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PCR (Polymerase Chain Reaction) kit for detecting ureaplasma urealyticum biogroup

A Ureaplasma urealyticum and kit technology, which is applied in the field of kits for detecting Ureaplasma urealyticum biota, can solve the problems of high cost, time-consuming, poor specificity of results, and susceptibility to contamination, so as to reduce the chance of contamination, The effect of improving specificity and increasing the degree of automation

Inactive Publication Date: 2013-10-23
SHANGHAI PULMONARY HOSPITAL
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Problems solved by technology

This method is not only costly and time-consuming (usually several days to a week), but also immune sera are prone to cross-reactivity (type 5 and type 2, type 14 and type 3, type 8 and type 13), resulting in poor specificity of the results
Although conventional PCR technology can complete the rapid detection of Ureaplasma urealyticum biota, it is easily contaminated, false positives occur, and quantitative detection cannot be completed
[0008] In view of the lack of a better method for rapid grouping of U. urealyticum in clinical practice, it is urgent to develop a new method for the detection of U. urealyticum biota

Method used

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Embodiment Construction

[0031] The invention provides a primer, a kit, and a detection method for real-time quantitative PCR detection of Ureaplasma urealyticum organisms, and a fluorescent reporter group 6-carboxyfluorescein (6-carboxyfluorescein (6- carboxyfluorescein, FAM), etc., with a quenching fluorescent group 6-carboxytetramethyldanoamine (TAMRA) added to the 3′ end. When the probe structure is intact, the quencher group at the 3' end inhibits the fluorescence emission of the fluorophore at the 5' end.

[0032] As the PCR reaction proceeds, due to the 5'→3' exonuclease activity of Taq enzyme, when the synthesized new strand moves to the probe binding position, Taq enzyme cuts off the probe, and the integrity of the probe is destroyed. The energy transfer structure is also destroyed, and the fluorescent signal of the FAM fluorescent reporter group at the 5' end is released. Every time the template is replicated, a probe is cut off, accompanied by the release of a fluorescent signal. As the p...

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Abstract

The invention relates to a PCR (Polymerase Chain Reaction) detection kit and particularly relates to a kit used for detecting ureaplasma urealyticum biogroup and a detecting method. The invention further provides two groups of primers SEQIDNo.1-4. Compared with the common PCR technology, the invention has the advantages that the detection specificity and the detection sensitivity are greatly improved, the pollution opportunity is reduced, and the automation degree is improved as well. The kit used for detecting ureaplasma urealyticum (UU) biogroup not only achieves rapid diagnosis (within 3 hours), but also performs quantitative analysis on the UU biogroup.

Description

technical field [0001] The invention relates to a PCR detection kit, in particular to a kit for detecting Ureaplasma urealyticum organisms and a detection method. Background technique [0002] Ureaplasma urealyticum ( Ureaplasma urealyticum UU), also known as Ureaplasma urealyticum, can use its own urease to decompose urea to provide energy for metabolism. It is one of the common parasites in the human urogenital tract and is closely related to human urogenital infection. Has now been listed as the pathogen of sexually transmitted diseases. The pathogenic substances of Ureaplasma urealyticum mainly include urease, IgA protease and phospholipase. [0003] The research on the relationship between Ureaplasma urealyticum and diseases started from non-gonococcal urethritis. Therefore, the harm of Ureaplasma urealyticum to human health cannot be ignored, and it is necessary to pay attention to it. [0004] In the past, Ureaplasma urealyticum infection was mainly diagnosed by e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06C12N15/11G01N21/64
Inventor 朱长太郑瑞娟胡忠义刘忠华
Owner SHANGHAI PULMONARY HOSPITAL
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