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Compositions and methods for detection of drug resistant mycobacterium tuberculosis

A technology for detecting Mycobacterium tuberculosis and samples, which is applied in the field of PCR detection using hydrolysis probes, and can solve problems such as inability to detect

Active Publication Date: 2017-08-01
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many commercially available nucleic acid tests for MTB resistance have very fast turnaround times but cannot detect small percentages of mutants in mixed infections containing both wild-type and mutants

Method used

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  • Compositions and methods for detection of drug resistant mycobacterium tuberculosis
  • Compositions and methods for detection of drug resistant mycobacterium tuberculosis
  • Compositions and methods for detection of drug resistant mycobacterium tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0104] Detection of specific SNPs using exemplary rpoB, inhA and katG probes

[0105] Exemplary probes were examined for their ability to detect: rbo 17 SNPs in the B gene that confer MTB resistance to rifampicin, including: rpo B 531L, rpo B 531W, rpo B 526L, rpo B 526Y, rpo B 526D, rpo B 526N, rpo B 513L, rpo B 513K, rpo B 513P, rpo B 522L, rpo B 522Q, rpo B 522W, rpo B 516V, rpo B 516Y, rpo B533P, rpo B 511P and rpo B 526R; inh The 3 SNPs in the A gene that confer MTB resistance to isoniazid include: inh A-15T, inh A-8A and inh A-8C; and kat Four SNPs in the G gene that also confer MTB resistance to isoniazid include: kat G315I, kat G 315N, kat G 315T and kat G 315T2.

[0106] Various plasmids containing the respective nucleic acids, each representing one, two or three SNPs, were used. In this paper, in each well, containing buffer, ZO5 DNA polymerase, nucleoside triphosphate, capable of amplifying rpoB, inhA and ka...

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Abstract

Methods for the rapid detection of the presence or absence of Mycobacterium tuberculosis (MTB) resistant to rifampicin (MTB-RIF) and / or MTB resistant to isoniazid (MTB-INH) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers, probes targeting the genes for rpoB, inhA, and katG, along with kits are provided that are designed for the detection of MTB-RIF and / or MTB-INH.

Description

[0001] field of invention [0002] The present invention relates to the field of viral diagnostics, and more particularly, to methods for detecting drug-resistant Mycobacterium tuberculosis ( Mycobacterium tuberculosis ) PCR detection method using hydrolysis probe. [0003] Background of the invention [0004] Tuberculosis (TB) is a bacterial disease most commonly found in the lungs caused by various strains of mycobacteria, such as Mycobacterium tuberculosis (MTB). TB is spread from person to person through the air when an individual with lung or throat TB coughs, sneezes, or spits, propelling MTB into the air. It is estimated that one-third of the world's population is infected with MTB, and that 9 million people develop TB each year. TB has been a major cause of human infectious disease, and drug-resistant strains of MTB are increasing, especially in developing countries. [0005] Two common first-line drugs used to treat MTB include isoniazid (INH) and rifampicin (RIF), ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/689C12Q2600/156C12Q2600/16C12Q2600/158
Inventor J.A.约翰逊R.梅塔A.袁
Owner F HOFFMANN LA ROCHE & CO AG
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