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Fluorescence quantitative detection method of Ureaplasma urealyticum and Ureaplasma parvum

A technology for fluorescent quantitative detection and Ureaplasma urealyticum, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, fluorescence/phosphorescence, etc. It can solve the problems of difficult to accurately judge the results, cumbersome operation, and prone to cross-reactions , to achieve the effect of less sample demand, high detection efficiency and good specificity

Active Publication Date: 2015-10-14
上海国兴医疗器械有限公司
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Problems solved by technology

[0003] There are differences in the distribution and pathogenicity of the two biotypes of Ureaplasma in the population. The commonly used Ureaplasma serological typing method is cumbersome to operate, prone to cross-reactions, and the results are difficult to judge accurately

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  • Fluorescence quantitative detection method of Ureaplasma urealyticum and Ureaplasma parvum
  • Fluorescence quantitative detection method of Ureaplasma urealyticum and Ureaplasma parvum
  • Fluorescence quantitative detection method of Ureaplasma urealyticum and Ureaplasma parvum

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Embodiment Construction

[0027] Below in conjunction with specific embodiment, further illustrate the present invention. These examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples is usually according to conventional conditions, such as molecular cloning such as Sambrook: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's instructions suggested conditions.

[0028] Example Fluorescence quantitative detection of Ureaplasma urealyticum and Ureaplasma micronica

[0029] Step 1, design and synthesis of primers and probes

[0030] The sequences of the primers and probes are shown in the sequence table, and the positions of the primers and probes in the corresponding nucleotide sequence table and the lengths of the amplified products are shown in Table 1.

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Abstract

The invention relates to a fluorescence quantitative detection method of ureaplasma urealyticum and microureaplasma. The method comprises the following steps of: processing a sample and extracting DNA (Deoxyribonucleic Acid), and specifically comprises the following steps of: using oligonucleotide sequence complemented with ureaplasma urealyticum and microureaplasma topoisomerase IV DNA as a forward primer, a reverse primer and a fluorescently-labeled probe in PCR reaction to respectively add in two different PCR amplification tubes, simultaneously amplifying DNA target sequences of ureaplasma urealyticum and microureaplasma in the above two amplification tubes respectively by a same amplification condition; judging whether the sample to be detected containing ureaplasma urealyticum and microureaplasma or not through a standard curve according to change of different fluorescence signals in a detection reaction system, and determining the corresponding content of ureaplasma urealyticum and microureaplasma. As for identification and quantitative analysis of reaplasma urealyticum and microureaplasma, the method provided by the invention has characteristics of speediness, sensitivity, high specificity and low cost.

Description

technical field [0001] The invention relates to a fluorescent quantitative detection method, in particular to a fluorescent quantitative detection method for Ureaplasma urealyticum and Ureaplasma micron. Background technique [0002] Ureaplasma (Ureaplasma) is one of 13 kinds of mycoplasma isolated from the human body that can reproduce by itself. Ureaplasma is between viruses and bacteria. It is the smallest prokaryotic microorganism that can reproduce on artificial medium. It has no cell wall and can replicate itself. It is club-shaped, with a size of 125-250nm and a molecular weight of 4.5×10 8 , highly pleomorphic. Ureaplasma is divided into two biological groups and 14 serotypes according to molecular biological characteristics. The two biological groups include Ureaplasma parvum (Up) and Ureaplasma urealyticum (Uu), and 14 serotypes Among them, serotypes 1, 3, 6, and 14 with smaller genomes belong to U. minutum, accounting for 90% to 92% of Ureaplasma infections; ser...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/06G01N21/64
Inventor 赵缜刘璐平亚芬
Owner 上海国兴医疗器械有限公司
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