Power source-free and valve-free type single molecule detection chip and applications thereof

A single-molecule detection, valveless technology, used in laboratory containers, microbial determination/inspection, bioreactor/fermenter combination, etc. Many other problems, to achieve the effect of improving the speed of the experiment, reducing the consumption and reducing the cost of the experiment

Active Publication Date: 2013-05-01
杭州三济生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Routine nucleic acid amplification experiments are carried out using small test tubes, orifice plates, etc. The workload is heavy, and there are many steps to add samples, which is prone to impurity contamination
In addition, traditional PCR needs to be opened for detection, resulting in aerosol pollution, which makes the false positive rate very high (30~40%)

Method used

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  • Power source-free and valve-free type single molecule detection chip and applications thereof
  • Power source-free and valve-free type single molecule detection chip and applications thereof
  • Power source-free and valve-free type single molecule detection chip and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] see figure 1 , a microfluidic chip device for nucleic acid amplification that does not require an external drive system, does not require valves, has a simple manufacturing process and is easy to operate. The device consists of a reaction layer 1, a modification layer 2, a sealing layer 3, a main channel 4, a reaction chamber 5, a sample inlet 6, a sample outlet 7, and a heat-resistant tape 8. The reaction layer 1 is composed of a main channel 4 in series with reaction chambers. 5, the reaction chamber 5 is located above the main channel 4, the sample inlet 6 and the sample outlet 7 are located at both ends of the main channel 4, the modification layer 2 is connected to the sealing layer 3 in advance, and the reaction layer 1 is connected to the sealing layer through the modification layer 2. 3 After sealing, the microfluidic chip assembly is formed. A heat-resistant adhesive tape 8 is pasted on the upper surface of the reaction layer 1 .

[0057] The reaction layer 1...

Embodiment 2

[0068] see figure 2 , a microfluidic chip device for nucleic acid amplification that does not require an external drive system, does not require valves, has a simple manufacturing process and is easy to operate. The device consists of a reaction layer 1, a modification layer 2, a sealing layer 3, a main channel 4, a reaction chamber 5, a sample inlet 6, a sample outlet 7, and a heat-resistant tape 8. The reaction layer 1 is composed of a main channel 4 in series with reaction chambers. 5, the reaction chamber 5 is located below the main channel 4, the sample inlet 6 and the sample outlet 7 are located at both ends of the main channel 4, the modification layer 2 is connected to the sealing layer 3 in advance, and the reaction layer 1 is connected to the sealing layer through the modification layer 2. 3 After sealing, the microfluidic chip assembly is formed. A heat-resistant adhesive tape 8 is pasted on the upper surface of the reaction layer 1 . The reaction layer 1 is made...

Embodiment 3

[0078] Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) on a valve-free microfluidic PDMS chip without power source.

[0079] In this example, the device in Example 1 is used to inject samples, and the subsequent real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) is performed. Specific steps:

[0080] 1. See image 3 Channel structures are designed and masked.

[0081] 2. The reaction layer 1 is made of polydimethylsiloxane (PDMS), and a substrate with a microchannel structure produced by multi-layer soft lithography technology is used as a mold, and PDMS with gas permeability is poured on the mold and solidified. The reaction chamber 5 is a square with a side length of 100 μm, 200 μm or 300 μm, and a depth of 50 μm, 100 μm, 200 μm, or 300 μm. According to different widths and mold heights, the volume of the reaction chamber 5 can be 500 pL, 1 nL, 2 nL, 4.5 nL, 8 nL, 9 nL, 18 nL; the reaction chamber 5 can also be penta...

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Abstract

The present invention provides a power source-free and valve-free type single molecule detection chip, which comprises a reaction layer, a modification layer, a sealing layer, a main channel, small reaction chambers, a sample inlet, a sample outlet and a heat resistant adhesive tape, wherein the reaction layer is formed by connecting the main channel and the reaction small chambers in series, the reaction small chambers are positioned above or below the main channel, the sample inlet and the sample outlet are positioned on both ends of the main channel, the modification layer is pre-connected with the sealing layer, the reaction layer is sealed with the sealing layer through the modification layer to form a microfluidic chip assembly, and the heat resistant adhesive tape is adhered on the upper surface of the reaction layer. According to the present invention, miniaturization is achieved, such that consumption of reagents and samples are reduced, and cost is low; a small amount of liquid can be assigned to hundreds of the independent small chambers without an external connection drive system and valve switching control equipment so as to reduce complexity of chip production and use; and the operation is simple, other control devices are not required, application cost is low, and the chip can be used in real-time fluorescence quantitative polymerase chain reactions and digital nucleic acid amplification.

Description

technical field [0001] The invention belongs to detection devices in multiple fields such as life sciences and medicine, and relates to a microfluidic chip device for nucleic acid amplification and its application technology. Background technique [0002] Major diseases such as cancer, cardiovascular and cerebrovascular diseases, and infectious diseases claim the lives of millions of people in our country every year. According to the announcement of the Ministry of Health in 2010, there are 6.6 million cancer patients and 1.8 million deaths in my country every year. The annual medical expenses for cancer patients are as high as hundreds of billions of RMB. As the population ages, it is estimated that by 2020, there will be 3 million new cancer cases and 3 million deaths. Cancer has become a heavy burden of disease in our country, and it is very important to strengthen the research of new methods and new technologies for cancer prevention and early diagnosis and treatment. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00B01J19/00C12M1/34C12Q1/68
Inventor 牟颖朱强远高一搏金伟
Owner 杭州三济生物科技有限公司
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