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Recombined chlamydia trachomatis protein and application thereof

A technology for Chlamydia trachomatis and protein, which is applied in the fields of vaccine development, diagnostic reagents, and genetic engineering, can solve the problems of false positives, complicated operations, and reduced detection specificity, and achieves the effects of simple operation, high sensitivity and strong specificity.

Inactive Publication Date: 2014-05-28
英诺特(唐山)生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the Chlamydia cell culture method is the most sensitive and reliable, but due to the cumbersome operation, high cost, culture time, and the impact of specimen collection, transportation and storage, and experimental techniques, it is difficult for general laboratories to carry out this method.
Due to the application of specific primers, the nucleic acid amplification method greatly enhances the sensitivity and specificity of detection, but it requires high laboratory conditions, equipment, and professional quality of personnel, and is prone to false positives due to cross-contamination of nucleic acid amplification products. positive
Therefore, it is hoped to obtain a method that can use recombinant antigens for detection, avoid the problems of lower detection specificity caused by low purity of protein antigens or false positives caused by specificity detection, and provide a higher specificity and more accurate detection method for Chlamydia trachomatis infection. detection proteins, detection methods and detection kits

Method used

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  • Recombined chlamydia trachomatis protein and application thereof
  • Recombined chlamydia trachomatis protein and application thereof
  • Recombined chlamydia trachomatis protein and application thereof

Examples

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Effect test

Embodiment 1

[0038] The preparation of embodiment 1 Chlamydia trachomatis protein

[0039] 1.1 Screening of Chlamydia trachomatis protein antigen epitope and target gene cloning

[0040] A strong antigenic epitope of the Chlamydia trachomatis protein is screened out by computer analysis of the entire amino acid sequence of Chlamydia trachomatis, and the recombinant protein contains sequentially from the N-terminal to the C-terminal of p116 in the CT129-B7 genome from the 11th to the 11th position of the N-terminal The 304th nucleotide, and the 421 amino acids from the 339th to the 465th nucleotide. Wherein the DNA sequence of the above recombinant protein is shown in SEQ ID No.2.

[0041] According to the cDNA sequence of the target peptide in the Chlamydia trachomatis CT PGP3-D genome and the restriction site on the plasmid, a pair of PCR primers (P1, P2) were designed. Primers p1 and p2 have restriction sites for BamH1 and XhoI, respectively.

[0042] The primer sequences are as follo...

Embodiment 2

[0061] Example 2 Preparation and Performance Verification of Chlamydia Trachomatis Antibody Colloidal Gold Detection Kit

[0062] 2.1 Preparation of colloidal gold detection kit for Chlamydia trachomatis antibody

[0063] The recombinant PGP3-D protein prepared above was used as the labeled antigen of the kit, and the CT antibody was determined by the colloidal gold method. The kit was developed and used as follows:

[0064] (1) Kit principle In this invention, according to the principle of immune capture method, mouse anti-human IgM (anti-μ chain) monoclonal antibody is used to coat nitrocellulose membrane, and colloidal gold is used to mark genetically engineered recombinant Chlamydia trachomatis (PGP3-D) antigen as shown trace. When using, add the serum to be tested. If the sample contains anti-CT specific antibody, it can combine with the mouse anti-human IgM (anti-μ chain) antibody recombinant antigen on the surface of the membrane to form a complex, which is combined w...

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Abstract

The invention relates to a recombined chlamydia trachomatis protein and also relates to application of the recombined chlamydia trachomatis protein in preparation of detection kits. The diagnosis kit prepared by using the recombined chlamydia trachomatis protein has advantages of strong specificity, high sensitivity, and simple operation and better meets the requirement of clinical diagnosis of chlamydia trachomatis infection.

Description

technical field [0001] The invention relates to the fields of genetic engineering, vaccine development, diagnostic reagents and the like, in particular to a chlamydia trachomatis protein and its application. Background technique [0002] According to the "Berger's System Handbook" (1984), Chlamydia trachomatis (CT) is divided into three biotypes, namely, mouse biotype (biovarmouse), trachoma biotype (biovartrachoma) and venereal disease lymphogranuloma biotype (biovar lyCThogranulomavenereum, LGV), the latter two are related to human diseases. Using indirect micro-immunofluorescence test, trachoma biotypes are divided into A, B, Ba, C, D, Da, E, F, G, H, I, Ia, J, K14 serotypes, and LGV biotypes are further divided into L1, L2, L2a, L34 serotypes. [0003] 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 divisi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/70C12N1/21C07K14/295G01N33/68G01N33/569C12R1/19
Inventor 张晓刚张秀杰华艳
Owner 英诺特(唐山)生物技术有限公司
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