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Detection primer, detection kit and virus detection method for fowl adenovirus-I, and application of primer

A technology of avian adenovirus and detection primers, which is applied in the field of detection primers for group I avian adenoviruses, and can solve problems such as non-specificity and limitations of detection methods.

Pending Publication Date: 2019-12-10
CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the disease can be preliminarily diagnosed based on clinical manifestations and pathological autopsy, and confirmed by virus isolation and culture, histopathological observation and PCR identification, but the existing detection methods have certain limitations and non-specificity

Method used

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  • Detection primer, detection kit and virus detection method for fowl adenovirus-I, and application of primer
  • Detection primer, detection kit and virus detection method for fowl adenovirus-I, and application of primer
  • Detection primer, detection kit and virus detection method for fowl adenovirus-I, and application of primer

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Effect test

Embodiment 1

[0112] Embodiment 1: the sensitivity test of kit of the present invention detects FAdV-Ⅰ

[0113] (1) Sample: FAdV-I non-labeled positive control nucleic acid.

[0114] (2) Sample treatment: Dilute the FAdV-I non-labeled positive control nucleic acid to 1000, 100, 10, 1, 0.1 pg / μl, and negative control nucleic acid on the nylon membrane in sequence, mark and perform dot hybridization .

[0115] (3) Use tweezers to place the nylon membrane (with the sampling side up) on the filter paper saturated with denaturing solution (the filter paper should be placed in a glass plate in advance) for denaturation for 10 minutes, then take it out with tweezers and place it in the filter paper saturated with neutralizing solution. Neutralize for 5 minutes on a filter paper (the filter paper is placed in a glass plate in advance).

[0116] (4) Take out the film with tweezers, put it on a plate (with the sample point facing up) and dry it at room temperature for 10 minutes until there are no ...

Embodiment 2

[0140] Embodiment 2: The kit of the present invention detects the specificity test of FAdV-Ⅰ

[0141] (1) Samples: Avian Reticuloendothelioma Virus (REV) Genomic DNA, Chicken Marek's Disease Virus (MDV) Genomic DNA, Chicken Infectious Anemia Virus (CIAV) Genomic DNA, Chicken Leukemia Virus (ALV) Genomic DNA, Chicken Reovirus (ARV) Genomic DNA, Chicken Infectious Bronchitis Virus (IBV) Genomic DNA, Parrot Beaking Disease Virus (PBFDV) Genomic DNA, Avian Polyoma Virus (APV) Genomic DNA DNA.

[0142] (2) Sample processing: Take 1 μl of the negative control nucleic acid and positive control nucleic acid in the kit and 2 μl of the genomic DNA of REV, MDV, CIAV, ALV, ARV, IBV, PBFDV and APV respectively, and spot them on the nylon membrane respectively. on, mark it.

[0143] (3) Use tweezers to place the nylon membrane (with the sampling side up) on the filter paper saturated with denaturing solution (the filter paper should be placed in a glass plate in advance) for denaturation ...

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Abstract

The invention belongs to the technical field of animal pathogen molecule detection, and particularly relates to a detection primer, a detection kit and a virus detection method for a fowl adenovirus-I, and application of the primer. A specific primer is used for synthesizing a digoxigenin-labeled specific nucleic acid probe through PCR (Polymerase Chain Reaction). Through dot blot, the specific probe can be used for detecting the existence of 12 types of serotype nucleic acids of the fowl adenovirus-I in a pathological material sample, and is used for judging whether fowl adenovirus-I infection is in the presence or not. By use of the kit to which the invention relates, the DNA (deoxyribonucleic acid) of a pathological material tissue sample is extracted to carry out the dot blot, detection can be finished in 24-36h, a result is reported, so that specificity is high, and accuracy is high.

Description

technical field [0001] The invention belongs to the technical field of molecular detection of animal pathogens, and in particular relates to a group I poultry adenovirus detection primer, a detection kit, a virus detection method and an application. Background technique [0002] Fowl adenovirus (FAdV) is a common infectious pathogen of poultry. The virus is an opportunistic pathogen in poultry, replicating in healthy animals without causing disease or causing only mild clinical symptoms. But adenoviruses can become highly pathogenic when animals are stressed or co-infected with other diseases. FAdV is divided into three groups, among which group I FAdV (FAdV-I) is isolated from the respiratory tract infection of chickens, geese and ducks, and has 5 species and 12 serotypes in total. FAdV-I mainly causes poultry hydropericardium, inclusion body hepatitis, aplastic anemia, hemorrhage, growth retardation, mild respiratory disease, and decreased egg production in laying hens. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6834C12N15/11
CPCC12Q1/701C12Q1/6834C12Q2600/166C12Q2531/113C12Q2545/113
Inventor 李阳蒋文明于晓慧王静静周晓翠王琳刘华雷
Owner CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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