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Fast detection method of ureaplasma urealyticum based on RPA

A technology of Ureaplasma urealyticum and detection method, which is applied in the field of rapid detection of Ureaplasma urealyticum based on RPA, can solve the problems of low specificity, low sensitivity, improvement space, etc., and achieves good specificity and high sensitivity , the effect of high specificity

Inactive Publication Date: 2020-08-14
台州市中心医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to provide a kind of rapid detection method to Ureaplasma urealyticum based on RPA, to solve the existing Ureaplasma urealyticum detection RPA method that proposes in the above-mentioned background technology also has some shortcomings, as low specificity, The shortcomings of low sensitivity, tedious and complicated, etc. make it difficult to quickly detect Ureaplasma urealyticum, and there is room for improvement in the process of rapid detection of Ureaplasma urealyticum

Method used

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  • Fast detection method of ureaplasma urealyticum based on RPA
  • Fast detection method of ureaplasma urealyticum based on RPA
  • Fast detection method of ureaplasma urealyticum based on RPA

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Effect test

Embodiment 1

[0056] Material preparation:

[0057] S1: Bacterial strains, pathogenic culture fluids that are confirmed to be UU positive by clinical culture methods; pathogenic culture fluids that are clinically confirmed and cultured; 30 rows of clinical female vaginal swabs;

[0058] S2: Reagent;

[0059] S3: instrument;

[0060] Design and synthesis of primers and probes:

[0061] S1: The 16sRNA of Ureaplasma urealyticum and the specific sequence of the multiband antigen gene were found in the NCBI database, and Oligo7 was used to design specific primers and probes. At the same time, the designed primers were specifically screened on NCBI, and the primers were handed over to the biological Synthesized by an engineering company, among them, the length of the primer: RPA primer is longer than the typical PCR primer, which is 30 nucleotides; the sequence of the primer: avoid the presence of unusual sequences in the primer, such as a long sequence consisting of a specific nucleoside Acid...

Embodiment 2

[0074] A kind of rapid detection method to Ureaplasma urealyticum based on RPA, comprises the following steps:

[0075] Material preparation:

[0076] S1: Bacterial strains, pathogenic culture fluids that are confirmed to be UU positive by clinical culture methods; pathogenic culture fluids that are clinically confirmed and cultured; 30 rows of clinical female vaginal swabs;

[0077] S2: Reagent;

[0078] S3: instrument;

[0079] Design and synthesis of primers and probes:

[0080] The initially obtained primer sequences are shown in Table 1, and the finally selected primer and probe sequences are shown in Table 2 below:

[0081]

[0082] Note: FAM-dT means base T modified with FAM, THF means tetrahydrofuran residue, BHQ1-dT means base modified with BHQ1, and C3 Spacer means C3-interarm:

[0083] S1: The 16sRNA of Ureaplasma urealyticum and the specific sequence of the multiband antigen gene were found in the NCBI database, and Oligo7 was used to design specific primers...

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Abstract

The invention concretely discloses a fast detection method of ureaplasma urealyticum based on RPA, and belongs to the technical field of ureaplasma urealyticum detection. The method comprises the following step of material preparation: Step 1, strains, and Step 2, reagents. The fast detection method of the ureaplasma urealyticum based on RPA has the advantages that the RPA technology is used for the fast detection of the ureaplasma urealyticum; meanwhile, high sensitivity and good specificity are realized; and the fast detection on the ureaplasma urealyticum is completed by a built RPA amplification method. Compared with a reference pathogen, only the ureaplasma urealyticum shows a typical amplification curve; during the detection of the ureaplasma urealyticum based on RPA, the lowest detection limit is 3 pg; and clinical case sample evaluation detection effects prove that the RPA method has the feasibility, and the clinical detection requirements can be met. A built ureaplasma urealyticum real-time fluorescent RPA method has the advantages of high specificity, high sensitivity, simplicity, convenience, high speed and the like, can be used for fast detection of the ureaplasma urealyticum, and provides feasibility and good study basis for the further studies.

Description

technical field [0001] The invention relates to the technical field of detection of Ureaplasma urealyticum, in particular to a rapid detection method for Ureaplasma urealyticum based on RPA. Background technique [0002] Ureaplasma urealyticum is a group of pathogenic microorganisms that mainly reside in the urogenital tract of the human body. When the body's immunity is weakened, it often causes diseases such as non-gonococcal urethritis (NUG), cervicitis, pelvic inflammatory disease, orchitis, and epididymitis. It can also cause infertility in men and women. Infection during pregnancy can lead to adverse pregnancy outcomes such as miscarriage, premature delivery, and premature rupture of membranes. In addition, such pathogens invade and destroy the epithelial cells of the urogenital tract mucosa, so it is more likely to cause secondary infections of other sexually transmitted diseases. One of the important pathogens of sexually transmitted diseases. Ureaplasma urealyticum...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12R1/35
CPCC12Q1/689C12Q1/6844C12Q2521/507C12Q2522/101C12Q2563/107
Inventor 王毅超谢姣贵杨合慧陈晓英吴波
Owner 台州市中心医院
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