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Immune PCR reagent kit for detecting avian leukemia virus

An avian leukemia virus and kit technology, applied in the field of biotechnology detection, to achieve the effect of filling the technical gap

Inactive Publication Date: 2016-07-20
YANGZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since there is currently no effective vaccine or anti-ALV drug for ALV, detection and purification are the main measures to control ALV

Method used

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  • Immune PCR reagent kit for detecting avian leukemia virus
  • Immune PCR reagent kit for detecting avian leukemia virus
  • Immune PCR reagent kit for detecting avian leukemia virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0016] 1) Biotin labeling of monoclonal antibodies

[0017] First, a protein purification column proteinG was used to secrete two monoclonal antibodies against the P27 protein of avian leukemia virus (hybridoma cell line ALVP27-5D3 (accession number: CGMCC No. 7106) and ALVP27-4F12 (accession number: CGMCC No. 7107). Avian leukemia virus P27 protein specific monoclonal antibodies 5D3 and 4F12.)

[0018] Purify (e.g. figure 2 ), and then biotin-labeled the two monoclonal antibodies according to the instructions of Biotin-XXMicroscaleProteinLabelingKit, and dialyzed with PBS (PH7.4) at 4°C to remove free biotin. The hybridoma cell lines ALVP27-5D3 (deposit number: CGMCC No. 7106) and ALVP27-4F12 (deposit number: CGMCC No. 7107) have been disclosed in CN103163299A.

[0019] 2) Preparation of antibody oligonucleotide probe

[0020] Proximity ligation technology KIT (TaqMan) marked with 3'and 5'oligonucleotides of streptomycin (purchased from ThermoFish Scientific), dilute the two antib...

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Abstract

The invention belongs to the field of biological technical detection and particularly relates to an immune PCR reagent kit for detecting avian leukemia virus. The reagent kit comprises two antibody oligonucleotide probes, a coupled reaction solution, DNA ligase, protease, a Fast Master mixed solution and a Universal q PRC reaction solution. The immune PCR reagent kit uses a biotin-labeled avian leukemia virus preventing special antibody and a commercialized ortho-position connecting reagent, can detect avian leukemia virus antigen through immune PCR and can quickly detect the avian leukemia virus antigen in a high-throughout mode through the immune PCR without carrying out a complex nucleic acid extracting process on a sample. The immune PCR reagent kit for detecting the avian leukemia virus can be applied to clinical detection and purification of avian leukemia virus and will fill domestic and oversea correlated technique blank.

Description

Technical field [0001] The invention belongs to the field of biotechnology detection, and specifically relates to an immune PCR kit for detecting avian leukemia virus. Background technique [0002] Avian leukemia virus belongs to the retroviral family. According to its antigenic characteristics and host range, chicken ALV can be divided into 6 subgroups, including A, B, C, D, E and J subgroups. Among them, A, B, C, D and J subgroups are exogenous viruses, and E subgroups are endogenous viruses. Avian leukemia is mainly spread vertically and horizontally. It is a type of infectious disease dominated by the malignant proliferation of hematopoietic cells, including lymphocytic leukemia, erythroblastic leukemia, myeloblastic leukemia, and myeloid leukemia. As there is no effective vaccine or anti-ALV drug for ALV, detection and purification are currently the main measures to control ALV. In order to effectively purify ALV, it is essential to develop a highly sensitive, highly spec...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/702C12Q1/6804C12Q2531/113C12Q2563/107C12Q2545/113
Inventor 叶建强谢泉张建军邵红霞秦爱建钱琨高巍万志敏
Owner YANGZHOU UNIV
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