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Taqman probe fluorescent quantitative PCR detection kit for identifying canine distemper virus wild strain and vaccine strain as well as application thereof

A canine distemper virus, fluorescence quantitative technology, applied in the field of medicine, can solve problems such as inconvenient use

Pending Publication Date: 2018-11-20
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods require multiple PCRs and are inconvenient to use
There is no report on the differential diagnosis of CDV using TaqMan probes

Method used

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  • Taqman probe fluorescent quantitative PCR detection kit for identifying canine distemper virus wild strain and vaccine strain as well as application thereof
  • Taqman probe fluorescent quantitative PCR detection kit for identifying canine distemper virus wild strain and vaccine strain as well as application thereof
  • Taqman probe fluorescent quantitative PCR detection kit for identifying canine distemper virus wild strain and vaccine strain as well as application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The establishment of embodiment 1 canine distemper virus wild strain and vaccine strain identification method

[0035] 1 Materials and methods

[0036] 1.1 strain

[0037] CDV virulent HBF-1 strain and SD14(7) strain, vaccine virus CDV3 strain and CDV-OR12 strain, canine parainfluenza virus (CPIV), rabies virus (RABV), canine coronavirus (CcOV), canine parvovirus (CPV) ), canine adenovirus type Ⅰ (CAV-Ⅰ) and canine adenovirus type Ⅱ (CAV-Ⅱ), all of which are preserved by the Disease Prevention and Control Laboratory of the Institute of Special Products, Chinese Academy of Agricultural Sciences.

[0038] 1.2 Main reagents and instruments

[0039] The real-time fluorescent quantitative PCR instrument Light Cycler○R96 was purchased from Roche; the common PCR amplification instrument GeneAmp PCRSystem 2700 and the spectrophotometer Nanodrop 2000 were purchased from Thermo; the plasmid mini-extraction kit was purchased from Axygen; the fluorescent quantitative PCR kit Fast...

Embodiment 2

[0072] Example 2 Assembly of Taqman probe fluorescent quantitative PCR detection kit for distinguishing canine distemper virus wild strain and vaccine strain

[0073] 1. Primers and probes

[0074] Universal primers for amplifying canine distemper virus field strains and vaccine strain H gene conserved regions, the universal primers are composed of upstream primers and downstream primers, the nucleotide sequence of the upstream primers is as SEQ ID NO. 1, the nucleotide sequence of the downstream primer is shown in SEQ ID NO.2;

[0075] A specific probe for identifying canine distemper virus wild strains, its nucleotide sequence is shown in SEQ ID NO.3, the 5' end of the probe is added with a fluorescent group FAM, and the 3' end is added with a quencher Killing group TAMRA;

[0076] A specific probe for identifying canine distemper virus vaccine strains, its nucleotide sequence is shown in SEQ ID NO.4, the 5' end of the probe is added with a fluorescent group FAM, and the 3...

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Abstract

The invention discloses a Taqman probe fluorescent quantitative PCR detection kit for identifying a canine distemper virus wild strain and a vaccine strain as well as application thereof. The kit comprises a universal primer for amplifying a canine distemper virus wild strain and vaccine strain H gene conservation area as well as a specific probe for identifying a canine distemper virus wild strain or a vaccine strain. The experiment proves that the Taqman probe fluorescent quantitative PCR detection kit is used for detecting and identifying the CDV wild strain and the vaccine strain, the sensitivity can reach to 1 copy; 103 to 102 copies with the virus quantity as low as 1 TCID50 / mL and higher than that of the conventional method can be detected; and through detection on five kinds of other pathogen nucleic acid, the copies are negative. Therefore, the kit for identifying the CDV wild strain and the vaccine strain has the characteristics of high specificity, high sensitivity and highstability, and can rapidly identify canine distemper virus wild strain and vaccine strain infection.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR detection kit and its application, in particular to a Taqman probe fluorescent quantitative PCR detection kit for distinguishing canine distemper virus wild strains and vaccine strains and its application. The invention belongs to the technical field of medicine. Background technique [0002] Canine distemper (Canine Distemper) is an acute febrile, highly contagious infectious disease caused by canine distemper virus (Canine Distemper Virus) infecting carnivores such as dogs, fox raccoons and minks. The disease has a high morbidity rate and various clinical symptoms. It is prone to secondary mixed infection in the later stage of infection, and the mortality rate is as high as 30%-100%. In recent years, the number of CDV natural infection hosts has continued to increase. With the increase in the number of pets such as dogs and cats, canine distemper virus has brought great losses and harm to the a...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2531/113C12Q2563/107
Inventor 薛向红闫喜军闫鸣昊刘建雨卜研史宁朱言柱廉士珍张蕾胡博
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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