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Nanometer PCR detection kit for rapidly identifying and diagnosing virulent virus and attenuated virus of porcine pseudorabies virus, and applications thereof

A detection kit, a technology for porcine pseudorabies, applied in the field of PCR detection kits and nano-PCR detection kits, can solve the problems of long time, missed detection, cumbersome and other problems

Inactive Publication Date: 2013-07-31
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a novel nano-PCR detection kit for rapid detection and identification of attenuated and wild strains of porcine pseudorabies virus (PRV), so as to solve the time-consuming and cumbersome problems of using existing kits for detection. Shortcomings, and overcome the latent infectivity of PRV, sometimes the problem of missed detection

Method used

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  • Nanometer PCR detection kit for rapidly identifying and diagnosing virulent virus and attenuated virus of porcine pseudorabies virus, and applications thereof
  • Nanometer PCR detection kit for rapidly identifying and diagnosing virulent virus and attenuated virus of porcine pseudorabies virus, and applications thereof
  • Nanometer PCR detection kit for rapidly identifying and diagnosing virulent virus and attenuated virus of porcine pseudorabies virus, and applications thereof

Examples

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Effect test

Embodiment 1

[0042] Example 1. Preparation of a novel nano-PCR detection kit for rapid identification and diagnosis of strong and weak porcine pseudorabies virus (PRV)

[0043] The components of the kit of the present invention include: 2×Nano PCR buffer (purchased from Shandong Dazheng Medical Instrument Co., Ltd.), DNA polymerase (purchased from Shandong Dazheng Medical Instrument Co., Ltd.), primer mixture (10×), positive template The mixture (composed of PRV-pMD18-T-gB, PRV-pMD18-T-gE and PRV-pMD18-T-gG in equimolar ratio, among them, PRV-pMD18-T-gE is used as the wild strain positive control Plasmid, PRV-pMD18-T-gG as positive control plasmid for gE-deficient vaccine strain, PRV-pMD18-T-gB as positive control plasmid for wild strain and gE-deletion vaccine strain), dimethyl sulfoxide (DMSO ), without nucleic acid ddH 2 O.

[0044] The primer mixture described therein is composed of 3 pairs of primers according to an equimolar ratio, the sequences of the first pair of primers are sho...

Embodiment 2

[0054] Example 2. Application of the kit of the present invention in rapid identification and diagnosis of strong and weak porcine pseudorabies virus (PRV)

[0055] 1. Method

[0056] (1) Extract the DNA of the tissue sample to be tested;

[0057] (2) The distribution ratio of each group of the detection system is as follows:

[0058] Nano-PCR reaction solution 10.8μL

[0059] Template mixture or virus DNA to be tested 1.2μL

[0060] A positive control (template mixture is a positive control) and a nucleic acid-free ddH without template are required for each sample to be tested. 2 O served as a negative control.

[0061] 10.8 μL nano-PCR reaction solution includes: 6 μL of 2×Nano PCR buffer, 0.5 μL of Taq DNA polymerase (10 U / μL), 1.2 μL of primer mixture (10×, 100 μmol / L), 0.5 μL of dimethyl sulfoxide (DMSO) μL, nucleic acid-free ddH 2 O2.6 μL.

[0062] The optimal amplification conditions for the nano-PCR reaction in a 12 μl reaction system are:

[0063] Pre-denatura...

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Abstract

The invention discloses a nanometer PCR detection kit for rapidly identifying and diagnosing virulent virus and attenuated virus of porcine pseudorabies virus, and applications thereof. The kit comprises a 2*Nano PCR buffer, Taq DNA polymerase, a primer mixture, DNA polymerase, and nucleic acid-free ddH2O. In order to achieve a good detection effect, the kit further can contain a template mixture and a stabilizer. With the kit of the present invention, disadvantages of long time consuming, tedious property and low sensitivity of the existing kit are solved, sensitivity is improved, and early stage and rapid virus detection is achieved. The kit of the present invention has characteristics of strong specificity, high sensitivity and the like, is suitable for early stage and rapid diagnosis, and can be provided for identifying gene deletion strains and wild strain infection. In addition, the kit can be widely used in grassroots veterinary detection institutions, and can provide important significance for PRV detection, diagnosis and identifying diagnosis in large-scale pig farms.

Description

technical field [0001] The invention relates to a PCR detection kit for detecting the strong and weak virus of porcine pseudorabies virus and its application, in particular to a nano-PCR detection kit for rapidly identifying and diagnosing the strong and weak virus of porcine pseudorabies virus and its application. The invention belongs to The field of preventive veterinary testing. Background technique [0002] Pseudorabies virus (Porcine pseudorabies virus, PRV) can cause a variety of diseases in domestic and wild animals with fever, itching (except pigs) and encephalomyelitis as the main symptoms. Pigs are the storage host and source of infection of PRV, which mainly cause abortion, stillbirth and mummification in pregnant sows; newborn piglets are mostly acute lethal type, with obvious neurological symptoms, and the mortality rate is almost 100%; adult pigs are mostly latent infection. Porcine pseudorabies is found in almost all major swine production regions of the wo...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 崔尚金仇铮
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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