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Method of synchronous distinguishing newcastle disease virus and vaccine virus and identifying virulence and genotype

A newcastle disease virus and vaccine virus technology, applied in the field of automated analysis software, can solve the problems of small number of verification samples, insufficient theoretical foundation, and difficult application

Inactive Publication Date: 2007-03-14
HUZHOU TEACHERS COLLEGE
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, its theoretical basis is based on the difference of a small number of nucleotides in the primer region between a small number of virulent and attenuated nucleic acid sequences, which basically does not involve the molecular basis of Newcastle disease virus pathogenicity. The number of samples is relatively small, and the actual promotion and application are more difficult

Method used

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  • Method of synchronous distinguishing newcastle disease virus and vaccine virus and identifying virulence and genotype
  • Method of synchronous distinguishing newcastle disease virus and vaccine virus and identifying virulence and genotype
  • Method of synchronous distinguishing newcastle disease virus and vaccine virus and identifying virulence and genotype

Examples

Experimental program
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Embodiment 1

[0116] Embodiment 1-isolate the detection of Newcastle disease strain

[0117] 1. Virus isolation and identification.

[0118] According to the method of "Diagnostic Test and Vaccine Standard Manual" (OIE, 1996), 51 strains of Newcastle disease virus were isolated from geese, pigeons, chickens and ostriches.

[0119] 2. Extraction of viral nucleic acid RNA

[0120] (1) Take 100 μl of allantoic fluid to be tested.

[0121] (2) Add 900 μl TRIzol reagent (Gibco-BRL company), mix well, and keep at room temperature for 5 minutes;

[0122] (3) Add 0.2ml or appropriate amount of chloroform, vortex and mix well, then let stand at room temperature for 3 minutes;

[0123] (4) 12000g, 4°C, centrifuge for 15 minutes;

[0124] (5) Carefully draw the supernatant into a new 1.5ml centrifuge tube;

[0125] (6) Add 0.5ml or an appropriate amount of isopropanol, vortex and mix well, then place at room temperature for 10 minutes;

[0126] (7) 12000g, 4°C, centrifuge for 10 minutes;

[012...

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Abstract

The present invention discloses molecular biological method and automatic analysis software for distinguishing wild virus strain and vaccine virus strain of new castle disease virus (NDV), and identifying the toxicity and genotype simultaneously. The NDV is RT-PCR amplified with one pair of specific degenerate primers to obtain one multipurpose NDV segment for direct sequencing of the PCR product. One automatic analysis software is composed for comparing the homology of the sequence to the standard vaccine sequence so as to distinguish wild virus strain and vaccine virus strain, analyze the toxicity by means of the F gene cracking site and judge the genotype through developing tree analysis. The present invention has short detecting period, reliable detection result and capacity of obtaining several detection results simultaneously, and is suitable for use in fowl product inspector and quarantiner.

Description

technical field [0001] The invention relates to a molecular biology method capable of synchronously distinguishing wild Newcastle disease virus and vaccine virus, and identifying its virulence and genotype and its supporting automatic analysis software. Background technique [0002] Newcastle disease (ND) is a severe infectious disease of poultry. Due to its rapid spread, widespread prevalence, high morbidity and mortality, it has caused serious losses to the poultry industry, and countries where Newcastle disease broke out have banned and restricted international trade in poultry products, causing greater losses to the poultry industry. The International Office of Epizootics lists it as a class A infectious disease, my country lists it as a first-class epidemic prevention object, and the inspection and quarantine system lists it as a first-class quarantine object for port quarantine. [0003] Newcastle disease (ND) is an acute infectious disease caused by Newcastle disease...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 单松华邵朝纲徐秀芳徐朝哲张念慈
Owner HUZHOU TEACHERS COLLEGE
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