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Canine distemper virus and canine parvovirus duplex TaqMan-MGB fluorescent quantitative PCR (polymerase chain reaction) detection kit and detection method thereof

A canine parvovirus and fluorescence quantitative technology, which is applied in the field of animal disease quarantine and detection, can solve the problems of long test period, difficulty in meeting rapid, large-scale detection, technical requirements and high test conditions.

Active Publication Date: 2014-04-30
HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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Problems solved by technology

Currently available CDV / CPV detection methods mainly include virus isolation and identification, immunofluorescence (IF), immunohistochemistry (IHT), immunoelectron microscopy (IEM), hemagglutination / hemagglutination inhibition test (HA / HI) , neutralization test and enzyme-linked immunosorbent assay (ELISA) and other methods, these methods have the disadvantages of cumbersome operation, high technical requirements and test conditions, long test cycle, etc. A single PCR or FQ-PCR detection method for a virus

Method used

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  • Canine distemper virus and canine parvovirus duplex TaqMan-MGB fluorescent quantitative PCR (polymerase chain reaction) detection kit and detection method thereof
  • Canine distemper virus and canine parvovirus duplex TaqMan-MGB fluorescent quantitative PCR (polymerase chain reaction) detection kit and detection method thereof
  • Canine distemper virus and canine parvovirus duplex TaqMan-MGB fluorescent quantitative PCR (polymerase chain reaction) detection kit and detection method thereof

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Embodiment Construction

[0045] ,Materials and Methods

[0046] 1.1 Materials

[0047] 1.1.1 Virus strains

[0048] The isolated strains of canine distemper and canine parvovirus were isolated and identified by the laboratory of Henan Provincial Center for Animal Disease Prevention and Control; other control virus or bacterial strains were kept by the laboratory of Henan Provincial Center for Animal Disease Prevention and Control;

[0049] 1.1.2 Instruments and reagents

[0050] Fluorescence PCR instrument, product of ABI Company in the United States, model ABI7000; PCR amplification instrument, product of Biometra Company in Germany; gel imaging analysis system, product of Alpha Innotech Company in the United States; constant temperature water bath oscillator (HZQ-Q), developed by Harbin Donglian Electronic Technology Co., Ltd. products; desktop high-speed refrigerated centrifuges, products of Heraeus Company in the United States; Ex Taq DNA polymerase, dNTPs, DNA recovery kit, etc. were...

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Abstract

The invention belongs to the technical field of animal epidemic disease detection and quarantine, and discloses a canine distemper virus (CDV) and canine parvovirus (CPV) duplex TaqMan-MGB fluorescent quantitative PCR (polymerase chain reaction) detection kit and a detection method thereof. The detection kit comprises a pair of specific primers designed based on CDVH genes, a TaqMan-MGB probe with the 5' end marked as FAM, a pair of specific primers designed based on CPVVP2 genes and a TaqMan-MGB probe with the 5' end marked as VIC. The detection method comprises the following steps: designing two pairs of primer sequences and two TaqMan-MGB marked probe sequences, and performing fluorescent labeling; preparing total RNA (ribonucleic acid) and total DNA (deoxyribonucleic acid); performing reverse transcription; performing fluorescent PCR amplification; performing result judgment. According to the detection method, the nucleic acids of a CDV and a CPV can be detected at the same time and the CDV / CPV can be quickly, specifically and sensitively identified at the same time. The detection kit is of great significance on prevention and control of CDV and CPV diseases.

Description

technical field [0001] The invention belongs to the technical field of animal epidemic quarantine and detection. It relates to canine distemper virus and canine parvovirus duplex TaqMan MGB fluorescence quantitative PCR detection reagent and detection method. Background technique [0002] Canine distemper (CD) is an acute, highly contagious infectious disease of dogs that occurs worldwide. Its pathogen is Canine Distemper Virus (CDV), which was first reported in 1905. Its natural hosts include large Some carnivores. The disease is highly contagious, and it is prone to mixed infection or secondary infection of bacteria and other viruses. The incidence rate is as high as 80%. After recovery, sequelae such as paralysis, convulsions, and epileptic seizures are prone to occur. There is no specific drug treatment for this disease, and it has become one of the main diseases that endanger the dog industry. CDV belongs to the genus Morbillivirus of the family Paramyxoviridae, a ne...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q1/70C12Q2537/143C12Q2561/101C12Q2531/113
Inventor 吴志明闫若潜赵明军赵雪丽王淑娟刘毅王东方安春霞谢彩华钱勇刘梅芬张代宝张林海
Owner HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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