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Multiplex PCR (polymerase chain reaction) primer group for simultaneously detecting feline panleucopenia virus, feline calicivirus and feline herpes virus type 1

A technology for leukopenia and feline calicivirus, applied in recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve problems such as time-consuming and difficult diagnosis, and achieve high sensitivity and specificity , good repeatability

Inactive Publication Date: 2013-09-04
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

[0005] The clinical symptoms of these major diseases in cats are similar, and often have 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

Method used

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  • Multiplex PCR (polymerase chain reaction) primer group for simultaneously detecting feline panleucopenia virus, feline calicivirus and feline herpes virus type 1
  • Multiplex PCR (polymerase chain reaction) primer group for simultaneously detecting feline panleucopenia virus, feline calicivirus and feline herpes virus type 1
  • Multiplex PCR (polymerase chain reaction) primer group for simultaneously detecting feline panleucopenia virus, feline calicivirus and feline herpes virus type 1

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Embodiment 1

[0020] 1 Materials and methods

[0021] 1.1 Virus strains

[0022] FPV strains, FCV strains, FHV1 strains and rabies RV strains were all preserved by the Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences.

[0023] 1.2 Main reagents

[0024] Ex Taq DNA polymerase, dNTP, DL2000 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.

[0025] 1.3 Primer design

[0026] According to the FPV NS gene sequence, FHV TK gene sequence and FCV ORF2 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.

[0027] Table 1 Multiplex PCR Primer Sequence

[0028]

[0029] 1.4 Extraction of RNA and DNA

[0030] Extract viral RNA and viral DNA according to the i...

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Abstract

The invention discloses a multiplex PCR (polymerase chain reaction) primer group for simultaneously detecting feline panleucopenia virus, feline calicivirus and feline herpes virus type 1. The primer group comprises three pairs of primers, wherein the sequences of the primers for detecting the feline panleucopenia virus are shown in SEQ ID NO:1 and SEQ ID NO:2, the sequences of the primers for detecting the feline calicivirus are shown in SEQ ID NO:3 and SEQ ID NO:4, and the sequences of the primers for detecting the feline herpes virus type 1 are shown in SEQ ID NO:5 and SEQ ID NO:6. The feline panleucopenia virus, feline calicivirus and feline herpes virus type 1 can be simultaneously detected by using the primer sequences through a multiplex PCR method, the result is positive, while the detection result of other common feline viruses is negative, which shows that the primer group has the advantages of strong specificity, high sensitivity, good 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 multiplex PCR primers for simultaneously detecting feline panleukopenia virus, feline calicivirus and feline herpesvirus type 1, and belongs to the field of virus detection. Background technique [0002] Feline panleukopenia virus (Feline panleukopenia virus, FPV), also known as feline distemper virus, feline infectious enteritis virus, and feline parvovirus, belongs to the parvovirus genus of the family Parvoviridae and was first successfully isolated by Bilin in 1957. FPV causes an acute, highly contagious infectious disease in felines characterized by high fever, vomiting, dehydration, severe leukopenia, and hemorrhagic enteritis. The morbidity and mortality are high, and it is often endemic, and recovery animals may also Long-term poisoning will bring serious harm to cats. [0003] Feline calicivirus (FCV) belongs to the family Caliciviridae and a member of ...

Claims

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Application Information

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