Method for detecting rifampin drug resistance of mycobacterium tuberculosis by using pyrosequencing technology

A technology of Mycobacterium tuberculosis and pyrosequencing, which is applied in the directions of microorganism-based methods, microbial determination/inspection, biochemical equipment and methods, etc. Many times, etc., to achieve the effect of shortening the detection time, reducing the number of reactions and the amount of detection reagents

Active Publication Date: 2013-04-17
SHANGHAI PULMONARY HOSPITAL
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the existing conventional methods for detecting drug resistance of Mycobacterium tuberculosis, it is unavoidable to isolate and cultivate Mycobacterium tuberculosis strains in advance from pat

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
  • Method for detecting rifampin drug resistance of mycobacterium tuberculosis by using pyrosequencing technology
  • Method for detecting rifampin drug resistance of mycobacterium tuberculosis by using pyrosequencing technology
  • Method for detecting rifampin drug resistance of mycobacterium tuberculosis by using pyrosequencing technology

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0026] Since the mutations of the rpoB gene are mainly concentrated in the RRDR region, and there are many mutation sites in the RRDR region, the pyrosequencing detection method of the present invention is mainly aimed at the RRDR region of the rpoB resistance gene of rifampin. The present invention provides a method for detecting the resistance of Mycobacterium tuberculosis to rifampicin. The rpoB gene is first amplified by PCR, and then the PCR amplified product is subjected to pyrosequencing using SQA mode to determine whether the rpoB gene has mutations Site, where the nucleotide loading sequence is AGCT.

[0027] Pyrosequencing technology

[0028] Pyrosequencing (Pyrosequencing) technology is a brand-new DNA sequence analysis technology that can quickly and accurately determine a short target fragment. This technique does not require electrophoresis, and DNA fragments do not require fluorescent labeling, and the operation is extremely simple. Because the pyrosequencing tech...

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 method for detecting rifampin drug resistance of mycobacterium tuberculosis by using a pyrosequencing technology. The method comprises the following steps: performing a PCR amplification for a rpoB gene, performing the pyrosequencing of the PCR amplification products, and determining whether or not the rpoB gene has mutation points, wherein the pyrosequencing uses a SQA mode, and a nucleotide sampling sequence is AGCT. The method for detecting the rifampin drug resistance of the mycobacterium tuberculosis by using the pyrosequencing technology can directly detect the drug resistance from phlegm samples of patients, do not need to isolate and culture M. tuberculosis strains, and thus, compared with present detection methods, can shorten detection time, and reduce reaction times and amounts of detection reagents required by the pyrosequencing.

Description

Technical field [0001] The present invention involves a method of detecting the resistance of Tuberculosis, and especially involves a primer and detection method that uses a scorched phosphate sequencing method to directly detect tuberculosis. Background technique [0002] Liping is the main first-line drug in anti-tuberculosis combined with chemotherapy. Lifepipin resistance is a symbol of MDR-TB.Lifepipin binds to the transcription and extension of RNA through the combination of β sub -units of RNA polymerase.β -sub -sub -unit of RNA polymerase encoded RNA polymers.More than 90%of the RPOB genes (RRDR) of the Rifampin Resistance RETERMING REGION (RRDR) of RPOB genes of RPOB genes in RPOB genes of more than 90%or more have genetic mutations in RRDR.The mutation type includes a single nucleotide, lack and insertion mutation.Through a large number of studies, there are 87 different types of dot mutations or short nucleotide insertions and lack of clinical separation strains of Lif...

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): C12Q1/68C12R1/32
Inventor 郑瑞娟胡忠义朱长太周燕王洁秦莲花
Owner SHANGHAI PULMONARY HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products