Primer group, kit and method for simultaneously detecting mycobacterium tuberculosis composite flora and rpoB gene mutation by LAMP

A technology of complex flora and Mycobacterium tuberculosis, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unfavorable results, cumbersome separate detection operations, etc., to achieve simple operation, avoid false negatives, adapt to good sex effect

Pending Publication Date: 2020-12-01
河南中检食安生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the prior art, the Mycobacterium tuberculosis flora and the resistance of Mycobacterium tuberculosis to rifampicin are detected separately, and the separate detection operation is cumbersome, which is not conducive to rapid detection results

Method used

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  • Primer group, kit and method for simultaneously detecting mycobacterium tuberculosis composite flora and rpoB gene mutation by LAMP
  • Primer group, kit and method for simultaneously detecting mycobacterium tuberculosis composite flora and rpoB gene mutation by LAMP
  • Primer group, kit and method for simultaneously detecting mycobacterium tuberculosis composite flora and rpoB gene mutation by LAMP

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Experimental program
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Embodiment 1

[0035] The primer set and the reaction system contained in the kit for simultaneous detection of Mycobacterium tuberculosis complex flora and rpoB gene mutation by LAMP in this embodiment are as follows:

[0036] Detection of Mycobacterium tuberculosis complex

[0037] The nucleic acid detection of Mycobacterium tuberculosis complex uses the conserved IS6110 gene fragment, and the sequence of the IS6110 gene fragment is shown in SEQ ID NO:1.

[0038] The primers used for isothermal amplification of the nucleic acid of the Mycobacterium tuberculosis complex IS6110 gene are shown in Table 1.

[0039] Table 1 Primers for specific isothermal amplification of Mycobacterium tuberculosis complex

[0040] Primer name Primer sequence F3 cgccgccaactacggt B3 cggcgctggacgagat LF tcacggttcagggttagcc LB caaagcccgcaggacca FIP gcatctggccacctcgatgctacggtgcccgcaaagt BIP acggctgatgaccaaactcggcggctgtggccggatca

[0041] Table 2 shows the re...

Embodiment 2

[0109] The method for simultaneously detecting the Mycobacterium tuberculosis complex flora and the rpoB gene mutation by the LAMP of the present embodiment comprises the following steps:

[0110] 1) Extract the DNA of the purified sample by conventional methods.

[0111] 2) Preparation of the reaction system for the specific constant temperature amplification of the Mycobacterium tuberculosis compound flora, the constant temperature amplification reaction system of the 516 codon wild type of the Mycobacterium tuberculosis rpoB gene, and the constant temperature amplification reaction system of the 526 codon wild type of the Mycobacterium tuberculosis rpoB gene , Mycobacterium tuberculosis rpoB gene codon 531 wild-type constant temperature amplification reaction system, internal reference gene nucleic acid constant temperature amplification reaction system, loop-mediated isothermal amplification reaction, the reaction is carried out in an integrated device, and five sets of amp...

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Abstract

The invention relates to a primer group, a kit and a method for simultaneously detecting mycobacterium tuberculosis composite flora and rpoB gene mutation by LAMP. The primer group comprises a mycobacterium tuberculosis composite flora IS6110 gene amplification primer and an rpoB gene amplification primer group; the sequences of F3, B3, LF, LB, FIP and BIP of the mycobacterium tuberculosis composite flora IS6110 gene amplification primers are sequentially shown as SEQ ID NO: 4 to SEQ ID NO: 9; the rpoB gene amplification primer group comprises one or more of gene rpoB codon 516 wild type amplification primers, gene rpoB codon 526 wild type amplification primers and gene rpoB codon 531 wild type amplification primers; the F3, B3, FIP and BIP sequences of the gene rpoB codon 516 wild type amplification primers are sequentially shown as SEQ ID NO: 10 to SEQ ID NO: 13, the F3, B3, FIP and BIP sequences of the gene rpoB codon 526 wild type amplification primers are sequentially shown as SEQID NO: 14 to SEQ ID NO: 17, and the F3, B3, FIP and BIP sequences of gene rpoB codon 531 wild type amplification primers are sequentially shown as SEQ ID NO: 18 to SEQ ID NO: 21.

Description

technical field [0001] The invention belongs to the technical field of tuberculosis and drug-resistant gene mutation detection, and in particular relates to a primer set, a kit and a method for simultaneously detecting Mycobacterium tuberculosis complex flora and rpoB gene mutation by LAMP. Background technique [0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis, Mycobacterium bovis and Mycobacterium africanum, etc. It can affect various organs of the body, and tuberculosis is the most common. Mycobacterium tuberculosis is mainly transmitted through the air, and most people are asymptomatic after being infected with Mycobacterium tuberculosis, which is called latent infection. Latent infection can last for decades, and only 5%-10% of latent infection develop into active tuberculosis. Since the advent of therapeutic drugs one after another, tuberculosis has been basically cured. In recent years, the incidence of tuberculosis has been on...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12N15/11C12R1/32
CPCC12Q1/6844C12Q1/689C12Q2600/156C12Q2531/119
Inventor 邵俊影王云龙王群智王欣林圣博李强王丽王雯丽王昱堃
Owner 河南中检食安生物科技有限公司
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