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

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

Active Publication Date: 2020-06-19
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • 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 for African swine fever virus wild strain
  • Locked nucleic acid probe fluorescent quantitative PCR detection composition, detection method and detection kit for African swine fever virus wild strain
  • Locked nucleic acid probe fluorescent quantitative PCR detection composition, detection method and detection kit for 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 locked nucleic acid probe fluorescent quantitative PCR detection composition, a detection method and a detection kit for African swine fever virus wild strain. The detectioncomposition is designed by utilizing characteristics of the locked nucleic acid, has strong recognition capability and affinity to DNA, can effectively improve detection efficiency of the African swine fever virus wild strain, and reduces probability of missed detection. Compared with an existing detection method, the detection method has advantages of strong specificity, high sensitivity and thelike, and is 10 times higher than that of a conventional TaqMan probe fluorescence quantitative PCR method. The detection composition, detection method and detection kit can be used to distinguish between African swine fever virus wild strains and MG360-505R gene-deleted vaccine strains, and are particularly suitable for scientific research and clinical applications, and have 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 Applications(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
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