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Mycobacterium tuberculosis drug-resistant gene multiple detection method based on RCA (rolling circle amplification)

A Mycobacterium tuberculosis, multiple detection technology, applied in the direction of microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of time-consuming detection methods, expensive molecular biology techniques, and high operational requirements , to achieve the effect of low cost, low price, and improved detection efficiency

Pending Publication Date: 2019-12-10
张阳
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Problems solved by technology

[0003] To sum up, the problems existing in the existing technology are: the current molecular biology technology based on drug resistance gene detection is expensive, requires high operation, and the traditional detection method takes a long time.

Method used

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  • Mycobacterium tuberculosis drug-resistant gene multiple detection method based on RCA (rolling circle amplification)
  • Mycobacterium tuberculosis drug-resistant gene multiple detection method based on RCA (rolling circle amplification)
  • Mycobacterium tuberculosis drug-resistant gene multiple detection method based on RCA (rolling circle amplification)

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Embodiment Construction

[0027] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0028] The present invention constructs a simple, sensitive and fast novel POCT paper-based microfluidic analysis device for identifying drug resistance of Mycobacterium tuberculosis, and integrates isothermal nucleic acid amplification and visual color development into the paper-based microfluidic analysis device. The detection process of amplifying the sample to be tested and developing the color of the paper base under constant temperature conditions is completed within 2-4 hours, and does not require complicated instruments and equipment.

[0029] The application principle of the present invention will be described in deta...

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Abstract

The invention belongs to the technical field of gene detection and discloses a mycobacterium tuberculosis drug-resistant gene multiple detection method based on RCA (rolling circle amplification). Base sequences for detection are shown in SEQ ID NO:1 to SEQ ID NO:28. A paper based microfluidic analysis device comprises a sample pad, a colloidal gold conjugation pad, a detection line, a control line, an NC (nitrocellulose filter membrane), a water absorption pad and a bottom plate, wherein the sample pad, the colloidal gold conjugation pad, the NC and the water absorption pad are sequentially arranged on the bottom plate from left to right; and the detection line and the control line are arranged on the NC. Through massive serial-connection repeated sequence hybridization of nano gold granule modified common sequence probes and single base mutation probes with a solid-phase surface RCA reaction product, a color reaction indication detection result can be achieved. A detection platform of an isothermal amplification conjunction paper based developing sensation technique is established, and an experiment method for rapidly identifying and detecting mycobacterium tuberculosis drug resistance is established.

Description

technical field [0001] The invention belongs to the technical field of gene detection, in particular to a method for multiple detection of drug-resistant genes of Mycobacterium tuberculosis based on RCA amplification. Background technique [0002] At present, the commonly used existing technology in the industry is as follows: The "Global Tuberculosis Report" issued by the World Health Organization (WHO) shows that tuberculosis is one of the top ten causes of death in the world with the nature of air droplet transmission, and it is a serious health hazard. Chronic infectious disease is one of the infectious diseases that my country and the world focus on prevention and control. In tuberculosis, due to the increase in population and mobility, delay in treatment and irregular use of anti-tuberculosis drugs, as well as the prevalence of immunodeficiency diseases such as AIDS, the emergence of drug-resistant strains of Mycobacterium tuberculosis, the formation of multidrug-resist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/34C12M1/38C12Q1/10C12Q1/689C12Q1/6844C12R1/32
CPCB01L7/52B01L3/5027C12Q1/10C12Q1/689C12Q1/6844
Inventor 张阳夏柯府伟灵
Owner 张阳
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