The subject matter relates to nucleic acid detection devices, and more particularly, to a nucleic acid detection kit and a nucleic acid detection device with the nucleic acid detection kit.
Molecular diagnosis, morphological detection, and immunological detection are mostly carried out in laboratories. The detection process includes performing a polymerase chain reaction (PCR) amplification reaction in a large and medium-sized detection equipment to acquire an amplified product. Then, the amplified product is manually transferred to an electrophoresis detection equipment for an electrophoretic detection. Finally, an electrophoretic detection result is manually transferred to a fluorescence analyzer to obtain a fluorescence image. However, such detection process is time-consuming, inefficient, and inflexible, and the detection device is not portable. The detection cannot be carried out anytime and anywhere.
Implementations of the present technology will now be described, by way of example only, with reference to the attached figures.
It will be appreciated that for simplicity and clarity of illustration, where appropriate, reference numerals have been repeated among the different figures to indicate corresponding or analogous components. In addition, numerous specific details are set forth in order to provide a thorough understanding of the embodiments described herein. However, it will be understood by those of ordinary skill in the art that the embodiments described herein can be practiced without these specific details. In other instances, methods, procedures, and components have not been described in detail so as not to obscure the related relevant feature being described. Also, the description is not to be considered as limiting the scope of the embodiments described herein. The drawings are not necessarily to scale, and the proportions of certain parts may be exaggerated to better illustrate details and features of the present disclosure.
The term “comprising,” when utilized, means “including, but not necessarily limited to”; it specifically indicates open-ended inclusion or membership in the so-described combination, group, series, and the like.
Referring to
In an embodiment, the first housing 11 and the second housing 12 are clamped together. The first housing 11 and the second housing 12 are further fastened together by screws to increase a connection strength therebetween.
Referring to
In an embodiment, the kit body 1 may be made of, but not limit to plastic.
Referring to
Referring to
In an embodiment, the first dielectric layer 26 and the second dielectric layer 27 are insulating and hydrophobic layers. On the one hand, the first dielectric layer 26 and the second dielectric layer 27 has the characteristics of insulation and hydrophilicity, and on the other hand, the first dielectric layer 26 and the second dielectric layer 27 can make the microbead 6 to move more smoothly in the moving path and avoid breakage of the microbead 6 during movement.
In an embodiment, each of the first dielectric layer 26 and the second dielectric layer 27 may be but not limit to a polytetrafluoroethylene coating.
Referring to
In an embodiment, the control electrodes 242 are integrated at an edge of the second cover plate 23. An electrical connection between the detection chip 2 and the connector 4 is realized by inserting the side of the second cover plate 23 with the control electrodes 242 into the connector 4.
Referring to
A principle of real-time fluorescence quantitative PCR technology is that the fluorescent reagent (a fluorescent dye or a DNA probe) is designed to have fluorescence characteristics only after the fluorescent reagent is combined with a DNA. Therefore, when the number of the DNA increase after the PCR amplification reaction, and more fluorescent reagents are activated after combing with the DNAs, and a stronger fluorescence intensity may be obtained. The amplified DNAs can be quantified by detecting the fluorescence intensity.
Referring to
Referring to
Referring to
In an embodiment, the heating unit 28 includes two heating areas. Each of the two heating areas corresponds to a PCR amplification area “C”. One of the two heating areas has a heating temperature ranges from 90° C. to 105° C. The other one of the two heating areas has a heating temperature ranges from 40° C. to 75° C.
In an embodiment, silicone oil may be injected into the channel 5 after the detection chip 2 is assembled, and the microbead 6 is driven to move in the silicone oil.
Referring to
In an embodiment, the nucleic acid detection kit 100 is substantially cubic.
In an embodiment, the nucleic acid detection kit 100 is disposable. The nucleic acid detection kit 100 has no need to be cleaned after used.
The numbers and the positions of the PCR amplification area “C”, the reagent storage area “B”, and the observation area “D” can be designed according to different needs. Three different embodiments in an actual detection process are illustrated as follows.
Referring to
A process of performing the real-time fluorescence quantitative PCR includes flowing steps.
At step one, the microbead 6 is added into the sampling area “A” through the sampling port 13. The microbead 6 includes the nucleic acid sample, the primer, and the fluorescent reagent (such as a fluorescent dye or a DNA probe).
At step two, the detection solution 6 is driven to move back and forth between the two PCR amplification areas “C” along the moving path. During this process, the nucleic acid sample and the primer are heated to undergo the PCR amplification reaction to form the amplified product, then the amplified product combined with the fluorescent reagent to form the mixer microbead 8 at the same time. During the PCR amplification reaction process, the mixer microbead 8 may pass through the observation area “D”, so that the mixer microbead 8 may send out fluorescent signal 9 under the irradiation of the laser beam 7. Then, the image collection unit can collect the fluorescence signal 9 at the observation area “D” through the observation window 29. The image collection unit may output a fluorescent image to a computer to calculate the fluorescence intensity. The fluorescence intensity increases continuously during the progress of the PCR amplification reaction. When the fluorescence intensity reaches a maximum value which tends to be stable, and then the PCR amplification reaction is ended. Therefore, an end time of the PCR amplification reaction can be accurately determined according to the change of the fluorescence intensity.
In an embodiment, the laser emitter 3 emits the laser beam 7 from a side of the channel 5, and the laser beam 7 is transmitted in the channel 5. In an embodiment, a spectrum of the laser beam 7 is ranges from 200 nm to 480 nm. A spectrum of a fluorescence emitted by the fluorescent reagent is ranges from 500 nm to 700 nm. When the laser beam 7 intersects with the mixer microbead 8, the fluorescent reagent activated in the mixer microbead 8 sends out a fluorescent signal 9. The image collection unit detects and collects the fluorescent signal 9 through the opening 14 and the observation window 29 to form a fluorescent image, which is then sent to the computer.
Referring to
A process of realizing the real-time fluorescence quantitative PCR includes flowing steps.
At step one, add the microbead 6 into the sampling area “A” through the sampling port 13. The microbead 6 contains the nucleic acid sample and the primer. The fluorescent reagent (such as a fluorescent dye or a DNA probe) is coated in the reagent storage area “B”.
At step two, the detection solution 6 is driven to move from the sampling area “A” to the reagent storage area “B” to mixed with the fluorescent reagent to form a mixer.
At step three, the mixer is driven to move back and forth between the two PCR amplification areas “C” along the moving path. During this process, the nucleic acid sample and the primer are heated to undergo the PCR amplification reaction to form the amplified product, then the amplified product combined with the fluorescent reagent to form the mixer microbead 8 at the same time. During the PCR amplification reaction process, the mixer microbead 8 may pass through the observation area “D”, so that the mixer microbead 8 may send out fluorescent signal 9 under the irradiation of the laser beam 7. Then, the image collection unit can collect the fluorescence signal 9 at the observation area “D” through the observation window 29. The image collection unit may output a fluorescent image to a computer to calculate the fluorescence intensity. The fluorescence intensity increases continuously during the progress of the PCR amplification reaction. When the fluorescence intensity reaches a maximum value which tends to be stable, and then the PCR amplification reaction is ended. Therefore, an end time of the PCR amplification reaction can be accurately determined according to the change of the fluorescence intensity.
Referring to
In an embodiment, three different DNA probes are set on the three observation sites.
In an embodiment, a RdRp gene probe is set on the observation site “D1”. N gene probe is set on the observation site “D2”, and Beta actin gene probe is set on the observation site “D3”.
A process of realizing the real-time fluorescence quantitative PCR includes flowing steps.
At step one, add the microbead 6 into the sampling area “A” through the sampling port 13. The microbead 6 contains the nucleic acid sample and the primer.
At step two, the detection solution 6 is driven to move back and forth between the two PCR amplification areas “C” along the moving path. During this process, the nucleic acid sample and the primer are heated to undergo the PCR amplification reaction to form the amplified product.
At step three, the amplified product is driven to the observation area “D” to combine with the three fluorescent reagents to form three mixture microbeads 8. The three mixture microbeads 8 may send out three fluorescent signals 9 under the irradiation of the lasers 7. Then, the image collection unit can collect the fluorescence signals 9 at the observation area “D” through the observation window 29. The image collection unit may output a photo which contain three fluorescent images to a computer to calculate the fluorescence intensities of the three fluorescent images.
In order to determine the effect of the fluorescent reagent added before or after the PCR amplification reaction, three fluorescence detections are carried out. Referring to
With the above configuration, the nucleic acid detection kit 100 can realize the real-time fluorescence quantitative PCR in a single equipment through the cooperation of the detection chip 1 and the laser emitter 3. The mixer microbead 8 is undergo the fluorescence detection after the PCR amplification reaction in the detection chip 1 to form the fluorescence image. The mixer microbead 8 does not need to be undergo an electrophoretic detection. Thus, the nucleic acid detection kit 100 has a simple structure, which is low cost and highly efficient.
With the above configuration, the nucleic acid detection device 200 can realize the real-time fluorescence quantitative PCR in a single equipment through the cooperation of the nucleic acid detection host 201, the nucleic acid detection kit 100, and the image collection unit 202. According to the fluorescence intensity, the amount of the nucleic acid in the mixer microbead 8 can be detected quantitatively in real time. Thus, the nucleic acid detection device 200 has a simple structure, which is low cost, highly efficient, portable, flexible, and convenient, and can be used at home to realize a real time fluorescence detection. At the same time, the detecting process is flexible, which does not need to be carried out in a professional laboratory.
The embodiments shown and described above are only examples. Even though numerous characteristics and advantages of the present technology have been set forth in the foregoing description, together with details of the structure and function of the present disclosure, the disclosure is illustrative only, and changes may be made in the detail, including in matters of shape, size and arrangement of the parts within the principles of the present disclosure, up to and including, the full extent established by the broad general meaning of the terms used in the claims.
Number | Date | Country | Kind |
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202110693498.4 | Jun 2021 | CN | national |
Number | Date | Country | |
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63085368 | Sep 2020 | US | |
63140466 | Jan 2021 | US |