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Indirect ELISA kit for detecting porcine transfusion transmitted virus 2 (TTV2) antibody

A blood transfusion-transmitted virus and kit technology, which is applied in the field of indirect ELISA kits, can solve the problem of low conservation and achieve the effect of a simple detection method

Inactive Publication Date: 2013-08-07
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

In addition, the mutation patterns are also different among ORFs: ORF1 probably encodes the viral capsid protein (Cap) and replication-associated protein (Rep), which is relatively conserved and has a relatively high rate of change at the third position of the codon, although it has been reported that Some conserved regions of ORF2 and ORF3, but relatively speaking, the conservation of the two is relatively low

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  • Indirect ELISA kit for detecting porcine transfusion transmitted virus 2 (TTV2) antibody
  • Indirect ELISA kit for detecting porcine transfusion transmitted virus 2 (TTV2) antibody
  • Indirect ELISA kit for detecting porcine transfusion transmitted virus 2 (TTV2) antibody

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Embodiment Construction

[0045] Preparation of coating antigen:

[0046] 1.1 Design and synthesis of specific primers: According to the porcine TTV2 gene sequence included in GenBank (GenBank accession number: EF514716), the ORF1 gene encoding the porcine TTV2 open reading frame was selected, and primers containing restriction sites were designed using Primer Premier 5.0 software and analyzed. Submit to Shanghai Yingjun Company for synthesis.

[0047] SFORF1-:CCG CTCGAG GACTTAACGGAACCGTGGCTAGAAG ( xho I)

[0048] SRORF1:CCC AAGCTT TGTTTTTCATCCTCTTTACCACCCGCTGGA( Hind III)

[0049] 1.2 Construction of prokaryotic expression vector: Utilize the above-mentioned specific primers to amplify gene fragments from TTV2 positive disease materials to obtain 1227bp amplified fragments, and then use the restriction endonucleases on the respective primers to amplify the PCR amplification products and The pcoldI prokaryotic expression vector (purchased from Takara) was digested, and the digested PCR amplif...

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Abstract

The invention relates to an indirect ELISA kit for detecting a porcine transfusion transmitted virus 2 (TTV2) antibody and belongs to the field of biotechnology. The invention comprises antigen recombinant protein preparation, indirect ELISA establishing, and use of determination standard and clinical serological test. Through pcoldI prokaryotic expression vectors, a gene engineering bacterium pcoldI-ORF1 for expression of a porcine TTV2ORF1 truncated protein is constructed and the expressed antigen recombinant protein is purified and is used as an antigen so that an indirect ELISA detection method is established. The indirect ELISA detection method is used for detecting a TTV2 antibody level of porcine serum, has good repeatability and high singularity, can be used for porcine TTV2 serology investigation and is a fast and simple serological test method for prevention, treatment and prevalence state control of porcine transfusion transmitted diseases. The indirect ELISA detection method utilizes the porcine TTV2ORF1 recombinant protein to detect the porcine TTV2 antibody first in China.

Description

technical field [0001] The invention relates to an indirect ELISA kit for detecting antibodies to porcine transfusion-transmitted virus type 2 (namely porcine TTV2), belonging to the field of veterinary biotechnology. Background technique [0002] TTV (Torque Teno virus), also known as transfusion-transmitted virus, belongs to the family Anelloviridae and the genus Iotatorquevirus. It is a non-enveloped single-stranded circular negative-strand DNA virus. It was first discovered by Japanese scholar Nishizawa et al. in 1997. Since the virus was isolated from a hepatitis patient, it was suspected that it was related to hepatitis, which attracted great attention. Subsequently, various countries conducted epidemiological investigations on the TTV infection in different populations in their own countries, and found that the positive rate of TTV DNA in the population is generally above 10%. It has been confirmed that, in addition to humans can be infected with TTV, in primates (ch...

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Application Information

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IPC IPC(8): G01N33/569
Inventor 何孔旺王小敏张文文倪艳秀温立斌俞正玉张雪寒郭容利吕立新李彬周俊明茅爱华叶青汪伟周萍沈江萍
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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