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A Universal Nest-PCR Primer Set for Amplifying the gp90 Gene of Major Classical Strains of Equine Infectious Anemia Virus

A technology of equine infectious anemia and classic strains, applied in the field of equine infectious anemia virus detection

Inactive Publication Date: 2011-12-14
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

This high variability makes the primers designed to amplify the gp90 gene based on the gene background of a representative strain may not be suitable for gene amplification of other strains

Method used

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  • A Universal Nest-PCR Primer Set for Amplifying the gp90 Gene of Major Classical Strains of Equine Infectious Anemia Virus
  • A Universal Nest-PCR Primer Set for Amplifying the gp90 Gene of Major Classical Strains of Equine Infectious Anemia Virus

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Embodiment 1

[0016] 1 Experimental materials and methods

[0017] 1.1. Virus strains, disease materials, and test samples

[0018] EIAV samples used to verify the specificity of primer amplification include EIAV North American representative strain EIAV wyo (Genbank No, AF033820), South American representative strain EIAV Argen (Recorded in: Geng Qinghua. The establishment of a differential diagnosis method for equine infectious anemia attenuated vaccine strains in my country and the epidemic strains in the Americas and the research on labeled vaccines. Chinese doctoral dissertation, Beijing, Graduate School of Chinese Academy of Agricultural Sciences, 2006.), Chinese representative strain EIAV LN40 (Genbank No, AF327877), Chinese EIAV vaccine attenuated strain EIAV DLV125 (Genbank No, AF327878) and local isolates EIAV-1 and EIAV-2 in China (preserved by the laboratory of the patent applicant and isolated from Heilongjiang in 2004 and 2005, respectively).

[0019] 1.2 Primer design and ...

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Abstract

The invention discloses a general nest-PCR primer set for amplifying the gp90 gene of the main classical strain of equine infectious anemia virus, and also relates to the method of using the primer set to perform nest-PCR amplification on a sample to be tested and detecting equine infectious anemia virus The method and the use of the primer set in preparing biological reagents for detecting or diagnosing equine infectious anemia virus. The invention is based on the whole gene sequence alignment of EIAV main classical strains EIAVwyo, EIAVArgen, EIAVLN40, and designs amplification primers based on the conserved regions. Experimental results show that the nest-PCR primers designed by the present invention can be generally used for the amplification of the gp90 gene of the main representative strain of EIAV, and also have amplification specificity for the field isolates. The nest-PCR primers established in this study can be used in clinical molecular epidemiological studies of EIAV epidemic strains, and are effective general primers for amplifying the gp90 gene.

Description

technical field [0001] The invention relates to detection primers for equine infectious anemia virus, in particular to a general-purpose nest-PCR primer set designed based on nest-PCR for detecting the gp90 gene of the main classic strain of equine infectious anemia virus, and its detection method and application, belonging to equine infectious The field of detection of anemia virus. Background technique [0002] Equine infectious anemia virus (Equine infectious anemia virus, EIAV) is a member of the retroviridae lentivirus genus, and is the pathogen of equine infectious anemia (Equine infectious anemia, EIA), an infectious disease that seriously harms equine animals. [0003] The Gp90 gene is a viral gene encoding the EIAV envelope protein Gp90, with a total length of about 1.4kb. The envelope protein Gp90, as a ligand, interacts with the host cell receptor ELR to help the virus enter the host cell. Due to the poor fidelity of the reverse transcriptase of the lentivirus, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 马建周建华林跃智王雪峰杜承李利
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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