Sample pretreatment and amplification integrated nucleic acid analysis system based on pipette tip and application of system

A sample pretreatment and nucleic acid analysis technology, which is applied in the field of biotechnology and nucleic acid analysis and detection, can solve the problems of rapid detection and analysis not applicable to grassroots units, complicated sample pretreatment, expensive instruments and equipment, etc., and achieves low price, easy operation, and operation convenient effect

Active Publication Date: 2015-05-06
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing nucleic acid detection and analysis system that integrates sample pretreatment mainly has disadvantages such as complicated sample pretreatment, expensive instruments and equipment required for amplification, and professional operation, so it is not suitable for rapid detection and analysis at grassroots units

Method used

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  • Sample pretreatment and amplification integrated nucleic acid analysis system based on pipette tip and application of system
  • Sample pretreatment and amplification integrated nucleic acid analysis system based on pipette tip and application of system
  • Sample pretreatment and amplification integrated nucleic acid analysis system based on pipette tip and application of system

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Experimental program
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Effect test

Embodiment 1

[0050] figure 1 A schematic structural diagram of a nucleic acid analysis system integrating sample pretreatment and amplification based on a pipette tip is shown, figure 2 A workflow diagram of the nucleic acid analysis system is shown.

[0051] The whole analysis of nucleic acid in this implementation mainly includes the following steps:

[0052] (1) Pretreatment of samples

[0053] Samples mainly include solid tissue samples and liquid samples. For liquid samples, it can be sucked directly with a pipette; for solid tissue samples, it can be ground with a hand-held tissue grinder, and ground into interstitial fluid that can be sucked with a pipette for later use.

[0054] A pipette combined with a pipette tip integrated with a nucleic acid extraction card is used to aspirate the sample, and the liquid sample contacts the nucleic acid extraction card. After waiting for a certain period of time to complete the automatic extraction of nucleic acid on the card, blow out the...

Embodiment 2

[0060] In this implementation, the nucleic acid analysis system is used for the detection of potato pathogenic bacteria in inspection and quarantine, and the specific steps are as follows:

[0061] Firstly, the tissue at the suspicious spot on the plant tissue should be processed, including cutting off the tissue, collecting it into a sample bag, and registering the number; then using a tissue grinder to grind the registered tissue, and the processed tissue The tissue grinding solution is stored for later use.

[0062] Use the pipette tip in Example 1 to draw 2 μL of tissue grinding liquid to contact the nucleic acid extraction card, let it stand for 30 minutes, and complete the extraction of nucleic acid; blow out the waste liquid, and suck the cleaning solution to clean the sample, and keep it on the nucleic acid extraction card. A relatively pure nucleic acid is obtained.

[0063] The pipette tip is used to draw the nucleic acid amplification reaction solution containing t...

Embodiment 3

[0066] In this embodiment, the nucleic acid analysis system is used for the detection and analysis of multi-throughput human drug metabolism genes, and the main steps are as follows:

[0067] This system can be combined with a multi-channel pipette to detect and analyze two single-base mutation sites of CYP2C19*2 and CYP2C19*3 in the human drug metabolism gene CYP2C19 gene. The specific steps are as follows: (1) Number the two collected samples as 1# and 2# respectively, divide them into 4 parts, and use an 8-channel pipette for simultaneous detection and analysis of double parts, wherein each of the four channels Test a sample. Aspirate 2μL sample for nucleic acid extraction process; (2) For the four channels of a sample, respectively aspirate the nucleic acid amplification solution containing the wild-type and mutant-specific primers for the drug metabolism CYP2C19 gene *2 and *3 mutation sites, and then The tip is packaged, and the tip is placed on a heating platform for h...

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Abstract

The invention relates to a sample pretreatment and amplification integrated nucleic acid analysis system based on a pipette tip and the application of the system. According to the system, a nucleic acid extraction card integrated on the pipette tip is mainly used for pretreating the sample, and the nucleic acid is extracted by use of a solid-phase extraction method; next, a hand-held heating table is utilized to perform nucleic acid amplification on a nucleic acid reaction liquid; in addition, an ultraviolet lamp is used for visually judging and reading the result of detection by use of naked eyes; as a result, the real-time detection and the field detection of the nucleic acid sample are realized. The nucleic acid analysis system can be widely applied to nucleic acid detection and analysis in the fields of environmental detection, disease diagnosis, inspection andquarantine and medicolegal expertise.

Description

technical field [0001] The invention relates to the fields of biotechnology and nucleic acid analysis and detection, in particular to a pipette tip-based nucleic acid detection and analysis system integrating sample pretreatment, amplification and detection and its application in nucleic acid analysis. Background technique [0002] Molecular diagnostic technology based on nucleic acid detection and analysis has been widely used in clinical analysis and detection, providing strong technical support for disease diagnosis, prevention and prognosis evaluation. In recent years, a variety of nucleic acid detection and analysis methods have been developed, but most of the existing nucleic acid detection and analysis methods are for nucleic acid amplification detection of processed samples, which cannot achieve rapid on-site analysis and detection. [0003] In recent years, with the rapid development of the economy and the rapid increase of population flow, the frequency and probabi...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12Q1/68
CPCB01L7/525C12Q1/6806
Inventor 蒋兴宇逯文晶孙佳姝王纪东张璐查瑞涛
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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