Method for quickly detecting gonorrhea Neisseria by utilizing loop-mediated isothermal amplification technology

A Neisseria gonorrhoeae, loop-mediated isotherm technique for application in the field of biomedical diagnostics

Inactive Publication Date: 2010-09-15
孙星江
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, it has not been reported at home and abroad to apply it to the detection of Neisseria gonorrhoeae

Method used

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  • Method for quickly detecting gonorrhea Neisseria by utilizing loop-mediated isothermal amplification technology
  • Method for quickly detecting gonorrhea Neisseria by utilizing loop-mediated isothermal amplification technology
  • Method for quickly detecting gonorrhea Neisseria by utilizing loop-mediated isothermal amplification technology

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Experimental program
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Embodiment approach

[0062] Use the QIAamp DNA Mini Kit (QIAGEN Company) to extract high-purity DNA from the patient's urethral secretion or urine centrifugal sedimentation samples, and the specific operation is completed according to its instructions. (This step can also be completed using similar DNA extraction kits from other biological companies). 50 μl of DNA samples from urethral secretions of gonorrhea patients were recovered by using the above kit, which was labeled as sample 2; and 50 μl of DNA samples from healthy human urethral secretions were labeled as sample 1. Take 2 μl from each of sample 1 and sample 2 into a new small PCR tube, and then add 50 μl of reaction solution (the components are as follows). Close the cap of the PCR tube. Place the tube in a constant temperature metal bath at 65°C for 1 hour.

[0063] Then take out the tube and observe the color of the liquid in the tube with naked eyes. If the liquid in the tube turns green, it indicates that the specific nucleic acid...

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Abstract

The invention belongs to biomedical diagnosis, and discloses a method for detecting gonorrhea Neisseria by utilizing a loop-mediated isothermal amplification technology and a sequence of a primer used by the method. By combining the loop-mediated isothermal amplification technology with the fluorochrome development, the method can realize quick isothermal nucleic acid amplification of the specific DNA sequence of gonorrhea Neisseria and result detection on the premise of not using a PCR amplifier, an electrophoresis apparatus or an ultraviolet gel imaging system. The method is simple to operate, and can be used for detecting the gonorrhea Neisseria in urethral secretions of a clinic outpatient or serves as a method for self-check of patients.

Description

technical field [0001] The invention belongs to the field of biomedical diagnosis, and in particular relates to a method for rapidly and conveniently detecting Neisseria gonorrhoeae using loop-mediated isothermal amplification (LAMP) technology. It can be used for clinical diagnosis and personal early screening. Background technique [0002] Neisseria gonorrhoeae (Neisseria gonorrhoeae) is a common pathogen of genitourinary transmission diseases. The traditional clinical detection method is to use the bacterial culture method, but this method is difficult to operate, takes a long time, the conditions for specimen transportation are extremely strict, and it also has the disadvantage of low detection rate, which easily leads to missed diagnosis and misdiagnosis. In addition, this method also needs to obtain samples invasively, which causes great pain to patients. Conventional PCR detection methods can avoid this problem, and there are some commercial kits that use nucleic ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/36
Inventor 孙星江
Owner 孙星江
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