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Kit capable of quickly detecting classical swine fever virus/porcine reproductive and respiratory syndrome virus/pseudorabies virus/porcine parvovirus

A technique for porcine pseudorabies virus and respiratory syndrome, which is applied in the directions of microorganism-based methods, microorganism determination/inspection, microorganisms, etc., which can solve the problems of easy mixed infection and difficulty in diagnosing porcine reproductive disorders, and achieve the time required for detection. The effect of shortening, easy operation and improving detection efficiency

Active Publication Date: 2016-07-06
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because CSFV, PRRSV, PRV and PPV have similar clinical symptoms after infection and are easy to cause mixed infection, it brings certain difficulties to the diagnosis of swine reproductive disorders

Method used

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  • Kit capable of quickly detecting classical swine fever virus/porcine reproductive and respiratory syndrome virus/pseudorabies virus/porcine parvovirus
  • Kit capable of quickly detecting classical swine fever virus/porcine reproductive and respiratory syndrome virus/pseudorabies virus/porcine parvovirus
  • Kit capable of quickly detecting classical swine fever virus/porcine reproductive and respiratory syndrome virus/pseudorabies virus/porcine parvovirus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The preparation of embodiment 1 kit and its use

[0031] 1. Prepare a kit including the following components

[0032] 1 tube of primer-probe mixture (25 μl / tube), 1 tube of RT-PCR reaction solution (250 μl / tube), 1 tube of RT-PCR enzyme system (75 μl / tube), 1 tube of positive quality control (200 μl / tube), Negative quality control (200μl / tube) 1 tube, DEPCH 2 O (2000μl / tube) 1 tube.

[0033] 2. Preparation of target gene recombinant plasmid

[0034] The entrusted company synthesized and purified the recombinant plasmids of 4 kinds of viral target genes, measured the concentration by ultraviolet spectrophotometer, and then calculated the DNA copy number, and used ddH for 4 kinds of DNA 2 O were diluted to 1.0 × 10 5 copies / mL~1.0×10 2 copies / mL of the standard.

[0035] 3. Real-time fluorescent quantitative PCR amplification and detection

[0036] 3.1 Reagent preparation:

[0037] Take the corresponding amount of primer-probe mixture 1 μl, RT-PCR reaction solutio...

Embodiment 2

[0045] Example 2 Application of a rapid detection of classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus, and porcine parvovirus kit to detect specific samples

[0046] Select clinically confirmed positive samples of foot-and-mouth disease virus, porcine circovirus type 2, rabies virus, Erysipelas suis, Leptospira, porcine encephalitis B virus, and swine influenza virus as specific samples. Nucleic acid extraction, PCR amplification and result analysis steps were carried out with reference to Example 1, and the detection of negative and positive quality control products was carried out at the same time.

[0047] Test result: the amplification curve of the negative quality control product is not S-shaped (see attached figure 1 ), the amplification curve of the positive quality control product is an obvious S-shaped curve (see attached figure 2 , 3, 4, 5), both negative and positive quality control products meet the qual...

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Abstract

The invention relates to a kit capable of quickly detecting classical swine fever virus / porcine reproductive and respiratory syndrome virus / pseudorabies virus / porcine parvovirus, in particular to a multiplex real-time fluorescence polymerase chain reaction technology capable of detecting CSFV (classical swine fever virus), PRRSV (porcine reproductive and respiratory syndrome virus), PRV (pseudorabies virus) and PPV (porcine parvovirus) simultaneously.The kit mainly comprises a RT-PCR (reverse transcription-polymerase chain reaction) mixture, primer probe mixed liquor, an RT-PCR enzyme system and DEPC (diethyl pyrocarbonate) H2O as well as packaging boxes for packaging reagent bottles or tubes separately and in a centralized manner.Through multiple fluorescence channels for separate detection, the kit applying a one-step real-time fluorescence PCR mode is capable of detecting and identifying the classical swine fever virus, the porcine reproductive and respiratory syndrome virus, the pseudorabies virus and the porcine parvovirus quickly and accurately and can be widely applied to multiple fields such as early clinical diagnosis of porcine reproductive disturbance diseases, port inspection and quarantine, plague prevention and scientific research.

Description

technical field [0001] The invention relates to a rapid detection kit for classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus, and porcine parvovirus, in particular to a kit that utilizes multiple real-time fluorescent polymerase chain reaction techniques to react in one tube Kits for detecting and identifying classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus, and porcine parvovirus. Background technique [0002] Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (Porcinereproductiveandrespiratorysyndromevirus, PRRSV), porcine pseudorabies virus (Pseudorabiesvirus, PRV) and porcine parvovirus (Porcineparvovirus, PPV) are the four major causes of reproductive disorders in pigs. pathogens of infectious diseases. In large-scale and intensive pig farms, reproductive disorders have become one of the most important diseases and h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6851C12Q1/701C12Q1/705C12Q2600/16C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/113
Inventor 李明何皖平周其伟蒋析文高秀洁周阿涛李丽梅
Owner DAAN GENE CO LTD
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