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A kind of recombinant mycoplasma pneumoniae protein and its application

A mycoplasma pneumoniae and protein technology, which is applied in the fields of genetic engineering, vaccine development and diagnostic reagents, can solve the problems of high requirements for mycoplasma culture, low pathogen content, low positive rate, etc., and meets the needs of clinical diagnosis, simple operation and high sensitivity Effect

Active Publication Date: 2016-08-17
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

Although isolation and culture are the most reliable basis for diagnosis, there are few pathogens in clinical specimens, more bacteria in the respiratory tract, long time for isolation and culture, low positive rate, and high requirements for mycoplasma culture, which are difficult for general laboratories to carry out. shortcoming
Therefore, it cannot be used as a rapid clinical diagnosis method

Method used

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  • A kind of recombinant mycoplasma pneumoniae protein and its application
  • A kind of recombinant mycoplasma pneumoniae protein and its application
  • A kind of recombinant mycoplasma pneumoniae protein and its application

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0041] The preparation of embodiment 1 mycoplasma pneumoniae protein

[0042] 1.1 Mycoplasma pneumoniae protein epitope screening and target gene cloning

[0043] The strong epitope of the Mycoplasma pneumoniae protein is screened out by computer analysis of the entire amino acid sequence of Mycoplasma pneumoniae, and the recombinant protein contains p116 from the N-terminal to the C-terminus of the MP129-B7 genome from the 11th to the 11th position of the MP129-B7 genome. 289 nucleotides, and 421 amino acids from 339th to 421st nucleotides. Wherein the DNA sequence of the above recombinant protein is shown in SEQ ID No.2.

[0044] According to the cDNA sequence of the target peptide in the genome of Mycoplasma pneumoniae MP129-B7 and the restriction site on the plasmid, two pairs of PCR primers (P1, P2) and (P3, P4) were designed. p1 and p2 are used to amplify the 11th to 304th nucleotides in p116, p3 and p4 are used to amplify the 389th to 465th nucleotides of p116; primer...

Embodiment 2

[0067] Preparation and Performance Test of Example 2 Mycoplasma pneumoniae Antibody Colloidal Gold Detection Kit

[0068] 2.1 Preparation of Mycoplasma pneumoniae Antibody Colloidal Gold Detection Kit

[0069] The recombinant rMP-p116 protein prepared above was used as the labeled antigen of the kit, and the MP antibody was detected by the colloidal gold method. The kit was developed and used as follows:

[0070] (1) The principle of the kit According to the principle of the double-antigen sandwich method, the present invention uses the rMP-p116 recombinant antigen to coat the nitrocellulose membrane, and the colloidal gold-labeled genetically engineered recombinant Mycoplasma pneumoniae (rMP-p116) antigen is used as a tracer. When using, add the serum to be tested, if the sample contains anti-MP specific antibody, it can combine with the rMP-p116 recombinant antigen on the membrane surface to form a complex, and the complex will combine with colloidal gold-labeled rMP-p116 a...

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Abstract

The invention relates to recombination mycoplasma pneumoniae protein and an application thereof in preparation of a detection kit. A diagnostic kit prepared from the recombination mycoplasma pneumoniae protein provided by the invention has the advantages of strong specificity, high sensitivity, simple and convenient operation and the like and meets the requirements of clinical diagnosis of mycoplasma pneumoniae infection very well.

Description

technical field [0001] The invention relates to the fields of genetic engineering, vaccine development, diagnostic reagents and the like, in particular to a mycoplasma pneumoniae protein and its application. Background technique [0002] Mycoplasma pneumoniae (Mycoplasma pneumoniae, MP) is one of the common pathogens that cause upper respiratory tract infection, tracheobronchitis and atypical pneumonia. The clinical manifestations of patients with community-acquired pneumonia caused by Mycoplasma pneumoniae infection are not typical. After an incubation period of 10-20 days, some non-specific symptoms such as headache and fever may appear, often accompanied by weakness and dry cough. It is difficult to make a differential diagnosis based on clinical symptoms. Often neglected and lead to serious complications. Since Mycoplasma pneumoniae has no cell membrane, antibacterial drugs acting on the cell membrane have no killing effect on it, and its clinical manifestations are not...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/70C12N1/21C07K14/30G01N33/68G01N33/569C12R1/19
Inventor 张秀杰张晓刚华艳
Owner 英诺特(唐山)生物技术有限公司
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