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Preparation method of rTpN15-17-47-ELISA for detecting syphilis serum antibody

A serum antibody, tpn15 technology, applied in the field of serological screening or diagnosis of syphilis, can solve the problem of low sensitivity and specificity of a single recombinant antigen

Inactive Publication Date: 2011-09-14
孙爱华 +1
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  • Description
  • Claims
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Problems solved by technology

At present, there have been many research reports on serum antibody detection methods established with a single rTpNs antigen, but the sensitivity and specificity of a single recombinant antigen are low

Method used

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  • Preparation method of rTpN15-17-47-ELISA for detecting syphilis serum antibody
  • Preparation method of rTpN15-17-47-ELISA for detecting syphilis serum antibody

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Embodiment

[0029] Example: Example: The establishment of rTpN15-17-47-ELISA for the detection of syphilis serum antibodies uses genetic engineering technology. Firstly, tpN15, tpN17 and tpN47 gene clones are obtained respectively, and then the same two flexible peptide sequences are used to connect the outer membrane. Proteins tpN15 and tpN17 genes, tpN17 and tpN47 genes, construct three outer membrane protein antigens containing TpN15, TpN17 and TpN47 simultaneously.

[0030] The present invention uses genetic engineering technology to construct artificial fusion gene tpN15-17-47 and its prokaryotic expression system E.coli BL21DE3 pET42a-tpN15-17-47 The purified recombinant expression product rTpN15-17-47 of the expression system was coated with antigen to establish rTpN15-17-47-ELISA for detecting syphilis serum antibodies.

[0031] Gene selection basis: It has been reported in the literature that TpN30, TpN33 and TpN37 antibodies are present in the serum of patients with early syphilis, bu...

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Abstract

The invention relates to a syphilis serological screening or diagnosis method. The purpose is that the method has the characteristics of low price, high speed, sensitivity and specificity. The technical scheme provided by the invention is that: a preparation method of rTpN15-17-47-ELISA for detecting syphilis serum antibodies comprises the following steps in turn: constructing an artificial fusion gene tpN15-17-47 of tpN15, tpN17 and tpN47 and a prokaryotic expression system E.coliBL21DE3Pet42a-tpN15-17-47 by gene technology, purifying the recombinant expression product rTpN15-17-47 as a coating antigen, establishing rTpN15-17-47-ELISA for detecting syphilis serum antibodies. The fusion gene tpN15-17-47 has a nucleotide sequence and an amino acid sequence as shown in sequence table 5.

Description

Technical field [0001] The present invention relates to a rapid, sensitive and specific serological screening or diagnosis method for syphilis. Specifically, it uses connecting primers to construct a fusion gene and perform prokaryotic expression, and purify the recombinant expression product rTpN15-17-47 as a coating antigen. RTpN15-17-47-ELISA for detecting syphilis serum antibodies. Background technique [0002] Syphilis (syphilis) caused by Treponema pallidum infection is one of the most important human sexual transmission diseases (STD). The incubation period of syphilis is long, the disease progression is relatively slow, and the positive rate of Treponema pallidum isolated from the lesion is low. Therefore, the serological test for detecting Treponema pallidum specific antibodies has become a clinical laboratory diagnostic method for syphilis. For example, TRUST (Syphilis toluidine red unheated serum test) and TPHA (Treponema pallidum antibody hemagglutination test) are c...

Claims

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Application Information

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IPC IPC(8): G01N33/571G01N33/531C12N15/31C12N15/62C12N15/70C12N1/21C07K19/00
Inventor 孙爱华严杰
Owner 孙爱华
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