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Nucleic acid isothermal amplification method based on hairpin structure and application of kit

A constant temperature amplification, hairpin technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of instrument and equipment dependence, time-consuming and laborious, complicated steps, etc., and achieve rapid amplification of target sequences. Effect

Pending Publication Date: 2020-12-04
南京达伯可特生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0008] The present invention aims at the deficiencies of time-consuming and laborious nucleic acid molecular biology detection in the field of biotechnology, cumbersome steps and dependence on instruments and equipment, and aims to provide people with a simple and cheap nucleic acid constant temperature detection method, so that Advanced molecular detection methods can be widely promoted and applied effectively in on-site detection events and grassroots community hospitals

Method used

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  • Nucleic acid isothermal amplification method based on hairpin structure and application of kit
  • Nucleic acid isothermal amplification method based on hairpin structure and application of kit
  • Nucleic acid isothermal amplification method based on hairpin structure and application of kit

Examples

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Embodiment 1

[0077] Embodiment 1. Hepatitis B virus detection kit

[0078] The pathogenic factor of hepatitis B virus is hepatitis B virus, abbreviated as HBV, and hepatitis B virus is a DNA virus. The genome is double-stranded, circular, partially closed DNA. The outermost layer of the virus is the outer membrane or envelope of the virus, and the inner layer is the core. The nucleoprotein is the core antigen (HBcAg), which cannot be detected in serum. Hepatitis B virus is an infectious disease mainly caused by HBV, mainly liver lesions. Clinically, the main manifestations are loss of appetite, nausea, upper abdominal discomfort, pain in the liver area, and fatigue. Some patients may have jaundice, fever and hepatomegaly with liver function damage. Some patients can become chronic and even develop into liver cirrhosis, and a few can develop into liver cancer. HBV is an infectious disease that seriously endangers human health. It is a major problem that hinders the country's economic an...

Embodiment 2

[0101] Example 2. HBV amplification product detection

[0102] In the reaction system of Example 1, Eva Green was replaced with calcein and other reaction conditions were kept unchanged, and then the constant temperature amplification products were detected with a hand-held ultraviolet light source and a gel electrophoresis system respectively. The test results show that the Figure 4 in B.

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Abstract

The invention relates to a method for designing a related specific primer, so that a hairpin structure with special purposes is formed when a target nucleic acid sequence is amplified under the isothermal condition, and the hairpin structure mediates subsequent exponential amplification, so that the purpose of realizing efficient and high-sensitivity detection on target molecules is achieved. Meanwhile, the invention also relates to application of a related kit in the aspect of rapid nucleic acid diagnosis, including application in the aspect of detection of pathogenic microorganisms such as bacteria and viruses.

Description

technical field [0001] The invention relates to a nucleic acid molecular biology detection method in the field of biotechnology, in particular to a nucleic acid constant temperature amplification method based on a hairpin structure and the application of a kit thereof. Background technique [0002] The investigation of the global epidemic trend shows that conventional pathogen detection methods, such as bacterial culture, morphological detection and biochemical identification, etc., have been unable to be effective due to their low sensitivity, poor specificity, cumbersome steps and dependence on instruments and equipment. It meets the requirements of rapid and specific on-site detection of pathogenic microorganisms. The rapid development of molecular biology technology, especially nucleic acid amplification technology, has greatly improved the detection level of pathogenic bacteria, so it has a good control and prevention effect on the spread and treatment of infectious dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6844
CPCC12Q1/6844C12Q2525/301C12Q2531/119
Inventor 徐高连祁小飞侯建华单洪波
Owner 南京达伯可特生物科技有限公司