耿士忠黄金林顾志强孙林张磊殷月兰
Owner YANGZHOU UNIV