CN113214974B - High-flux isothermal amplification one-stop detection device and use method - Google Patents
High-flux isothermal amplification one-stop detection device and use method Download PDFInfo
- Publication number
- CN113214974B CN113214974B CN202110490627.XA CN202110490627A CN113214974B CN 113214974 B CN113214974 B CN 113214974B CN 202110490627 A CN202110490627 A CN 202110490627A CN 113214974 B CN113214974 B CN 113214974B
- Authority
- CN
- China
- Prior art keywords
- sample
- detection
- hole
- value
- placing surface
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 290
- 238000000034 method Methods 0.000 title claims abstract description 55
- 238000011901 isothermal amplification Methods 0.000 title claims abstract description 50
- 238000006243 chemical reaction Methods 0.000 claims abstract description 29
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 20
- 230000008569 process Effects 0.000 claims abstract description 20
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 19
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 19
- 238000010438 heat treatment Methods 0.000 claims description 100
- OBRMNDMBJQTZHV-UHFFFAOYSA-N cresol red Chemical compound C1=C(O)C(C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C=C(C)C(O)=CC=2)=C1 OBRMNDMBJQTZHV-UHFFFAOYSA-N 0.000 claims description 17
- 238000010223 real-time analysis Methods 0.000 claims description 13
- 238000005192 partition Methods 0.000 claims description 11
- 238000001816 cooling Methods 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000013612 plasmid Substances 0.000 claims description 4
- 238000011426 transformation method Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 2
- 238000004458 analytical method Methods 0.000 abstract description 8
- 239000000523 sample Substances 0.000 description 188
- 238000007397 LAMP assay Methods 0.000 description 15
- 241000589877 Campylobacter coli Species 0.000 description 6
- 238000010586 diagram Methods 0.000 description 5
- 238000004364 calculation method Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 3
- 229910052782 aluminium Inorganic materials 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000003321 amplification Effects 0.000 description 2
- 230000007797 corrosion Effects 0.000 description 2
- 238000005260 corrosion Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 238000012123 point-of-care testing Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 241000589876 Campylobacter Species 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 238000010831 paired-sample T-test Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000007793 ph indicator Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 230000011218 segmentation Effects 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T7/00—Image analysis
- G06T7/0002—Inspection of images, e.g. flaw detection
- G06T7/0012—Biomedical image inspection
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T7/00—Image analysis
- G06T7/10—Segmentation; Edge detection
- G06T7/11—Region-based segmentation
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T7/00—Image analysis
- G06T7/90—Determination of colour characteristics
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T2207/00—Indexing scheme for image analysis or image enhancement
- G06T2207/10—Image acquisition modality
- G06T2207/10056—Microscopic image
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Organic Chemistry (AREA)
- Theoretical Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Physics & Mathematics (AREA)
- Computer Vision & Pattern Recognition (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Medical Informatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Radiology & Medical Imaging (AREA)
- Quality & Reliability (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a high-flux isothermal amplification one-stop detection device and a use method thereof, wherein the high-flux isothermal amplification one-stop detection device comprises a detection cassette, camera equipment, a cloud storage system and a computer system; the detection cassette comprises a cassette body, a first placing surface is arranged on one side of the cassette body, a second placing surface is arranged on one side of the cassette body opposite to the first placing surface, the first placing surface and the second placing surface are obliquely arranged, and a first support is arranged on the first placing surface. The high-flux isothermal amplification one-stop detection device can realize high-flux one-stop detection of a nucleic acid isothermal amplification reaction process, a sample shooting process and a detection result analysis process of LAMP detection, has a simple structure, small volume and high detection efficiency, and the use method of the high-flux isothermal amplification one-stop detection device can realize high-flux instant detection in a short time, and is convenient to operate and rapid in detection.
Description
Technical Field
The invention relates to the technical field of gene detection, in particular to a high-throughput isothermal amplification one-stop detection device and a use method thereof.
Background
Compared with the traditional PCR technology, the loop-mediated isothermal amplification (LAMP) technology has lowered the threshold of detection work, the research on LAMP detection method and detection equipment based on a color method is still continuously carried out, the detection technology using LAMP generally needs to keep warm for a period of time under isothermal conditions, so that amplification of target nucleic acid fragments is completed, rapid, efficient and specific nucleic acid detection is realized by observing the color change of a sample, cresol red is a red reagent sensitive to pH change, and as the pH value is lowered in the nucleic acid amplification process, the cresol red indicator changes from red to yellow, and can be used for naked eye detection of target pathogens. The color method based on the cresol red pH indicator for detecting isothermal amplification products has the advantages of simple system and stable reaction, and can be applied to the field of point of care testing (POCT), such as specific detection of campylobacter coli, campylobacter and salmonella. However, the method has the defects that the color is greatly affected by temperature, in the conventional LAMP detection for the color method based on cresol red, the detection result is usually determined by observing the color change of a sample by naked eyes after the sample is heated, the accuracy of the detection result is low, the detection quantity of detection equipment is small, high-flux detection cannot be realized, the detection result is finally obtained by different equipment, the detection equipment has a complex structure, the detection process is complex, the detection efficiency is low, and large-scale application cannot be realized.
Disclosure of Invention
Aiming at the problems of the background technology, the invention aims to provide a high-flux isothermal amplification one-stop detection device which can realize high-flux one-stop detection of a nucleic acid isothermal amplification reaction process, a sample shooting process and a detection result analysis process of LAMP detection, has simple structure, small volume and high detection efficiency, effectively improves the detection efficiency by real-time analysis, ensures the accuracy of the detection result, and solves the problems of low detection result accuracy, less detection quantity and low detection efficiency in the traditional color method LAMP detection based on cresol red;
The invention further aims to provide a use method of the high-flux isothermal amplification one-stop detection device, which can realize high-flux instant detection in a short time, is convenient to operate in a detection process and rapid in detection, and solves the problems of low LAMP detection efficiency, complex detection process and low detection efficiency of the traditional cresol red-based color method.
To achieve the purpose, the invention adopts the following technical scheme:
A high-throughput isothermal amplification one-stop detection device comprises a detection cassette, camera equipment, a cloud storage system and a computer system;
the detection cassette comprises a cassette body, a first placing surface is arranged on one side of the cassette body, a second placing surface is arranged on the side, opposite to the first placing surface, of the cassette body, the first placing surface and the second placing surface are obliquely arranged, a first support is arranged on the first placing surface and is used for supporting and placing a sample heating frame, a second support is arranged on the second placing surface and is used for supporting and placing the camera equipment;
The sample heating frame is provided with a plurality of sample holes for placing sample tubes, the sample holes penetrate through the sample heating frame, the first placing surface is provided with a through hole, the sample heating frame is placed at the position of the first placing surface corresponding to the through hole, the side surface of the sample heating frame is provided with a plurality of heating plates, and the second placing surface is provided with a shooting hole;
The bottom of the box body is in an open shape, a partition plate is horizontally arranged at the bottom of the box body, a mirror is arranged on the upper surface of the partition plate, and an included angle between the surface where the first placing surface is located and the surface where the second placing surface is located is 90 degrees;
the camera equipment is in signal connection with the cloud storage system, the cloud storage system is in signal connection with the computer system, and the camera equipment is used for photographing the sample color of the sample heating rack reflected by the mirror and sending the obtained sample detection image to the cloud storage system; and the computer system downloads the sample detection image from the cloud storage system for real-time analysis and judgment, and uploads the qualitative detection result to the cloud storage system.
Further, the surface of the box body is black, and the bottom surface of the sample heating rack is white.
Still further stated, the detection cassette further comprises a temperature controller and a thermocouple temperature sensor, the sample heating rack is connected with the input end of the thermocouple temperature sensor, the output end of the thermocouple temperature sensor is connected with the input end of the temperature controller, and the output end of the temperature controller is connected with the heating plate.
Further stated, the sample heating rack is provided with 64 sample holes, and the 64 sample holes are uniformly arranged on the sample heating rack, and the shape of the sample holes is matched with the shape of the tube body of the eight-connecting tube.
The application method of the high-throughput isothermal amplification one-stop detection device is applied to the high-throughput isothermal amplification one-stop detection device and comprises the following steps:
S1, adding a nucleic acid sample subjected to nucleic acid extraction into an LAMP reaction system of cresol red, uniformly mixing to obtain a sample to be detected, adding the sample to be detected into an octal pipe, placing the octal pipe into a sample hole of a sample heating frame for heating and carrying out isothermal amplification reaction, and after the reaction is finished, turning off a power supply of a heating plate, and cooling the temperature of the sample heating frame to room temperature;
s2, photographing through a photographing hole by using the photographing equipment, acquiring a sample detection image and sending the sample detection image to a cloud storage system;
And S3, the computer system downloads the sample detection image from the cloud storage system, performs real-time analysis and judgment, generates a qualitative detection result and then uploads the qualitative detection result to the cloud storage system.
Further describing, in the step S3, the sample detection image is analyzed and judged in real time, which specifically includes the following steps:
s31, dividing each sample detection hole to obtain the center coordinates of each sample detection hole;
S32, performing RGB color gamut conversion on the sample detection image to HSI color gamut;
s33, taking the average value of Hue channel values of all points as a detection value in the radius range of the center coordinates of each sample detection hole acquired in the step S31;
S34, counting the detection value of each sample detection hole, comparing the detection value of each sample detection hole with a threshold value, setting the detection result of the sample detection hole with the detection value larger than the threshold value as positive, setting the detection result of the sample detection hole with the detection value smaller than the threshold value as negative, and generating a qualitative detection result.
Further, in the step S31, the step of dividing each sample detection well specifically includes the steps of:
S311, filtering RGB channels of the sample detection image: setting a pixel point of which the R channel value is less than 100 to 0, setting a pixel point of which the G channel value is greater than 254 to 0, and setting a pixel point of which the B channel value is greater than 254 to 0;
S312, negating the sample detection image, and multiplying the negated image with the original image;
In step S31, the center coordinates of each sample detection hole are obtained, and the center coordinates are automatically detected by using a hough transform method, or manually selected by using a ginput () function of Matlab.
Further, in the step S33, the radius ranges from 4 to 6 pixels.
Further describing, in the step S32, the HSI color gamut is subjected to a process of rotating by 70 ° counterclockwise, and the specific rotation steps are as follows: the calculation is performed using the Hue value normalized by the original HSI gamut model, with h+0.194 > 1, then H-0.806, and h+0.194 <1, then h+0.194.
Further, in the step S34, the standard deviation of the detection value obtained by subtracting 5 times from the detection value obtained by reacting the plasmid with the lowest copy number in the cresol red system for 60min is used as the threshold.
Compared with the prior art, the embodiment of the invention has the following beneficial effects:
The high-flux isothermal amplification one-stop detection device can realize one-stop detection of a nucleic acid isothermal amplification reaction process, a sample shooting process and a detection result analysis process of LAMP detection, can realize high-flux detection of a plurality of samples, has a simple structure and a small volume, is convenient to assemble during detection, is suitable for large-scale application, effectively improves detection efficiency through real-time analysis, ensures accuracy of detection results, and can be applied to LAMP color method detection experiments using cresol red as an indicator.
Furthermore, the method for using the high-throughput isothermal amplification one-stop detection device is provided, the sample detection images are heated and shot by combining the sample detection images into the same device, the shooting equipment can send the sample detection images to the cloud storage system after shooting is completed, the computer system can download the sample detection images from the cloud storage system and conduct real-time analysis and judgment, the rapid detection is effectively realized, the whole detection time can be shortened to be within one hour, the analysis and judgment of the sample detection images can be completed within 5 minutes, the high-throughput instant detection can be realized in a short time, the detection process is convenient to operate, and the detection is rapid.
Drawings
The present invention is further illustrated by the accompanying drawings, which are not to be construed as limiting the invention in any way.
FIG. 1 is a schematic diagram of a high throughput isothermal amplification one-stop detection device according to one embodiment of the present invention (the cloud storage system and the computer system are not shown);
FIG. 2 is a schematic perspective view of a cartridge of a high throughput isothermal amplification one-stop detection device according to one embodiment of the present invention;
FIG. 3 is a schematic perspective view of a spacer plate of a high throughput isothermal amplification one-stop detection device according to one embodiment of the present invention;
FIG. 4 is a schematic diagram of S31 in a method of using a high throughput isothermal amplification one-stop detection device according to an embodiment of the present invention;
FIG. 5 is a schematic diagram of S32 in a method of using a high throughput isothermal amplification one-stop detection device according to an embodiment of the present invention;
FIG. 6 is a schematic diagram of S33 in a method of using a high throughput isothermal amplification one-stop detection device according to an embodiment of the present invention;
FIG. 7 is a schematic diagram of S34 in a method of using a high throughput isothermal amplification one-stop detection device according to an embodiment of the present invention;
FIG. 8 is a schematic representation of the detection results of campylobacter coli detection using a high throughput isothermal amplification one-stop detection device according to one embodiment of the present invention;
FIG. 9 is a schematic representation of the detection results of campylobacter coli detection using a high throughput isothermal amplification one-stop detection device according to one embodiment of the present invention;
wherein: the detection cassette 100, the cassette body 1, the first placement surface 11, the first holder 111, the through hole 112, the second wire via 113, the second placement surface 12, the second holder 121, the photographing hole 122, the sample heating rack 2, the sample hole 21, the image pickup apparatus 200, the heating sheet 4, the partition 5, the mirror 51, the first wire via 52, and the power supply box 6.
Detailed Description
Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative only and are not to be construed as limiting the invention.
In the description of the present invention, it should be understood that the orientation or positional relationship indicated by the terms "top", "bottom", "inner", etc. are based on the orientation or positional relationship shown in the drawings, and are merely for convenience of description and simplification of the description, and are not indicative or implying that the apparatus or element in question must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be construed as limiting the present invention. Furthermore, features defining "first", "second" may include one or more such features, either explicitly or implicitly, for distinguishing between the descriptive features, and not sequentially, and not lightly.
In the description of the present invention, unless otherwise indicated, the meaning of "a plurality" is two or more.
As shown in fig. 1 to 3, a high-throughput isothermal amplification one-stop detection device includes a detection cassette 100, a camera device 200, a cloud storage system, and a computer system;
The detection cassette 100 comprises a cassette body 1, a first placing surface 11 is arranged on one side of the cassette body 1, a second placing surface 12 is arranged on the side, opposite to the first placing surface 11, of the cassette body 1, the first placing surface 11 and the second placing surface 12 are obliquely arranged, the first placing surface 11 is provided with a first support 111, the first support 111 is used for supporting and placing the sample heating rack 2, the second placing surface 12 is provided with a second support 121, and the second support 121 is used for supporting and placing the image pickup device 200;
The sample heating rack 2 is provided with a plurality of sample holes 21 for placing sample tubes, the sample holes 21 penetrate through the sample heating rack 2, the first placing surface 11 is provided with a through hole 112, the sample heating rack 2 is placed at a position of the first placing surface 11 corresponding to the through hole 112, the side surface of the sample heating rack 2 is provided with a plurality of heating plates 4, and the second placing surface 12 is provided with a shooting hole 122;
The bottom of the box body 1 is in an open shape, a partition board 5 is horizontally arranged at the bottom of the box body 1, a mirror 51 is arranged on the upper surface of the partition board 5, and an included angle between the surface where the first placing surface 11 is located and the surface where the second placing surface 12 is located is 90 degrees;
The camera device 200 is in signal connection with the cloud storage system, the cloud storage system is in signal connection with the computer system, and the camera device 200 is used for photographing the sample color of the sample heating rack 2 reflected by the mirror 51 and sending the obtained sample detection image to the cloud storage system; and the computer system downloads the sample detection image from the cloud storage system for real-time analysis and judgment, and uploads the qualitative detection result to the cloud storage system.
Through setting up sample heating frame 2, can place the sample in sample hole 21 carries out the isothermal amplification reaction of nucleic acid in the in-process of heating, stop heating after reacting for a period, after the temperature drops to room temperature, use camera equipment 200 follow shooting hole 122 is shot the colour of sample, because sample hole 21 runs through sample heating frame 2, the surface of baffle 5 is equipped with mirror 51, can be with the sample colour reflection of natural light irradiation sample heating frame 2 to the shooting hole 122 of second place face 12, through with camera of camera equipment 200 aims at shooting hole 122, can acquire clear sample colour image, camera equipment 200 (camera equipment 200 specifically can adopt the smart mobile phone) can be with acquire sample detection image send to cloud memory system, computer system follow sample detection image is downloaded to the cloud memory system and is carried out real-time analysis and judge to sample detection image, finally generates and passes to after the testing result and carries out to memory system, camera equipment can follow the memory system download the testing result.
The included angle between the surface where the first placing surface 11 is located and the surface where the second placing surface 12 is located is 90 degrees, so that sample detection images of all samples can be completely shot, the volume of the detection device is effectively reduced by integrating heating of the samples and color shooting of the samples in one device, and when the detection device is used, a sample solution is located at the bottom of the sample hole 21, and color change of the samples can be observed at any time through the image pickup equipment 200.
The high-flux isothermal amplification one-stop detection device can realize one-stop detection of a nucleic acid isothermal amplification reaction process, a sample shooting process and a detection result analysis process of LAMP detection, can realize high-flux detection of a plurality of samples, has a simple structure and a small volume, is convenient to assemble during detection, is suitable for large-scale application, effectively improves detection efficiency through real-time analysis, ensures accuracy of detection results, and can be applied to LAMP color method detection experiments using cresol red as an indicator.
Preferably, the angle between the surface of the first placement surface 11 and the horizontal plane is 45 °, and the angle between the surface of the second placement surface 12 and the horizontal plane is 45 °.
Through setting up the contained angle of face that face 11 was located is 45 with the horizontal plane, the contained angle of face that face 12 was located is 45 with the horizontal plane to the second, guarantees the sample cell is placing in when the sample hole 21, the inclination of sample cell can not be too big, guarantees the stability of the isothermal amplification reaction process of nucleic acid, in addition, the inclination of face 12 is placed to the second also can not be too big, has guaranteed the stability of camera equipment 200 when shooing.
Specifically, two heating plates 4 are provided, two heating plates 4 are respectively provided on two sides of the sample heating frame 2, or three heating plates 4 are provided on three sides of the sample heating frame 2, and three heating plates 4 are respectively provided on three sides of the sample heating frame 2.
In general, the provision of two heating plates 4 can already satisfy the isothermal amplification reaction at about 62℃and, when the isothermal amplification reaction at 65℃or higher is required, the required heating temperature can be achieved by adding one of the heating plates 4, ensuring that the temperature requirement of the isothermal amplification reaction of nucleic acids can be satisfied.
Further, a power box 6 is further installed at the bottom of the partition board 5, and a transformer is arranged in the power box 6 and is electrically connected with the heating plates 4 through a plurality of power lines.
Specifically, the transformer is a 220V-to-12V transformer, and can stably output 10-12V adjustable voltage.
By setting the power supply box 6, the heating power supply voltage is output for the heating plate 4, so that the heating stability of the heating plate 4 is ensured, the heating effect of the heating plate 4 on the sample heating frame 2 is improved, and the heating effect of the nucleic acid isothermal amplification reaction process is ensured.
Further, the partition 5 is provided with a plurality of first wire passing holes 52 for the power wires to pass through, and the first wire passing holes 52 are arranged at one side of the partition 5 near the first placing surface 11 of the box 1;
the first placement surface 11 of the box body 1 is provided with a plurality of second wire through holes 113 for the power wires to pass through, and the second wire through holes 113 are arranged at positions, close to corners, of the first placement surface 11.
Specifically, six first wire through holes 52 are provided on a side of the partition board 5 near the first placement surface 11 of the box body 1, and four second wire through holes 113 are provided on the first placement surface 11 of the box body 1.
By providing the first via hole 52 and the second via hole 113, the power line may pass through the first via hole 52 and the second via hole 113, so as to ensure the stability of the heating operation of the heating plate 4.
Preferably, the sample heating rack 2 is made of aluminum material.
The sample heating frame 2 is made of aluminum materials, the aluminum materials have the advantages of light weight and good heat conductivity, the heat transfer efficiency between the heating plate 4 and the sample heating frame 2 in the isothermal amplification process and the heat transfer efficiency between the sample heating frame 2 and the sample tube can be ensured, and the heating effect is ensured.
Further, the surface of the case 1 is black, and the bottom surface of the sample heating rack 2 is white.
Specifically, the box body 1 is made of a conventional high-strength resin material, and the surface of the box body 1 is treated to be black by frosted paint.
The first placing surface 11 is provided with the through hole 112, the sample heating rack 2 is placed at a position of the first placing surface 11 corresponding to the through hole 112, and as the sample heating rack 2 is attached to the box body 1 when placed, the color of the surface of the box body 1 is black, so that light can be ensured to enter from the sample tube placed in the sample heating rack 2 only, in addition, the interference of stray light on the experimental result of a color method can be avoided during shooting, the deviation of the detection caused by the stray light is prevented, the external interference is effectively eliminated, the dark environment is provided for color observation, and the accuracy of the detection is ensured;
The color of the bottom surface of the sample heating rack 2 is white, so that diffuse reflection can be prevented, the color of the sample reflected by the mirror 51 is ensured to be the color of the sample hole 21 of the sample heating rack 2, and the color definition and accuracy of the shot color image are ensured.
Further, the detection cartridge 100 further includes a temperature controller and a thermocouple temperature sensor, the sample heating rack 2 is connected to an input end of the thermocouple temperature sensor, an output end of the thermocouple temperature sensor is connected to an input end of the temperature controller, and an output end of the temperature controller is connected to the heating sheet 4.
Specifically, the temperature controller and the thermocouple temperature sensor are disposed on the top surface of the case 1.
Through setting up temperature controller with thermocouple temperature sensor, thermocouple temperature sensor can acquire in real time the temperature of sample heating frame 2, thereby through temperature sensor is right the heating temperature of heating plate 4 controls, realizes the control to the sample heating temperature of sample heating frame 2, guarantees the temperature stability of nucleic acid isothermal amplification reaction process, thereby guarantees the accuracy of testing result.
Specifically, the sample heating rack 2 is provided with 64 sample holes 21, and the 64 sample holes 21 are uniformly arranged in the sample heating rack 2, and the shape of the sample holes 21 is matched with the shape of the tube body of the eight-connecting tube.
The sample heating frame 2 is provided with 64 sample holes 21, can be matched with eight connecting pipes, and can be used for placing the eight connecting pipes in the sample holes 21, so that high-flux detection can be realized, image shooting is convenient, and the detection accuracy is ensured, and meanwhile, the detection efficiency is effectively improved.
The application method of the high-throughput isothermal amplification one-stop detection device is applied to the high-throughput isothermal amplification one-stop detection device and comprises the following steps:
S1, adding a nucleic acid sample extracted from nucleic acid into an LAMP reaction system of cresol red, uniformly mixing to obtain a sample to be detected, adding the sample to be detected into an octal pipe, placing the octal pipe into a sample hole 21 of a sample heating frame 2, heating and carrying out isothermal amplification reaction, and after the reaction is finished, turning off a power supply of a heating plate 4, and cooling the temperature of the sample heating frame 2 to room temperature;
S2, the camera equipment 200 shoots through the shooting hole 122, obtains a sample detection image and sends the sample detection image to a cloud storage system;
And S3, the computer system downloads the sample detection image from the cloud storage system, performs real-time analysis and judgment, generates a qualitative detection result and then uploads the qualitative detection result to the cloud storage system.
By combining the sample heating and shooting of the sample detection image into the same device, the image pickup device 200 can send the sample detection image to the cloud storage system after shooting is completed, the computer system can download the sample detection image from the cloud storage system and perform real-time analysis and judgment, and the whole detection flow can pass through the image pickup device 200 (mobile phone end)And the software performs information intercommunication. /(I)APP can take pictures and upload local pictures to the cloud storage system (/ >Connector), then the computer system can download the sample detection image just uploaded from the cloud storage system, perform real-time analysis and judgment under the program location directory, generate a detection result, upload the detection result to the cloud storage system for storage, and the camera device 200 (mobile phone end) can download and view at any time. The rapid detection is effectively realized, the whole detection time can be shortened to be within one hour, the analysis and judgment of the sample detection image can be completed within 5 minutes, the high-flux instant detection can be realized in a short time, the operation of the detection process is convenient, and the detection is rapid.
Further describing, in the step S3, the sample detection image is analyzed and judged in real time, which specifically includes the following steps:
s31, dividing each sample detection hole to obtain the center coordinates of each sample detection hole;
S32, performing RGB color gamut conversion on the sample detection image to HSI color gamut;
s33, taking the average value of Hue channel values of all points as a detection value in the radius range of the center coordinates of each sample detection hole acquired in the step S31;
S34, counting the detection value of each sample detection hole, comparing the detection value of each sample detection hole with a threshold value, setting the detection result of the sample detection hole with the detection value larger than the threshold value as positive, setting the detection result of the sample detection hole with the detection value smaller than the threshold value as negative, and generating a qualitative detection result.
As shown in fig. 4 to fig. 7 (where Positive indicates that the detection result is Negative, positive indicates that the detection result is Positive), by separating each sample detection hole first, the Hue channel matrix of the whole sample detection image can be obtained after the color gamut is converted, at this time, the center coordinates of each sample detection hole obtained in the step S31 are used to obtain the Hue values of all points in the radius range of the center coordinates, the average value of the Hue values of all points in the radius range of the center coordinates is taken as the detection value, the detection value can represent the Hue channel value of the sample detection hole, and by adopting the Hue channel method to analyze the detection result, the stability of the determination of the detection result of the Negative sample and the Positive sample is better, the accuracy of the detection result is higher, the analysis and the determination of the sample detection image can be completed within 5min, the detection efficiency is high, and the detection stability is good.
As an embodiment of the present invention, the detection of campylobacter coli is performed using the high-throughput isothermal amplification one-stop detection device, comprising the steps of:
S1, adding a nucleic acid sample extracted from nucleic acid into an LAMP reaction system of cresol red, uniformly mixing to obtain a sample to be detected, adding the sample to be detected into an octal pipe, placing the octal pipe into a sample hole 21 of a sample heating frame 2, adjusting the heating temperature to 64 ℃, heating at 64 ℃ and carrying out isothermal amplification reaction for 30-45 min, and after the reaction is finished, turning off a power supply of a heating plate 4, and cooling the temperature of the sample heating frame 2 to room temperature;
S2, photographing through a photographing hole 122 by using a photographing device 200 (specifically adopting a smart phone), obtaining a sample detection image and sending the sample detection image to a computer system;
And S3, the computer system analyzes and judges the sample detection image in real time to generate an icon with a sample detection hole and a qualitative detection result of negative or positive information.
The high-throughput isothermal amplification one-stop detection device is used for detecting the campylobacter coli, positive and negative samples are placed at intervals, the obtained detection result is shown in a graph 8 (wherein Sample number refers to the number of the Sample, hue value refers to the Hue value of the Sample), the lighter color in the graph is a positive result (yellow), the darker color is a negative result (red), and the accuracy of the detection result is high;
The high-throughput isothermal amplification one-stop detection device is used for detecting the campylobacter coli, samples with unknown positives or negatives are placed randomly, the obtained detection results are shown in a figure 9 (wherein Sample number refers to the number of the Sample, hue value refers to the Hue value of the Sample), the lighter color is a positive result (yellow), the darker color is a negative result (red), and the stability of the detection results is strong.
Further, after the sample to be detected is added into the eight-connecting pipe, paraffin oil is added into the eight-connecting pipe to seal the sample to be detected and prevent pollution.
Further, in the step S31, the step of dividing each sample detection well specifically includes the steps of:
S311, filtering RGB channels of the sample detection image: setting a pixel point of which the R channel value is less than 100 to 0, setting a pixel point of which the G channel value is greater than 254 to 0, and setting a pixel point of which the B channel value is greater than 254 to 0;
S312, negating the sample detection image, and multiplying the negated image with the original image;
In step S31, the center coordinates of each sample detection hole are obtained, and the center coordinates are automatically detected by using a hough transform method, or manually selected by using a ginput () function of Matlab.
By dividing each of the sample detection holes by filtering RGB channels of a sample detection image, namely, setting a pixel point having a red channel (R channel) value smaller than 100 to 0, setting a pixel point having a green channel (G channel) value larger than 254 to 0, and setting a pixel point having a blue channel (B channel) value larger than 254 to 0, negating the filtered image, multiplying the negated image with an original image, an image having only a sample hole can be obtained, specifically, in a binary image, black is 1, white is 0, after negating, an original black part is 0, a white part is 1, multiplying the negated image with the original image, an image at the sample detection hole is retained, and an image other than the sample detection hole is 0, each of the sample detection holes can be divided for analysis;
The circle center coordinates are automatically detected by adopting the Hough transformation method, or the ginput () function of Matlab is adopted to manually select the circle center coordinates, when the circle center coordinates cannot be detected by adopting the Hough transformation method, the circle center coordinates can be selected by adopting the manual method, so that the fault tolerance rate in the detection process is effectively improved, and the circle center coordinates of the sample detection hole cannot be detected.
Preferably, the method further comprises the step of performing a corrosion calculation on the original sample detection image between the step of dividing each sample detection hole and the step of obtaining the center coordinates of each sample detection hole.
When the sample detection holes are identified by segmentation, noise points may appear in the sample detection image, and through corrosion calculation, the noise points between the sample detection holes can be removed, specifically, a single image of the sample detection holes is obtained by a mask method, and other areas in the image are black.
Preferably, in the step S33, the radius range is 4 to 6 pixels.
When the sample detection image is shot, pixel deviation may occur, the radius range of the center coordinates of each sample detection hole is set to be 4-6 pixels, if the radius range is too large, the value of the edge of the sample detection hole may be taken, the detection result is affected, if the radius range is too small, the value universality of the average value of the Hue channel values is reduced, and the accuracy of the detection result is also affected, so that each sample detection hole takes the average value of the Hue channel values in the radius range of 4-6 pixels as the detection value, and the accuracy of the detection result is ensured.
Preferably, in the step S32, the HSI color gamut is subjected to a process of rotating by 70 ° counterclockwise, and the specific rotation steps are as follows: the calculation is performed using the Hue value normalized by the original HSI gamut model, with h+0.194 > 1, then H-0.806, and h+0.194 < 1, then h+0.194.
Since the zero point of the original HSI gamut model is red, the color method reaction process using cresol red as an indicator is changed from magenta to yellow, and a reddish sample may appear, so that the HSI gamut is subjected to a process of rotating 70 ° counterclockwise, and is calculated by using the Hue value normalized by the original HSI gamut model, if h+0.194 > 1, the Hue value is H-0.806, and if h+0.194 < 1, the Hue value is h+0.194. (0.194=pi/180×70/2 pi), so that the zero point of the HSI color gamut is changed into bluish magenta, and the accuracy of the detection result is effectively improved.
Specifically, in the step S34, the standard deviation of the detection value obtained by subtracting 5 times from the detection value obtained by reacting the plasmid with the lowest copy number in the cresol red system for 60 minutes is used as the threshold.
As can be seen from the paired sample T-test, there is a significant difference between the detected values of the positive and negative samples (t= -31.56, P <0.01, T and P values are statistics, P value represents the minimum level of significance that does not accept the original hypothesis, and can be directly compared to the selected level of significance.e. taking a 5% level of significance, if the P value is greater than 5%, the original hypothesis is accepted, otherwise the original hypothesis is not accepted, i.e. the T-test demonstrates that there is a significant difference between negative and positive samples using the present method). By recording the standard deviation of the detection value of the plasmid (detection limit) with the lowest copy number after 60min of reaction in the cresol red system minus 5 times of the detection value as a threshold value, when the detection of the sample is carried out and the detection result is judged, the detection result of the sample detection hole with the detection value larger than the threshold value is positive, and the detection result of the sample detection hole with the detection value smaller than the threshold value is negative, so that the stability of the qualitative detection result is stronger.
The technical principle of the present invention is described above in connection with the specific embodiments. The description is made for the purpose of illustrating the general principles of the invention and should not be taken in any way as limiting the scope of the invention. Other embodiments of the invention will be apparent to those skilled in the art from consideration of this specification without undue burden.
Claims (3)
1. The application method of the high-throughput isothermal amplification one-stop detection device is characterized in that the high-throughput isothermal amplification one-stop detection device comprises a detection cassette, a camera device, a cloud storage system and a computer system;
the detection cassette comprises a cassette body, a first placing surface is arranged on one side of the cassette body, a second placing surface is arranged on the side, opposite to the first placing surface, of the cassette body, the first placing surface and the second placing surface are obliquely arranged, a first support is arranged on the first placing surface and is used for supporting and placing a sample heating frame, a second support is arranged on the second placing surface and is used for supporting and placing the camera equipment;
The sample heating frame is provided with a plurality of sample holes for placing sample tubes, the sample holes penetrate through the sample heating frame, the first placing surface is provided with a through hole, the sample heating frame is placed at the position of the first placing surface corresponding to the through hole, the side surface of the sample heating frame is provided with a plurality of heating plates, and the second placing surface is provided with a shooting hole;
The bottom of the box body is in an open shape, a partition plate is horizontally arranged at the bottom of the box body, a mirror is arranged on the upper surface of the partition plate, and an included angle between the surface where the first placing surface is located and the surface where the second placing surface is located is 90 degrees;
The camera equipment is in signal connection with the cloud storage system, the cloud storage system is in signal connection with the computer system, and the camera equipment is used for photographing the sample color of the sample heating rack reflected by the mirror and sending the obtained sample detection image to the cloud storage system; the computer system downloads the sample detection image from the cloud storage system for real-time analysis and judgment, and uploads the qualitative detection result to the cloud storage system;
The detection cassette further comprises a temperature controller and a thermocouple temperature sensor, the sample heating frame is connected with the input end of the thermocouple temperature sensor, the output end of the thermocouple temperature sensor is connected with the input end of the temperature controller, and the output end of the temperature controller is connected with the heating plate;
The application method of the high-throughput isothermal amplification one-stop detection device comprises the following steps:
S1, adding a nucleic acid sample subjected to nucleic acid extraction into an LAMP reaction system of cresol red, uniformly mixing to obtain a sample to be detected, adding the sample to be detected into an octal pipe, placing the octal pipe into a sample hole of a sample heating frame for heating and carrying out isothermal amplification reaction, and after the reaction is finished, turning off a power supply of a heating plate, and cooling the temperature of the sample heating frame to room temperature;
s2, photographing through a photographing hole by using the photographing equipment, acquiring a sample detection image and sending the sample detection image to a cloud storage system;
s3, the computer system downloads the sample detection image from the cloud storage system, performs real-time analysis and judgment, generates a qualitative detection result and then uploads the qualitative detection result to the cloud storage system;
In the step S3, the sample detection image is analyzed and judged in real time, which specifically includes the following steps:
s31, dividing each sample detection hole to obtain the center coordinates of each sample detection hole;
S32, performing RGB color gamut conversion on the sample detection image to HSI color gamut;
s33, taking the average value of Hue channel values of all points as a detection value in the radius range of the center coordinates of each sample detection hole acquired in the step S31;
S34, counting the detection value of each sample detection hole, comparing the detection value of each sample detection hole with a threshold value, setting the detection result of the sample detection hole with the detection value larger than the threshold value as positive, setting the detection result of the sample detection hole with the detection value smaller than the threshold value as negative, and generating a qualitative detection result;
In the step S31, the step of dividing each sample detection hole specifically includes the steps of:
S311, filtering RGB channels of the sample detection image: setting a pixel point of which the R channel value is less than 100 to 0, setting a pixel point of which the G channel value is greater than 254 to 0, and setting a pixel point of which the B channel value is greater than 254 to 0;
S312, negating the sample detection image, and multiplying the negated image with the original image;
In the step S31, the center coordinates of each sample detection hole are obtained, and the center coordinates are automatically detected by adopting a hough transformation method, or manually selected by adopting a ginput () function of Matlab;
In the step S32, the HSI color gamut is subjected to a process of rotating 70 ° counterclockwise, and the specific rotation steps are as follows: calculating a Hue value normalized by using an original HSI color gamut model, wherein if H+0.194 is more than 1, the Hue value is H-0.806, and if H+0.194 is less than 1, the Hue value is H+0.194;
In the step S33, the radius range is 4-6 pixel points;
in the step S34, the standard deviation of the detection value obtained by subtracting 5 times from the detection value obtained by reacting the plasmid with the lowest copy number in the cresol red system for 60min is used as the threshold.
2. The method of claim 1, wherein the surface of the cartridge is black and the bottom surface of the sample heating rack is white.
3. The method of claim 1, wherein 64 sample holes are formed in the sample heating rack, the 64 sample holes are uniformly arranged in the sample heating rack, and the shape of the sample holes is matched with the shape of the tube body of the eight-connecting tube.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110490627.XA CN113214974B (en) | 2021-05-06 | 2021-05-06 | High-flux isothermal amplification one-stop detection device and use method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110490627.XA CN113214974B (en) | 2021-05-06 | 2021-05-06 | High-flux isothermal amplification one-stop detection device and use method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113214974A CN113214974A (en) | 2021-08-06 |
| CN113214974B true CN113214974B (en) | 2024-05-17 |
Family
ID=77091418
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110490627.XA Active CN113214974B (en) | 2021-05-06 | 2021-05-06 | High-flux isothermal amplification one-stop detection device and use method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113214974B (en) |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6346384B1 (en) * | 2000-03-27 | 2002-02-12 | Dade Behring Inc. | Real-time monitoring of PCR using LOCI |
| WO2014058064A1 (en) * | 2012-10-12 | 2014-04-17 | 学校法人日本大学 | Inspection system and inspection method |
| CN109724972A (en) * | 2019-02-25 | 2019-05-07 | 深圳市象形字科技股份有限公司 | A kind of portable intelligent urine detection instrument and detection method |
| CN110734965A (en) * | 2019-11-06 | 2020-01-31 | 上海千履基因科技有限公司 | Method and device for quickly obtaining PCR (polymerase chain reaction) result of POCT (point of care testing) |
| WO2020089399A1 (en) * | 2018-10-31 | 2020-05-07 | Foundation For Research And Technology Hellas | Method and apparatus for performing a real-time colorimetric nucleic acid amplification assay |
| CN111197003A (en) * | 2019-12-26 | 2020-05-26 | 中国科学院合肥物质科学研究院 | Analysis device and method for integrated nucleic acid extraction amplification detection based on smart phone |
| CN111808744A (en) * | 2020-08-03 | 2020-10-23 | 齐鲁工业大学 | Portable full-automatic nucleic acid constant temperature amplification detector |
| CN111944678A (en) * | 2019-05-15 | 2020-11-17 | 青岛简码基因科技有限公司 | Portable nucleic acid colorimetric detection device and use method thereof |
| WO2020253464A1 (en) * | 2019-06-20 | 2020-12-24 | 北京科技大学 | Nucleic acid quantitative analysis method based on intelligent device assistance and applications thereof |
| CN112501010A (en) * | 2020-11-02 | 2021-03-16 | 济南大学 | Atmospheric pressure controlled on-chip reagent storage and delivery integrated nucleic acid detection device |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2006261346A1 (en) * | 2005-06-15 | 2006-12-28 | Gen-Probe Incorporated | Methods for quantitative analysis of a nucleic acid amplification reaction |
| US20140273181A1 (en) * | 2013-03-15 | 2014-09-18 | Biofire Diagnostics, Inc. | Compact optical system for substantially simultaneous monitoring of samples in a sample array |
| US20200063197A1 (en) * | 2017-05-15 | 2020-02-27 | Arizona Board Of Regents On Behalf Of Arizona State University | Quantitative detection and analysis of target dna with colorimetric rt-qlamp |
| US20210107008A1 (en) * | 2019-10-09 | 2021-04-15 | Genalyze LLC | System and method of isothermal DNA amplification devices with mobile devices for the cloud service |
-
2021
- 2021-05-06 CN CN202110490627.XA patent/CN113214974B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6346384B1 (en) * | 2000-03-27 | 2002-02-12 | Dade Behring Inc. | Real-time monitoring of PCR using LOCI |
| WO2014058064A1 (en) * | 2012-10-12 | 2014-04-17 | 学校法人日本大学 | Inspection system and inspection method |
| WO2020089399A1 (en) * | 2018-10-31 | 2020-05-07 | Foundation For Research And Technology Hellas | Method and apparatus for performing a real-time colorimetric nucleic acid amplification assay |
| CN109724972A (en) * | 2019-02-25 | 2019-05-07 | 深圳市象形字科技股份有限公司 | A kind of portable intelligent urine detection instrument and detection method |
| CN111944678A (en) * | 2019-05-15 | 2020-11-17 | 青岛简码基因科技有限公司 | Portable nucleic acid colorimetric detection device and use method thereof |
| WO2020253464A1 (en) * | 2019-06-20 | 2020-12-24 | 北京科技大学 | Nucleic acid quantitative analysis method based on intelligent device assistance and applications thereof |
| CN110734965A (en) * | 2019-11-06 | 2020-01-31 | 上海千履基因科技有限公司 | Method and device for quickly obtaining PCR (polymerase chain reaction) result of POCT (point of care testing) |
| CN111197003A (en) * | 2019-12-26 | 2020-05-26 | 中国科学院合肥物质科学研究院 | Analysis device and method for integrated nucleic acid extraction amplification detection based on smart phone |
| CN111808744A (en) * | 2020-08-03 | 2020-10-23 | 齐鲁工业大学 | Portable full-automatic nucleic acid constant temperature amplification detector |
| CN112501010A (en) * | 2020-11-02 | 2021-03-16 | 济南大学 | Atmospheric pressure controlled on-chip reagent storage and delivery integrated nucleic acid detection device |
Non-Patent Citations (2)
| Title |
|---|
| Competitive activation cross amplification combined with smartphone-based quantification for point-of-care detection of single nucleotide polymorphism;Junping Wen et al;《Biosensors and Bioelectronics》;第第183卷卷;第1-8页 * |
| 金属络合剂酸性络蓝K在TNOS-LAMP可视化检测体系中的应用;汪小福;付振芳;李莹;陈笑芸;彭城;徐晓丽;魏巍;徐俊锋;;农业生物技术学报(第09期);第1374-1383页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113214974A (en) | 2021-08-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ra et al. | Smartphone-based point-of-care urinalysis under variable illumination | |
| CN107680129B (en) | Portable bacterial colony automatic counting method based on smart phone | |
| CN115825070A (en) | Battery cell detection method and device, computer equipment and medium | |
| WO2019013960A1 (en) | Methods and systems for learning-based image edge enhancement of sample tube top circles | |
| CN113433120B (en) | Escherichia coli concentration detection method and system | |
| CN113214974B (en) | High-flux isothermal amplification one-stop detection device and use method | |
| CN109724398B (en) | Wood drying control method and device based on artificial intelligence | |
| BR112020025217A2 (en) | METHOD (110) TO EVALUATE AN ADEQUACY OF THE LIGHTING CONDITIONS, DETECTION METHOD TO DETECT AN ANALYTIS, COMPUTER PROGRAMS AND MOBILE DEVICE | |
| CN111079831A (en) | Intelligent optical detection sample characteristic and flaw automatic marking method and device | |
| CN103969193A (en) | sperm quality detection device | |
| CN110893399A (en) | Intelligent tobacco leaf grading and sorting equipment and method based on visual identification | |
| WO2022267799A1 (en) | Water quality testing method and water quality testing apparatus | |
| CN114169403A (en) | Coagulating picture shooting device, shooting method and flocculation control dosing method | |
| CN210916046U (en) | Real-time fluorescent quantitative PCR rapid detection equipment for remote diagnosis monitoring platform system | |
| CN111222360B (en) | Method, equipment and storage medium for detecting molten state of silicon material | |
| CN112950571B (en) | Method, device, equipment and computer storage medium for establishing yin-yang classification model | |
| CN109444207A (en) | A kind of melting point compound detector and its measuring method | |
| CN109919054B (en) | Machine vision-based reagent card automatic classification detection method | |
| CN111944678A (en) | Portable nucleic acid colorimetric detection device and use method thereof | |
| CN106790900A (en) | A kind of mobile phone temp detection method and system | |
| CN208636203U (en) | Portable milk protein content detection device | |
| CN111316934A (en) | Bee birth and fertility analyzer and method | |
| CN105917227A (en) | Quantitative real-time and end-point colorimetric PCR device | |
| CN214173650U (en) | Lees automatic feeding device based on intelligence vision | |
| CN211522208U (en) | Real-time digital RGB color imaging quantitative PCR instrument |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |