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Chlamydia trachomatis recombinant protein and preparation method thereof

A technology for Chlamydia trachomatis and protein, which is applied in the fields of vaccine development, genetic engineering technology and diagnostic reagents, can solve problems such as reducing detection specificity, and achieve the effect of high sensitivity and strong specificity

Inactive Publication Date: 2013-02-13
英诺特(唐山)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention provides a recombinant antigen produced by a genetic engineering method, and detection by using the recombinant antigen can avoid problems such as low detection specificity caused by false positives due to low purity or poor specificity of the protein antigen, and provides a new method for the detection of Chlamydia trachomatis infection A more specific and accurate detection raw material and detection method

Method used

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  • Chlamydia trachomatis recombinant protein and preparation method thereof
  • Chlamydia trachomatis recombinant protein and preparation method thereof
  • Chlamydia trachomatis recombinant protein and preparation method thereof

Examples

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Embodiment 1

[0024] The preparation of embodiment 1 Chlamydia trachomatis recombinant protein

[0025] 1.1 Screening of Chlamydia trachomatis protein antigen epitope and target gene cloning

[0026] Obtain the full-length sequence of the Ct143 gene of D standard strain (GenBank: NC 000117.1) from GenBank (http: / / www.ncbi.nlm.nih.gov / Entrez), design a pair of specific primers with this DNA sequence as a template, and Add NdeI and XhoI restriction sites and protective bases in the upstream and downstream respectively, and the designed specific primer sequence is as follows: F1: ATCG CATATG ATGAAGAAACCAGTATTTACAGGGGG 41.67%Tm=64.64R1: GATC CTCGAG TTAATCTGCCTCCTTATAAGAAG42.42%Tm=63.49

[0027] Using the genome of Chlamydia trachomatis D standard strain D / UW.3 / CX (purchased from China Institute for the Control of Pharmaceutical and Biological Products) as a template, PCR amplification was performed with primers F1 and R1, and the Ct143 fragment was amplified;

[0028] The 50μl PCR amplif...

Embodiment 2

[0049] Example 2 Preparation and Performance Test of Chlamydia trachomatis IgM Antibody Colloidal Gold Detection Kit

[0050] 2.1 Preparation of Chlamydia trachomatis IgM Antibody Antibody Colloidal Gold Detection Kit

[0051] The recombinant Ct protein prepared above was used as the labeled antigen of the kit, and the CT-IgM antibody was detected by the colloidal gold method. The kit was developed and used as follows:

[0052] (1) Principle of the kit

[0053] According to the principle of immune capture method, the invention uses mouse anti-human IgM (μ chain) monoclonal antibody and rabbit anti-chlamydia trachomatis antibody to coat nitrocellulose membrane, and colloidal gold marks recombinant chlamydia trachomatis (Ct) antigen as a tracer. When using, add the serum to be tested. If the sample contains CT-IgM antibody, it can combine with colloidal gold-labeled Ct antigen to form a complex. The complex is captured at the coated mouse anti-human IgM antibody and reacts wit...

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Abstract

The invention provides a chlamydia trachomatis recombinant protein and a preparation method thereof, relating to the field of gene engineering technology and diagnostic reagent and vaccine development. The invention relates to a chlamydia trachomatis fusion protein which can be applied to the clinical diagnosis of chlamydia trachomatis infection; and a nucleotide sequence coding the protein comprises plasmids of the nucleotide sequence and pronucleus host cells. The invention also relates to a method for preparing the protein and application thereof. The recombinant protein has high specificity and strong immunogenicity, and can improve the sensitivity and specificity of a detection reagent; and compared with the same kind of kits on the market, the recombinant protein has the advantages of strong specificity, high sensitivity and the like, and perfectly meets the needs in the clinical diagnosis of human chlamydia trachomatis infection.

Description

technical field [0001] The invention relates to the fields of genetic engineering technology, diagnostic reagents and vaccine development, in particular to a Chlamydia trachomatis recombinant protein and a preparation method thereof. Background technique [0002] Chlamydia trachomatis is an intracellular parasite whose infectious form is a protoplasma. The protoplasma infects the host cell, transforms into a metabolically active reticular body, proliferates through binary division, and finally produces the protoplasma, which is released from the cell to infect new cells. Urogenital tract infection caused by Chlamydia trachomatis is the most common type of sexually transmitted disease (sextransmitted disease, STD), and the infection rate is high during the sexually active period. The transmission of Chlamydia trachomatis is affected by many factors, among which age, gender, race, contraceptive measures, multiple sexual partners, previous history of Chlamydia trachomatis infe...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/295C12N15/70C12N1/21G01N33/68G01N33/569G01N33/558C12R1/01C12R1/19
Inventor 华艳陈廷友张晓刚
Owner 英诺特(唐山)生物技术有限公司
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