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Method for identifying Mycobacterium tuberculosis and non-tuberculous mycobacteria, and special reagent kit therefor

A technology of mycobacterium tuberculosis and mycobacteria, which is applied in the direction of biochemical equipment and methods, and the determination/testing of microorganisms, and can solve problems such as delaying treatment of patients

Active Publication Date: 2013-03-20
BOAO BIOLOGICAL CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The distinction between mycobacterium tuberculosis and non-tuberculosis mycobacteria still relies on the traditional method of isolation and culture, and it usually takes 1-4 weeks to get the exact results, which greatly delays the treatment of patients and causes additional costs to patients. financial burden and mental burden

Method used

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  • Method for identifying Mycobacterium tuberculosis and non-tuberculous mycobacteria, and special reagent kit therefor
  • Method for identifying Mycobacterium tuberculosis and non-tuberculous mycobacteria, and special reagent kit therefor
  • Method for identifying Mycobacterium tuberculosis and non-tuberculous mycobacteria, and special reagent kit therefor

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1, the identification of mycobacterium tuberculosis and non-tuberculous mycobacteria

[0030] The kit for distinguishing mycobacterium tuberculosis from non-tuberculosis mycobacteria consists of the following parts: reagents for extracting nucleic acid, PCR reaction reagents, primers, probes, positive sample controls and negative sample controls, and instructions. The primers are primers P1 and P2, primers P3 and P4, the 5' end of probe 1 is labeled with FAM fluorescence, and the 3' end has an Eclipse fluorescence quencher group. The 5' end of probe 2 is labeled with HEX fluorescence, and the 3' end is equipped with Eclipse fluorescence quencher group.

[0031] Primers P1 and P2 are designed for the sequence of Mycobacterium tuberculosis IS6110, and the nucleotide sequences of primers P1 and P2 are as follows:

[0032] Primer P1: 5'-AAGCCCGCAGGACCACGAT-3' (sequence 1 in the sequence listing);

[0033] Primer P2: 5'-CGTAGGCGTCGGTGACAAAGG-3' (SEQ ID NO: 2 in ...

Embodiment 2

[0045] Embodiment 2, the identification of mycobacterium tuberculosis and non-tuberculous mycobacteria

[0046] The kit for distinguishing mycobacterium tuberculosis from non-tuberculosis mycobacteria consists of the following parts: reagents for extracting nucleic acid, PCR reaction reagents, primers, probes, positive sample controls and negative sample controls, and instructions. The primers are primers P1 and P2, primers P3 and P4, the 5' end of probe 1 is labeled with FAM fluorescence, and the 3' end has an Eclipse fluorescence quencher group. The 5' end of probe 2 is labeled with HEX fluorescence, and the 3' end is equipped with Eclipse fluorescence quencher group.

[0047]Primers P1 and P2 are designed for the MTP40 sequence of Mycobacterium tuberculosis, and the nucleotide sequences of primers P1 and P2 are as follows:

[0048] Primer P1: 5'-CAAGTTGGGTCTGGTCGAATTC-3' (sequence 7 in the sequence listing);

[0049] Primer P2: 5'-CACACCTCGCAATTGAAGTTGA-3' (SEQ ID NO: 8 i...

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Abstract

The invention discloses a method for identifying Mycobacterium tuberculosis and nontuberculosis mycobacteria and a special reagent kit thereof. The reagent kit for identifying the Mycobacterium tuberculosis and the nontuberculosis mycobacteria comprises a primer and a probe, wherein the primer comprises four primers, and ribonucleotide sequences of the four primers are sequence 1, sequence 2, sequence 4, and sequence 5 in a sequence table respectively; the probe comprises two probes, and ribonucleotide sequences of the two probes are sequence 3 and sequence 6 in the sequence table respectively; 5' ends of the probes are provided with fluorescent substances, and 3' ends of the probes are provided with quenching substances; and the fluorescent substances at the 5' ends of the two probes are different. The reagent kit has the advantages that the reagent kit can quickly identify the Mycobacterium tuberculosis and the nontuberculosis mycobacteria, has simple requirement on instruments, low cost and simple operation, and has broad application prospect on early diagnosis and differential diagnosis of tuberculosis.

Description

technical field [0001] The invention relates to a method for distinguishing mycobacterium tuberculosis and non-tuberculosis mycobacterium and a special kit thereof. Background technique [0002] Tuberculosis (Tuberculosis) is one of the most threatening infectious diseases in the world today. Since the mid-1980s, there has been a global recovery trend. According to the report of the World Health Organization, my country is one of the 22 countries with a high burden of tuberculosis in the world. The number of active tuberculosis patients ranks second in the world. At present, the tuberculosis infection rate of all age groups in my country is 44.5%, and about 550 million people in the country have been infected by tuberculosis bacteria. The tuberculosis infection rate is higher than the global population infection rate of 1 / 3. At the same time, non-tuberculous mycobacteria (NTM) disease is also on the rise year by year. The flow survey data organized by the Ministry of Health ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 王思贤侯伟高华方程京
Owner BOAO BIOLOGICAL CO LTD
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