Microfluidic chip and detection method for nucleic acid detection
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A technology of microfluidic chip and detection method, which can be used in laboratory containers, chemical instruments and methods, biochemical equipment and methods, etc., and can solve problems such as slow speed and unsuitability for bedside diagnosis.
Active Publication Date: 2022-03-25
INST OF URBAN SAFETY & ENVIRONMENTAL SCI BEIJING ACAD OF SCI & TECH
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[0003] Quantitative reverse transcription polymerase chain reaction is considered the "gold standard" and is often used for nucleic acid detection of new coronaviruses, but this method requires professional personnel and specialized equipment, is not only slow and not suitable for bedside diagnosis
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Embodiment 1
[0039] This embodiment provides a nucleic acid detection chip with high sensitivity and high specificity, which integrates LAMP amplification and CRISPR / Cas12a cutting system into the same microfluidic chip. Its structure is as follows: Figure 1~Figure 3 shown.
[0040] The microfluidic chip includes a three-layer structure stacked together and sealed to each other in sequence, from top to bottom are the air channel layer 1, the middle layer 2 and the flow channel layer 3;
[0041] The airway layer contains two independent airways, respectively airway I ( figure 2 middle 4), airway II ( figure 2 middle 5); wherein, the airway I is in the shape of a 1, corresponding to the position of the microvalve ④ on the flow channel layer; the airway II is in the shape of a T, and one side of the T shape is in line with the position of the microvalve ① and microvalve ② on the flow channel layer. corresponding to the positions; the other side of the T shape corresponds to the position ...
Embodiment 2
[0062] Example 2 Method for detecting nucleic acid using a microfluidic chip
[0063] Taking the detection of the new coronavirus as an example, the specific detection methods are as follows:
[0064] 1. Take the microfluidic chip made in Example 1, the airway I is blocked from gas, and the airway II is led into gas, so that the microvalve ④ is opened, and the microvalve microvalves ①, ②, and ③ are closed;
[0065] 2. Injection port ① into the SARS-COV-2 virus target RNA (0.2-0.8μl / min), injection port ② into the LAMP amplification reaction solution (0.2-0.8μl / min), the two liquids flow to LAMP Amplification chamber 12;
[0066] 3. Heat the chip at 65°C for 40 minutes;
[0067] 4. The air channel I is filled with gas, and the air channel II is blocked, so that the micro valve ④ is closed, and the micro valves ①, ②, ③ are opened;
[0068] 5. Inject double-distilled water into the injection port ③, squeeze the liquid (0.2-0.8 μl / min) of the LAMP amplification chamber 12 into ...
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Abstract
The invention provides a microfluidic chip and a detection method for nucleic acid detection. The microfluidic chip includes a three-layer structure stacked together and sealed to each other, from top to bottom are respectively an air channel layer, a middle layer and a channel layer; the air channel layer contains two independent air channels, The middle layer is an elastic film, which is used to control the opening and closing of the microvalve on the flow channel layer. The flow channel layer includes four sample inlets, two sample outlets, four microvalves, a LAMP reaction chamber, A CRISPR reaction chamber and several flow channels, the microvalve is connected to the airway of the airway layer through an elastic film, and the switch of the microvalve is realized by changing the air pressure of the airway layer, so as to realize the sequential injection of different injection ports . The invention combines microfluidics with LAMP amplification technology and CRISPR detection technology to realize highly sensitive and highly specific detection of viral nucleic acids on a single chip.
Description
technical field [0001] The invention relates to the technical field of molecular detection, in particular to a microfluidic chip and a detection method for nucleic acid detection. Background technique [0002] Novel coronavirus pneumonia (Corona Virus Disease 2019, COVID-19) is an acute respiratory infectious disease. Patients mainly present with fever, dry cough, fatigue, etc., which can lead to acute respiratory distress syndrome, septic shock, difficult to correct Metabolic acidosis, coagulation dysfunction and multiple organ failure. There is an urgent need for a fast, convenient and accurate method to detect the new coronavirus. [0003] Quantitative reverse transcription polymerase chain reaction is considered the "gold standard" and is often used for nucleic acid detection of new coronaviruses. However, this method requires professional personnel and specialized equipment, which is not only slow but also not suitable for bedside diagnosis. Clustered regularly inters...
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