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Two-color fluorogenic quantitative PCR (Polymerase Chain Reaction) detection kit for CDV (canine distemper viruses) and CPV (canine parvo viruses)

A technology of canine parvovirus and detection kit, which is applied in the biological field, can solve problems such as false positives, non-specific amplification, and insufficient simplification of operation methods, and achieve the effects of simple application, avoiding missed diagnosis and misdiagnosis, and enhancing sensitivity and specificity

Active Publication Date: 2014-03-05
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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Problems solved by technology

However, although the sensitivity of canine distemper and canine parvovirus nucleic acid detection is improved by ordinary PCR method, it is easy to produce non-specific amplification, and even false positives may occur due to operational reasons, and the results of ordinary PCR amplification method are low. It is judged that the product needs to be analyzed by gel electrophoresis, and the operation method is not simplified enough

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  • Two-color fluorogenic quantitative PCR (Polymerase Chain Reaction) detection kit for CDV (canine distemper viruses) and CPV (canine parvo viruses)
  • Two-color fluorogenic quantitative PCR (Polymerase Chain Reaction) detection kit for CDV (canine distemper viruses) and CPV (canine parvo viruses)
  • Two-color fluorogenic quantitative PCR (Polymerase Chain Reaction) detection kit for CDV (canine distemper viruses) and CPV (canine parvo viruses)

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Embodiment 1

[0069] 1. Design of specific primers and probes

[0070] According to the nucleotide sequences of canine distemper virus and canine parvovirus retrieved from Genbank, primers and probes for detecting canine distemper virus and canine parvovirus were designed, and their sequences were as follows:

[0071] Canine Distemper Virus:

[0072] Upstream primer: CDV-F: 5'- GCAAGCGAGGATTGAGGGTAT-3' (SEQ ID NO.1);

[0073] Downstream primer: CDV-R: 5'- GGCCTATGGGGACACACTCAAA-3' (SEQ ID NO.2);

[0074] Fluorescent probe: CDV-P: 5'-FAM-CACCGACCATCCAGACGTTGCTCCT-TAMRA-3' (SEQ ID NO.3), amplified fragment 74 bp;

[0075] Canine parvovirus:

[0076] Upstream primer: CPV-F: 5'-CGTCTACACAAGGGCCATTTA-3' (SEQ ID NO.4);

[0077] Downstream primer: CPV-R: 5'- CCATGTTGTCTACCAAATGCAT-3' (SEQ ID NO.5);

[0078] Fluorescent probe: CPV-P: 5'-VIC-CGCAAACAGATGAAAATCAAGCAGCA-TAMRA-3' (SEQ ID NO.6), amplified fragment of 116 bp.

[0079] The fluorescent probe reporter group used to detect canine dis...

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Abstract

The invention discloses a two-color fluorogenic quantitative PCR (Polymerase Chain Reaction) detection kit for CDV (canine distemper viruses) and CPV (canine parvo viruses). A detection method comprises the following steps: firstly, pre-treating a sample by using a nucleic acid extraction solution of the kit and extracting nucleic acid; then detecting the nucleic acids of the CDV and CPV simultaneously in a reaction system by utilizing the two-color fluorogenic quantitative PCR detection kit, and detecting whether the canine distemper and canine parvo viruses exist in the sample to be detected or not by one step; and quantifying a virus copy number. The detection method is high in detection efficiency and good in specificity and sensibility; the aims of clinical detection and epidemic situation control of animal epidemic diseases can be realized.

Description

technical field [0001] The invention relates to a two-color fluorescent quantitative PCR detection kit, in particular to a two-color fluorescent quantitative PCR detection kit for simultaneously detecting canine distemper and canine parvovirus. The invention belongs to the field of biotechnology. Background technique [0002] Canine distemper virus (Canine distempervirus, CDV) and canine parvovirus (Canine Parvovirus, CPV) are two of the most serious infectious disease pathogens that harm dogs. Severe sequelae, severe can lead to acute death of a large number of dogs. [0003] Canine distemper virus (CDV) is a single-stranded RNA virus whose hosts are canids and mustelids, and is mainly transmitted horizontally through air and droplets. Sick dogs are an important source of infection, and excrete toxins through urine for a long time, polluting the surrounding environment; dogs of different ages, sexes and breeds can be infected. The symptoms of CDV infection are various, wh...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q1/70C12Q2563/107C12Q2531/113
Inventor 魏文康徐贵峰谭婉红林立峰袁洁吕殿红翟少伦温肖会周秀蓉贾春玲陈琴苓
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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