Method for detecting feline herpes virus

A feline herpes virus and purpose technology, applied in the field of molecular detection, can solve the problems of low virus resistance, low sensitivity, and low success rate, and achieve the effects of low cost and high sensitivity

Inactive Publication Date: 2017-07-25
QINGDAO AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, in terms of etiological diagnosis, due to the relatively large workload of isolation and culture, and the low resistance of the virus itself to the external environment, the success rate of isolation is not very high
The most rapid and convenient virus electron microscope identification after isolation and culture can

Method used

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  • Method for detecting feline herpes virus
  • Method for detecting feline herpes virus
  • Method for detecting feline herpes virus

Examples

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Embodiment 1

[0030] Example 1: Design and screening of primers

[0031] The FHV-1C-27 strain gene (sequence FJ478159) registered in GenBank was compared by MegLign in DNAStar software, using Primer Premier 5.0 software, PCR primers and Taq man probe sequences, using fluorescent groups as the luminescent groups of the probes , using different wavelength bands of the corresponding fluorescent quantitative PCR instrument for detection. Design and synthesize a pair of FHV specific primers P1 / P2. The designed primers were analyzed as follows: interference between primers, hairpin structure and dimer. And through the BLAST function of NCBI to preliminarily identify its specificity. After screening, the primers with complementary sequences between the primers themselves and between the primers, and the single strand of the amplification product that can form a secondary structure were excluded, and the primers and probe sequences shown in Table 1 below were selected. The primers were synthesiz...

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Abstract

The invention provides a method for detecting feline herpes virus. The method comprises the following steps: screening to obtain an oligonucleotide primer, and detecting feline herpes virus by adopting a SYBR Green I fluorescent quantitative PCR method. The invention provides a primer pair for detecting feline herpes virus, and the sequences of an upstream primer and a downstream primer are respectively SEQ ID NO:1 and SEQ ID NO:2. The screened primer not only can ensure the accuracy of identified pathogen, but also can not detect congener different strains of pathogen. In addition, the amplified segments have a proper length, thus not only being capable of separating from each other in electrophoresis, but also being incapable of dispersing, and avoiding the situation that small segments are optimally amplified and the amplification of long segments are blocked in multiple PCR; and non-specific amplified products such as a primer dimer and the like. in addition, the high-specificity and high-sensitivity fluorescent quantitative PCR detection method for FHV can be obtained by the optimization of a fluorescent quantitative reaction system and reaction conditions.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and in particular relates to a method for detecting feline herpes virus, that is, the feline herpes virus is detected by SYBR Green I fluorescent quantitative PCR method using oligonucleotide primers obtained by screening. Background technique [0002] Feline herpes virus (FHV, feline rhinotracheitis), the virus mainly affects young cats, and can cause feline rhinotracheitis disease. When the symptoms of infected cats are severe, there will be elevated body temperature and obvious upper respiratory tract infection. , Throat, trachea and bronchi, tongue, conjunctiva, etc. proliferate. The disease was first discovered in the United States, and then found and spread in Canada, the United Kingdom, the Netherlands, Switzerland, Hungary, Vietnam and other places. Suspicious cases have been found many times in my country and the virus has been isolated. However, there are currently no testi...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/705C12Q2531/113C12Q2563/107
Inventor 单虎潘柳婷倪彬砚张慧秦志华张洪亮杨瑞梅于鹏孙军昌孙强
Owner QINGDAO AGRI UNIV
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