Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

DNA aptamer of mycobacterium tuberculosis standard strain H37Rv and preparation method thereof

A technology of Mycobacterium tuberculosis and aptamers, applied in the field of molecular biology, can solve problems such as low sensitivity, high requirements for experimental conditions and technical personnel, and long time-consuming tuberculosis, and achieve high specificity effects

Inactive Publication Date: 2015-09-30
SHANGHAI PULMONARY HOSPITAL
View PDF9 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its high cost, complicated operation, and high requirements for experimental conditions and technical personnel limit its popularization and application in clinical testing.
[0004] The current diagnosis of tuberculosis has problems such as time-consuming and low sensitivity. Exploring fast and simple diagnostic methods has always been the focus of research by scholars at home and abroad.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DNA aptamer of mycobacterium tuberculosis standard strain H37Rv and preparation method thereof
  • DNA aptamer of mycobacterium tuberculosis standard strain H37Rv and preparation method thereof
  • DNA aptamer of mycobacterium tuberculosis standard strain H37Rv and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0030] The present invention also provides a standard strain of Mycobacterium tuberculosis H 37 The ssDNA adapter preparation method of Rv comprises the following steps:

[0031] Step 1, construct random single-stranded oligonucleotide library: Design and synthesize a random single-stranded DNA library of 78 base pairs: 5'-GGGAGCTCAGAATAAACGCTCAA-N 35 -TTCGACATGAGGCCCGGATC-3', where N represents any one of the bases AGCT, with a capacity of 10 14 -10 15 , and further obtain a purified single-stranded ssDNA library for the standard strain H 37 SELEX technology screening of Rv aptamers;

[0032] Step 2, use the microwell plate as the separation medium and use the aptamer technology to screen H 37 Rv aptamer: H with coating buffer 37 Rv is coated in a microwell plate, and blank anti-screen holes, non-tuberculous mycobacteria and non-mycobacterium anti-screen holes are set at the same time; the ssDNA library and SELEX binding buffer are mixed and incubated with the anti-scree...

Embodiment 1

[0038] In the present embodiment, first prepare Mycobacterium tuberculosis standard strain H 37 The ssDNA aptamer of Rv, then utilizes it to carry out bacteriological detection, comprises the following steps:

[0039] step 1:

[0040] (1) Preparation of strains to be tested:

[0041] Mycobacterium tuberculosis standard strain H 37 Rv and 15 kinds of nontuberculous mycobacteria (NTM) were transferred to Michaelis 7H9 liquid medium containing 10% OADC (containing oleic acid, albumin, glucose and catalase) nutritional supplements, and cultivated to logarithmic at 37 ° C During the growth period, transfer to a 1.5ml centrifuge tube, centrifuge at 12,000 rpm for 5 minutes, wash twice with 1×PBS, bathe in 80°C water, and extinguish the fire for 30 minutes. After extinguishing the fire, transfer to the grinding tube, and adjust the turbidity to 1mg / ml after grinding.

[0042] The 8 kinds of non-mycobacteria were all scraped from the blood plate to grow well colonies, and treated ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a DNA aptamer of a mycobacterium tuberculosis standard strain H37Rv and a preparation method thereof. The ssDNA aptamer of the mycobacterium tuberculosis standard strain H37Rv provided by the invention is high in affinity and specificity, capable of detecting mycobacterium tuberculosis, nontuberculosis mycobacteria and nonmycobacteria with a high specificity in case of being used in microbiological detection, and capable of providing a beneficial basis for laboratory diagnosis on tuberculosis.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a standard strain of Mycobacterium tuberculosis H 37 The DNA aptamer of Rv and its preparation method. Background technique [0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium Tuberculosis (MTB) infection. It is a serious public health problem at home and abroad and is listed as one of the major infectious diseases in my country. Tuberculosis is one of the diseases with the highest morbidity and mortality in history. In 2013, there were about 9 million tuberculosis infected patients worldwide, and 1.5 million deaths due to tuberculosis; my country is one of the countries with a high burden of tuberculosis in the world, so an accurate and rapid method for detecting Mycobacterium tuberculosis is established for early diagnosis and treatment of tuberculosis has great significance. [0003] Currently, bacteriological detection of Mycobacterium tuberculosis...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/115C12N15/10G01N33/569
Inventor 秦莲花胡忠义杨华如斯坦木江·艾麦提
Owner SHANGHAI PULMONARY HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com