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Locked nucleic acid probe fluorescent quantitative PCR detection composition, detection method and detection kit of African swine fever virus wild strain

A technology of African swine fever virus and nucleic acid-locked probes, which is applied in the field of molecular biology, can solve problems such as false positive results, harsh experimental conditions, and cumbersome ELISA operations, and achieve strong specificity, high sensitivity, and improved detection efficiency. Effect

Active Publication Date: 2022-03-29
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The operation of virus isolation and ELISA is cumbersome and the experimental conditions are harsh, so it is difficult to carry out in ordinary laboratories
Fluorescent quantitative PCR has been widely used in the detection of ASFV, but it has the disadvantage of false positive results due to non-specific amplification

Method used

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  • Locked nucleic acid probe fluorescent quantitative PCR detection composition, detection method and detection kit of African swine fever virus wild strain
  • Locked nucleic acid probe fluorescent quantitative PCR detection composition, detection method and detection kit of African swine fever virus wild strain
  • Locked nucleic acid probe fluorescent quantitative PCR detection composition, detection method and detection kit of African swine fever virus wild strain

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Embodiment Construction

[0037] The embodiments of the present invention will be described in detail below in conjunction with the embodiments and drawings, but those skilled in the art will understand that the following embodiments and examples are only for illustrating the present invention, and should not be considered as limiting the scope of the present invention. If the specific conditions are not specified, follow the general conditions or the conditions suggested by the manufacturer. The reagents or instruments used were not indicated by the manufacturer, and they were all conventional products that could be purchased from the market.

[0038] Materials, Methods and Results

[0039] 1. Virus strain

[0040] Classical swine fever virus, porcine circovirus type 2, and porcine pseudorabies virus were all purchased from commercial vaccines on the market. Among them, the live vaccine of swine fever virus was purchased from Guangdong Yongshun Biopharmaceutical Co., Ltd., and the live vaccine of por...

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Abstract

The invention discloses a lock nucleic acid probe fluorescence quantitative PCR detection composition, detection method and detection kit of African swine fever virus wild strain. The detection composition of the present invention is designed by utilizing the characteristics of locked nucleic acid, has strong recognition ability and affinity for DNA, can effectively improve the detection efficiency of African swine fever virus wild strains, and reduce the probability of missed detection. The present invention provides a new detection method and detection reagent for the identification and detection of wild strains of African swine fever virus. The sensitivity of the method is 10 times higher; it can be used to distinguish the wild strain of African swine fever virus from the vaccine strain with MGF360-505R gene deletion, especially suitable for scientific research and clinical application, and has good commercial application value.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a lock nucleic acid probe fluorescent quantitative PCR detection composition, a detection method and a detection kit of a wild strain of African swine fever virus. Background technique [0002] African swine fever (ASF) is an acute, severe and highly contagious disease of domestic pigs and wild boars caused by African swine fever virus (ASFV), and the death rate of domestic pigs can reach 100 %. The World Organization for Animal Health (OIE) lists it as a legally notifiable animal disease, and China lists it as a first-class animal disease. [0003] The patent "Attenuated African swine fever virus with gene deletion and its application as a vaccine" (publication number: CN110093324A) has published the African swine fever Pig / CN / HLJ / 2018 Chinese epidemic strain MGF360-505R gene deletion or CD2V and MGF360- The joint deletion of 505R gene can provide 100% immune protection, and has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2531/113C12Q2561/101C12Q2545/114
Inventor 李艳李春玲勾红潮孙铭飞卞志标蔡汝健蒋智勇宋帅楚品品
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI