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Primers and kit for quickly detecting Mycobacterium tuberculosis isoniazide drug resistance

A technology of Mycobacterium tuberculosis and isoniazid, applied in the field of molecular biology, can solve problems such as time-consuming

Inactive Publication Date: 2016-09-28
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method takes a long time, generally takes 1-2 months, and some severe patients died before the results of drug sensitivity came out

Method used

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  • Primers and kit for quickly detecting Mycobacterium tuberculosis isoniazide drug resistance
  • Primers and kit for quickly detecting Mycobacterium tuberculosis isoniazide drug resistance
  • Primers and kit for quickly detecting Mycobacterium tuberculosis isoniazide drug resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Materials and methods

[0049] 1. Materials

[0050] 1.1 Source of strains: The 40 strains used in this experiment were isolated and preserved in our laboratory, among which 1 strain was the standard strain of Mycobacterium tuberculosis H37Rv, 21 strains were isoniazid-resistant strains, and 28 strains were isoniazid-susceptible strains.

[0051] 1.2 Main instruments and equipment: real-time fluorescent quantitative PCR instrument ( Roche, Switzerland), vortex mixer (VORTEX-3, Shanghai Yesterday Fei Laboratory Equipment Co., Ltd.)

[0052] 1.3 Reagents and media:

[0053] (1) DNA extraction kit: Tianze Gene Column Mycobacterium DNAout (produced by Beijing Tianenze), high-resolution melting curve reaction reagent (produced by Roche, Switzerland)

[0054] (2) Primers:

[0055] katG Forward: 5'—TCGTATGGCACCGGAACC—3'

[0056] katG Reverse: 5'—CAGCTCCCACTCGTAGCC—3'

[0057] katG-1Forward: 5’—GGGCTGGAAGAGCTCGTAT—3’

[0058] katG-1Reverse: 5’—CCGTACAGGATCTCG...

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Abstract

The invention discloses primers and a kit for quickly detecting Mycobacterium tuberculosis isoniazide drug resistance. The primers comprises at least one set of the following primers: first set: katG Forward: 5'-TCGTATGGCACCGGAACC-3' and katG Reverse: 5'-CAGCTCCCACTCGTAGCC-3'; second set: katG-1 Forward: 5'-GGGCTGGAAGAGCTCGTAT-3' and katG-1 Reverse: 5'-CCGTACAGGATCTCGAGGAA-3'; and third set: inhA Forward: 5'-CGTTACGCTCGTGGACATAC-3' and inhA Reverse: 5'-TCCGGTAACCAGGACTGAAC-3'.

Description

technical field [0001] The invention relates to the research and development of a biological detection reagent, in particular to a primer and a kit for rapid detection of isoniazid resistance of Mycobacterium tuberculosis, belonging to the technical field of molecular biology. Background technique [0002] Isoniazid was synthesized as early as 1912. It was discovered in 1952 that it has a strong antibacterial and bactericidal effect on Mycobacterium tuberculosis. The drug is completely absorbed in the digestive tract and easily penetrates into lung tissue and cerebrospinal fluid. . After being acetylated and hydroxylated in the liver, isoniazid can be excreted with urine and has low toxicity. Due to its strong effect, good curative effect, small dosage and low price, isoniazid has become a clinical anti-tuberculosis drug for many years. drug of choice. Decades have passed, and Mycobacterium tuberculosis has developed resistance to isoniazid. According to the data of the t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/689C12Q1/6827C12Q2600/156C12Q2527/107
Inventor 曹旭东杨敏杨彩虹于潞陈创夫袁俐吴长新
Owner SHIHEZI UNIVERSITY
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