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

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

Inactive Publication Date: 2013-08-21
孙爱华 +1
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  • 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: The example establishes the rTpN15-17-47-ELISA for detecting syphilis serum antibodies by using genetic engineering technology, first obtaining tpN15, tpN17 and tpN47 gene clones, and then using the same two flexible peptide sequences to connect the outer membrane respectively Protein tpN15 and tpN17 genes, tpN17 and tpN47 genes, and construct three treponema pallidum outer membrane protein antigens containing outer membrane proteins TpN15, TpN17 and TpN47 at the same time.

[0030] The present invention adopts genetic engineering technology to construct artificial fusion gene tpN15-17-47 and its prokaryotic expression system E.coliBL21DE3 pET42a-tpN15-17-47 , the purified recombinant expression product of the expression system is coated with rTpN15-17-47 antigen, and an rTpN15-17-47-ELISA for detecting syphilis serum antibodies is established.

[0031] Gene selection basis: It has been reported in the literature that antibodies to Treponema pallidum TpN30, TpN...

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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 constructing a fusion gene with connecting primers and performing prokaryotic expression, purifying the recombinant expression product rTpN15-17-47 as a coating antigen, and establishing a method for rTpN15-17-47-ELISA to detect serum antibodies to syphilis. Background technique [0002] Syphilis caused by Treponema pallidum infection is one of the most important human sexually transmitted diseases (STD). The incubation period of syphilis is long, the progress of the disease is relatively slow, and the positive rate of Treponema pallidum isolated from the lesion is low, so 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 Hemaggluti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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