Babesia gibsoni antibody latex agglutination detection kit as well as preparation method and application thereof

A technology of Babesia canis and a detection kit is applied in the field of immune detection, which can solve the problems of lack of molecular detection technology and serological detection technology that has not been widely used, and achieves simple operation, stable detection effect, and easy mastery. Effect

Active Publication Date: 2021-12-03
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Molecular and serological assays have not been widely used due to lack of commercial kits

Method used

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  • Babesia gibsoni antibody latex agglutination detection kit as well as preparation method and application thereof
  • Babesia gibsoni antibody latex agglutination detection kit as well as preparation method and application thereof
  • Babesia gibsoni antibody latex agglutination detection kit as well as preparation method and application thereof

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Embodiment

[0028] 1. Extract the RNA of Babesia canis, reverse transcribe it into cDNA, and use GAPDH as the internal reference gene. Real-time quantitative PCR detects that two genes, BgP47 and BgSA1, are highly expressed in the worm. Therefore, these two genes were selected for the establishment of subsequent detection methods.

[0029] 2. Construction of recombinant expression plasmids:

[0030] (1) Gene cloning:

[0031] Cloning of the BgSA1 gene:

[0032] The accession number of the genome sequence of Babesia canis BgSA1: AB246895.

[0033] A pair of primers for amplifying the full-length of BgSA1 gene were designed according to the full-length BgSA1 gene sequence: upstream primer SA1-F: 5'-GGCACGAGCAAATGTTATTATTGACATA-3', downstream primer SA1-R: 5'-TCAGACCTGATGCATTGCCCACA-3'.

[0034] Design a pair of expression primers based on the obtained full-length BgSA1 gene: upstream primer ExpSA1-F: 5'- GGATCC ACACAAATAACAACACCTG-3', the underlined part is BamHI, the downstream primer...

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Abstract

The invention discloses a babesia gibsoni antibody latex agglutination detection kit, the kit comprises a latex reagent which is commonly sensitized by babesia gibsoni surface antigen BgP47 recombinant protein and secreting antigen BgSA1 recombinant protein, the amino acid sequence of the surface antigen BgP47 recombinant protein is shown as SEQ ID No.2, and the amino acid sequence of the recombinant protein secreting the antigen BgSA1 is as shown in SEQ ID No. 1. The kit is used for detecting the canine babesia gibsoni antibody, and has the advantages of high specificity, high sensitivity, simplicity, convenience and rapidness in operation and the like.

Description

technical field [0001] The invention belongs to the field of immunodetection, and in particular relates to a latex agglutination detection kit for Babesia canis antibody and its preparation method and application. [0002] technical background [0003] Canine babesiosis is a parasitic disease mainly characterized by hemolysis caused by a variety of Babesia species of the genus Babesia. The clinical symptoms of dogs with babesiosis mainly include high fever, progressive anemia, splenomegaly, and unilateral or bilateral renal enlargement, which can lead to death in severe cases. Babesia is prevalent all over the world, and the main Babesia canis reported in my country are Babesia gibsoni and Babesia veschleri. Among tick-borne diseases, Babesia infection has a high proportion. [0004] The currently commonly used detection method is blood smear staining microscopy. Although this method is simple to operate and has strong specificity, its sensitivity is not high, and it depend...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N33/569G01N33/546C07K14/44
CPCG01N33/56905G01N33/54313C07K14/44G01N2333/44G01N2469/20Y02A50/30
Inventor贺兰张红艳赵俊龙崔杰战雪燕亢婷
OwnerHUAZHONG AGRI UNIV