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Diagnostic marker for ureaplasma urealyticum infection and preparation method and application of detection kit corresponding to diagnostic marker

Ureaplasma urealyticum and detection kit technology, applied in the field of detection kit preparation, can solve the problems of loss of cell components, false negatives, many influencing factors, etc., and achieve the effects of high repeatability of results, simple operation, and objective judgment of results

Pending Publication Date: 2021-06-11
THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the domestic indirect method for detecting mycoplasma antibodies should be a qualitative reagent, but because mycoplasma exists widely in the population, and normal people also have low-titer antibody levels, it is not possible to rely solely on qualitative antibody detection for diagnosis; in addition, monoclonal antibody immunofluorescence detection There are many influencing factors of urealyticum, such as not taking cell components during smear, and loss of cell components during fixation can also cause false negative results

Method used

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  • Diagnostic marker for ureaplasma urealyticum infection and preparation method and application of detection kit corresponding to diagnostic marker
  • Diagnostic marker for ureaplasma urealyticum infection and preparation method and application of detection kit corresponding to diagnostic marker
  • Diagnostic marker for ureaplasma urealyticum infection and preparation method and application of detection kit corresponding to diagnostic marker

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Experimental program
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Effect test

Embodiment 1

[0031] The preparation flow chart of this embodiment is as follows figure 2 As shown, the figure includes the following steps:

[0032] 1. Preparation of UUR10_0579 protein from Ureaplasma urealyticum

[0033]According to the characteristics of strong antigenicity, good surface and small molecular weight of UUR10_0579 protein of Ureaplasma urealyticum, the experimental bioinformatics software analyzed the amino acid sequence of UUR10_0579 protein, predicted and determined the epitope site on the UUR10_0579 protein, and included the UUR10_0579 protein by artificial synthesis. The polypeptide at the antigenic determinant site (as shown in sequence 1) has a purity of more than 95% after purification.

[0034] The amino acid sequence of sequence 1 is as follows:

[0035] MLLTIAFYLSKTTHYLKKNFNYFLDLLNQNKKHIELIIIDDASDYNLFKTLKPLIENTNSKIKYFYLNETQGNAYAYNLATKYAHGKYIWYLGGHTELNLDASSLLFSVLEKDYDVISFNLNDNVNQNPSLVFDSLNKEVLVGLWESISNKIIALDFIKKHQLAFYNDKWYPALFIYDLFTKFSSWRNVNVNFISNNSGEVGYNVYDLLQ...

Embodiment 2

[0047] The specific operation process of the kit in Example 1 to detect the specific antibody in the UUR10_0579 protein serum is as follows:

[0048] Specific steps for implementing the operation: (1) Adding samples: Take the serum to be tested, add serum sample dilution buffer, and mix well; (2) Incubation: add one drop of calf serum and one drop of the serum sample to be tested in the well, and set a negative / positive control Add two drops of negative control or positive control to each well, and seal the plate at 37°C for 1 hour; (3) Washing: Shake off the liquid in the wells of the microplate, inject washing solution, let it stand for 10 seconds, and dry it. And repeat 4 times; (4) Add enzyme-labeled antibody: add 1 drop of enzyme-labeled antibody to each well, and seal the plate at 37°C for 1 hour; (5) Wash the plate as before; (6) Color development: add chromogenic reagent A One drop each of developer B and one drop of chromogen B, and stand at 37°C for 20 minutes; (7) T...

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Abstract

The invention discloses a preparation method and application of a diagnostic marker for ureaplasma urealyticum infection and a detection kit corresponding to the diagnostic marker, the diagnostic marker is a polypeptide containing a UUR10_0579 protein antigenic determinant site, and the amino acid sequence of the diagnostic marker is shown as a sequence 1. According to the invention, pathogen-related specific antibodies are detected through quantitative ELISA, and the kit has the advantages of rapidness, simplicity, high throughput and high result repeatability; and secondly, the UUR10_0579 protein is an important antigen for early infection of the ureaplasma urealyticum and participates in immune response of an organism, so that the UUR10_0579 protein can be used as a specific diagnostic marker for infection of the ureaplasma urealyticum. Compared with a current clinical method for detecting the ureaplasma urealyticum, the method for detecting the ureaplasma urealyticum infection, provided by the invention, has the advantages of early stage, rapidness, sensitivity, specificity, accuracy and the like, and overcomes the defects of the current clinical detection method. The detection method is objective in result judgment and simple and convenient to operate, is suitable for large-scale screening tests, and can be used for final definite diagnosis of ureaplasma urealyticum infection.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a preparation method and application of a diagnostic standard substance for ureaplasma urealyticum infection and a corresponding detection kit. Background technique [0002] Ureaplasma urealyticum (M. urealyticum, UU), also known as Ureaplasma urealyticum, is one of the six genus Ureaplasma, and is closely related to human genitourinary tract infection. The bacterium is tiny, only one-fifth of Escherichia coli, and it is the smallest and simplest prokaryote known to have the ability to reproduce itself. UU is mainly distributed in the genitourinary tract of the human body, and is the most common pathogenic bacteria of non-gonococcal urethritis except Chlamydia trachomatis. It invades the mucosal cells of the female reproductive tract, causing vaginitis and cervicitis, and continues to ascend the infection, adsorbed on the fallopian tube mucosa, causing t...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/31G01N21/78
CPCG01N33/68G01N21/31G01N21/78
Inventor 张军华陈列松陆路黄蓉张晶晶
Owner THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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