Kit for detecting or assisting in detecting ethambutol resistance of multiple drugs resistant bacillus tuberculosis

A technology of Mycobacterium tuberculosis and ethambutol, which is applied in the detection/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of high treatment costs, high mortality, and long treatment cycles, and achieve Effective early rapid diagnosis and high diagnostic sensitivity

Inactive Publication Date: 2013-04-24
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Multidrug-resistant tuberculosis not only has a long treatment cycle, large drug side effects, poor treatment effect, low cure rate, high mortality rate, but also high treatment costs, which brings a heavy economic burden to patients

Method used

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  • Kit for detecting or assisting in detecting ethambutol resistance of multiple drugs resistant bacillus tuberculosis
  • Kit for detecting or assisting in detecting ethambutol resistance of multiple drugs resistant bacillus tuberculosis
  • Kit for detecting or assisting in detecting ethambutol resistance of multiple drugs resistant bacillus tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Obtaining of the ethambutol resistance kit for detection or auxiliary detection of multidrug-resistant Mycobacterium tuberculosis

[0032] 1. Acquisition of multidrug-resistant Mycobacterium tuberculosis ethambutol resistance-related gene mutation sites

[0033] (1) Genomic DNA was extracted, as follows:

[0034] The sputum samples of 61 tuberculosis patients from the Beijing Institute of Tuberculosis Control and 110 tuberculosis patients from the Shanghai Center for Disease Control and Prevention who were identified as resistant to rifampicin (RIF) and isoniazid (INH) Smear, culture, isolation, purification, etc. These tuberculosis patients were all infected with multidrug-resistant Mycobacterium tuberculosis (MDR Mycobacterium tuberculosis), and MDR-resistant Mycobacterium tuberculosis (slender and slightly curved bacilli, 1 to 4X0.4μm), save for future use;

[0035] Resuscitate the MDR Mycobacterium tuberculosis isolated and purified above in a biosafety...

Embodiment 2

[0063] Example 2, Application of the Ethambutol Resistance Kit for Detection or Auxiliary Detection of Multidrug-resistant Mycobacterium tuberculosis

[0064] The samples were: 61 tuberculosis patients identified as ethambutol-resistant and 21 tuberculosis patients identified as non-ethambutol-resistant from Beijing Institute of Tuberculosis Control, Shanghai Center for Disease Control and Prevention Sputum samples from 110 TB patients identified as ethambutol-resistant and 45 TB patients identified as non-ethambutol-resistant; these TB patients were all infected with multidrug-resistant Mycobacterium tuberculosis, That is, all patients were resistant to rifampicin (RIF) and isoniazid (INH) after testing, and all patients who underwent the following experiments were informed.

[0065] Method 1: Detection of ethambutol resistance at 4 mutation sites

[0066] 1. Separate and purify the sputum samples of each patient in the above-mentioned regions to obtain multidrug-resistant M...

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Abstract

The invention discloses a kit for detecting or assisting in detecting ethambutol resistance of multiple drugs resistant bacillus tuberculosis. The kit comprises a product for detecting an embB354 mutation site in an embB gene, wherein the nucleotide sequence of the embB gene is the sequence 1 shown in a sequence table; and the embB354 mutation site is that nucleotide GAC from the 1060th site to the 1062nd site at the 1-5' ends of the sequences in the sequence table is mutated into GCC. Tests show that the new mutation site embB354 of the embB gene can be used for detecting the ethambutol resistance of the multiple drugs resistant bacillus tuberculosis, and the mutation site embB354 is combined with another three mutation sites, namely, embB306, embB406 and embB497, to detect the ethambutol resistance of the multiple drugs resistant bacillus tuberculosis together so as to ensure higher diagnosis sensitivity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an ethambutol drug resistance kit for detecting or assisting in detecting multidrug-resistant Mycobacterium tuberculosis. Background technique [0002] Multidrug resistance (MDR) refers to the resistance of Mycobacterium tuberculosis to at least two first-line anti-tuberculosis drugs, rifampicin (RIF) and isoniazid (INH). Multidrug-resistant tuberculosis not only has long treatment cycle, serious drug side effects, poor treatment effect, low cure rate, high mortality rate, but also high treatment cost, which brings a heavy economic burden to patients. Early and rapid diagnosis of drug resistance in Mycobacterium tuberculosis is beneficial to the treatment and control of MDR-TB. Compared with the traditional drug sensitivity detection method based on solid and liquid culture, the drug sensitivity detection method based on molecular biology (molecular diagnosis of drug resistance) has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 石大伟甄伟朱国峰
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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