Fluorescence quantitative PCR kit for quantitatively detecting chicken Marek's virus Meq gene deletion vaccine strain and wild strain and application thereof

A gene deletion vaccine, Marek's technology, applied in the field of biomedicine

Active Publication Date: 2019-02-19
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective and accurate fluorescent quantitative method for distinguishing MDV gene deletion vaccines from wild strains

Method used

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  • Fluorescence quantitative PCR kit for quantitatively detecting chicken Marek's virus Meq gene deletion vaccine strain and wild strain and application thereof
  • Fluorescence quantitative PCR kit for quantitatively detecting chicken Marek's virus Meq gene deletion vaccine strain and wild strain and application thereof
  • Fluorescence quantitative PCR kit for quantitatively detecting chicken Marek's virus Meq gene deletion vaccine strain and wild strain and application thereof

Examples

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Effect test

Embodiment 1

[0042] Embodiment 1 Quantitative detection of chicken Marek's virus Meq gene deletion vaccine strain and the establishment of the fluorescent quantitative PCR method of wild strain

[0043] 1. Primer and probe design

[0044] It can only detect chicken Marek's disease virus Meq gene deletion vaccine strain rMSΔMeq strain (this strain is recorded in the patent application whose publication number is CN102363769B, and the title of invention is "Chicken Marek's disease virus Meq gene deletion vaccine strain, its construction method and application" , its microorganism preservation number is: CGMCC No.4612) the fluorescent quantitative PCR is named: rMS-qPCR, the upstream and downstream primers and probes of rMS-qPCR are designed as follows figure 1 Shown; The fluorescent quantitative PCR that detects all MDV wild strains is named: MDV-qPCR, the upstream and downstream primers and probes of MDV-qPCR are designed as follows figure 2 shown. The sequences of all primers and probes...

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Abstract

The invention discloses a fluorescence quantitative PCR kit for quantitatively detecting chicken Marek's virus Meq gene deletion vaccine strain and wild strain and an application thereof. The kit comprises two sets of fluorescent quantitative PCR primers and probes respectively used for detecting the chicken Marek's virus Meq gene deletion vaccine strain and wild strain, wherein a sequence of thefluorescent quantitative PCR primer used for detecting the chicken Marek's virus Meq gene deletion vaccine strain is shown in SEQ ID NO. 1 and SEQ ID NO.2, the sequence of the probe is shown in SEQ IDNO. 3; the sequence of the fluorescent quantitative PCR primer used for detecting the wild strain of the chicken Marek's virus is shown in SEQ ID NO. 4 and SEQ ID NO.5, and the sequence of the probeis shown in SEQ ID NO. 6. Vitro experiments prove that the kit has high sensitivity, good specificity and good repeatability, can respectively and accurately quantify the chicken Marek's virus Meq gene deletion vaccine strain and MDV wild strain, and has important significance for diagnosis of the chicken Marek's disease, immune monitoring of the vaccine and the like.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR kit and an application thereof, in particular to a fluorescent quantitative PCR kit for quantitatively detecting chicken Marek's virus Meq gene deletion vaccine strains and wild strains and the application thereof. The invention belongs to the technical field of biomedicine. Background technique [0002] Chicken Marek's disease (Marek's Disease, MD) is an important viral infectious disease of chickens, and the pathogen is chicken Marek's disease virus (Marek's Disease virus, MDV). MD epidemiological studies have confirmed that wild MDV strains in my country continue to mutate and evolve toward stronger virulence. MDV virulent strains that can break through the protection of existing MD vaccines (CVI988 and 814 strains) have emerged. [0003] Aiming at the increasing virulence of MDV and seriously endangering the development of the poultry industry, Chinese scholars have developed a chicken Marek'...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/6851C12Q1/705C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 刘长军王笑梅张艳萍李凯高玉龙崔红玉高宏雷
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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