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Multiplex polymerase chain reaction (PCR) detection primers for simultaneously detecting canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus

A technology for canine parvovirus and canine distemper virus, applied in the direction of microorganism-based methods, microorganisms, recombinant DNA technology, etc., can solve the problems of difficult diagnosis and long time consumption, and achieve high sensitivity, good repeatability and strong specificity Effect

Inactive Publication Date: 2012-08-01
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

[0004] The clinical symptoms of these major diseases in dogs are similar, and there are often mixed infections, which makes diagnosis difficult
The commonly used single PCR method takes a long time, and it is necessary to establish a rapid early diagnosis method so that effective prevention and control measures can be taken as soon as possible to reduce the harm caused by these diseases to the dog industry

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  • Multiplex polymerase chain reaction (PCR) detection primers for simultaneously detecting canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus
  • Multiplex polymerase chain reaction (PCR) detection primers for simultaneously detecting canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus
  • Multiplex polymerase chain reaction (PCR) detection primers for simultaneously detecting canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus

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Embodiment 1

[0025] 1 Materials and methods

[0026] 1.1 Virus strains CDV strains, CPV strains, CAV-I, CAV-II strains and rabies virus (RV) strains are all preserved by the Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences.

[0027] 1.2 Main reagents

[0028] Ex Taq DNA polymerase, dNTP, DL 2000 DNA Marker, M-MLV enzyme and RNase inhibitor were purchased from Bao Biological Engineering (Dalian) Company; DNA extraction kit was purchased from Omega Company; RNA extraction kit was purchased from Qiagen Company .

[0029] 1.3 Primer design

[0030] According to the CDVN gene sequence, CPVNS gene sequence and CAV E3 gene sequence in GenBank, 3 pairs of specific primers were designed, and the primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. See Table 1 for the sequence.

[0031] Table 1 Multiplex PCR Primer Sequence

[0032]

[0033]

[0034] 1.4 Extraction of RNA and DNA

[0035] Extract viral RNA and viral D...

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Abstract

The invention discloses a set of multiplex polymerase chain reaction (PCR) detection primers for detecting canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus. The primers comprise three pairs of primers, wherein sequences of primers for detecting canine distemper virus are shown as SEQ ID NO:1 and SEQ ID NO:2; sequences of primers for detecting canine parvovirus are shown as SEQ ID NO:3 and SEQ ID NO:4; and sequences of primers for detecting type I canine adenovirus and type II canine adenovirus are shown as SEQ ID NO:5 and SEQ ID NO:6. The primers can simultaneously detect canine distemper virus, canine parvovirus, type I canine adenovirus and type II canine adenovirus through a multiplex PCR method, and have the advantages of high specificity, high sensitivity, high repeatability and the like.

Description

technical field [0001] The invention relates to a set of primers for detecting viruses, in particular to a set of primers for simultaneous detection of canine distemper virus, canine parvovirus, type I and type II canine adenovirus multiple PCR methods, and belongs to the field of virus detection. Background technique [0002] Canine distemper virus (CDV), canine parvovirus (CPV) and canine adenovirus (CAV) are the main pathogens causing canine disease, and these three viruses are clinically fatal after single or mixed infection The rate can reach 50%-100%, causing huge economic losses to the dog industry. [0003] Clinically, canine distemper caused by CDV mainly includes four types: respiratory type, digestive type, neurological type and mixed type. CPV mainly causes acute hemorrhagic gastroenteritis in dogs and acute myocarditis in puppies. CAV can be divided into type 1 and type 2 according to its different characteristics such as hemagglutination and neutralization tes...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 刘大飞张洪英曲连东林欢姜骞
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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