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Ureaplasma urealyticum PCR (Polymerase Chain Reaction) detection kit and detection method thereof

Ureaplasma urealyticum and detection kit technology, applied in the field of microbial detection, can solve problems such as strengthening cross-reaction, reducing specific staining, non-standard experimental scheme, etc., achieving high sensitivity and specificity, good DNA extraction effect, Objective and reliable results

Active Publication Date: 2012-04-11
泰普生物科学(中国)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] MDI analysis method: As a method of biopsy, the method of MDI analysis of mycoplasma has been controversial in the industry. This technology has the following shortcomings: there are as many as 7 kinds of mycoplasma that can be detected in genitourinary system diseases. There are difficulties in type identification; both L-form bacteria and mycoplasma lack cell walls, and the two are difficult to distinguish under a microscope; there is no standardized experimental plan in China, and the standardization of test results needs to be improved, but this technology is economical, intuitive and fast. typical characteristics of
However, the sensitivity of immunological binding detection is not high, and the mycoplasma fixed on the NC membrane sometimes reduces the specific staining and strengthens the cross-reaction between each mycoplasma

Method used

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  • Ureaplasma urealyticum PCR (Polymerase Chain Reaction) detection kit and detection method thereof
  • Ureaplasma urealyticum PCR (Polymerase Chain Reaction) detection kit and detection method thereof
  • Ureaplasma urealyticum PCR (Polymerase Chain Reaction) detection kit and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Table 1: Kit Composition

[0051]

[0052]

[0053] The formula of UU-PCR reaction solution is shown in Table 2:

[0054] Table 2

[0055]

[0056]

[0057] Preparation of basic reagents:

[0058] 1.1 Tris: analytically pure, a product from a qualified supplier, content 99.7%, infrared qualified, pH (5% water) 10.3-10.9, moisture 0.3%, melting point 167-171°C, qualified absorption system, highest content of impurities qualified.

[0059] 1.2 NaOH: analytically pure, the product of Beijing Chemical Reagent Factory or a qualified supplier.

[0060] 1.3 TritonX100: brown oily liquid, soluble in cold water, and has strong foaming properties.

[0061] 1.4 EDTA: analytically pure, a product from qualified suppliers, white crystalline powder, soluble in water, the solution is acidic, hardly soluble in alcohol, the content is not less than 99.5%, the aqueous solution reaction is qualified, and the complexing force test is qualified , the highest impurity conten...

Embodiment 2

[0076] Embodiment 2 The use of kit of the present invention

[0077] 1. Reagent preparation (reagent preparation area)

[0078] 1. Take out the DNA extraction solution and set aside.

[0079] 2. After determining the number n of reaction tubes to be performed (number of samples + negative + strong positive quality control + weak positive quality control + 4 quantitative reference substances), take out the UU-PCR reaction solution and mix n×44μl UU-PCR Reaction solution, n×1μl DNA polymerase system was added to a centrifuge tube and oscillated to mix well. After instantaneous centrifugation, aliquot 45μl into each PCR reaction tube, cover the tube cap and transfer to the sample loading area, and store in the dark for 4 ℃ refrigerator for later use.

[0080] 3. Transfer the reference substance and quantitative reference substance to the sample processing area, and put them in a 4°C refrigerator for later use.

[0081] 2. Sample processing (sample processing area)

[0082] 1....

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Abstract

The invention relates to the field of microorganism detection, in particular to an ureaplasma urealyticum PCR (Polymerase Chain Reaction) detection kit and a detection method thereof, aims to provide the ureaplasma urealyticum PCR detection kit and the detection method thereof for detecting a specific nucleic acid sequence of ureaplasma urealyticum in a clinical sample so as to fulfill the aim ofquickly judging the existence of the ureaplasma urealyticum. In order to fulfill the aim, the technical scheme is to provide the ureaplasma urealyticum PCR detection kit, which includes DNA (Deoxyribonucleic Acid) extract, PCR reaction solution, dNTPs, tap enzyme, UNG enzyme, positive control, negative control and a quantitative reference product, wherein the PCR reaction solution includes a primer A and a fluorescent probe A; and the primer is divided into an upstream primer A and a downstream primer A. The kit has the advantages of high flexibility and specificity, stability, timeliness, convenience in operation and the like.

Description

technical field [0001] The invention relates to the field of microorganism detection, in particular to a PCR detection kit for Ureaplasma urealyticum strains and a detection method thereof. Background technique [0002] Ureaplasma urealyticum (Ureaplasmaurealyticum for short UU) is one of the common parasites in the human genitourinary tract, which can cause disease under certain circumstances. The colonization in the human body may have a secondary upward trend, that is, the mother's birth canal infects the newborn during delivery, and then decreases rapidly; it gradually increases after sexual life. In recent years, urogenital tract infection caused by Ureaplasma urealyticum has been paid more and more attention, and it is one of the pathogens causing nongonococcal urethritis. Has now been listed as the pathogen of sexually transmitted diseases. [0003] Ureaplasma urealyticum is one of the common pathogens of genitourinary tract infection, usually superficial infection,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04G01N21/64
Inventor 何华东姚雪
Owner 泰普生物科学(中国)有限公司
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