CN114381363A - Preparation method of PCR (polymerase chain reaction) rapid detection system and PCR rapid detection system - Google Patents
Preparation method of PCR (polymerase chain reaction) rapid detection system and PCR rapid detection system Download PDFInfo
- Publication number
- CN114381363A CN114381363A CN202111629112.XA CN202111629112A CN114381363A CN 114381363 A CN114381363 A CN 114381363A CN 202111629112 A CN202111629112 A CN 202111629112A CN 114381363 A CN114381363 A CN 114381363A
- Authority
- CN
- China
- Prior art keywords
- detection system
- preparing
- chip
- rapid detection
- pcr
- 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.)
- Granted
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 95
- 238000002360 preparation method Methods 0.000 title claims abstract description 29
- 238000003752 polymerase chain reaction Methods 0.000 title abstract description 97
- 239000002184 metal Substances 0.000 claims abstract description 54
- 229910052751 metal Inorganic materials 0.000 claims abstract description 54
- 238000000018 DNA microarray Methods 0.000 claims abstract description 44
- 238000000151 deposition Methods 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims description 47
- 238000004140 cleaning Methods 0.000 claims description 14
- 230000035945 sensitivity Effects 0.000 claims description 13
- 238000004458 analytical method Methods 0.000 claims description 11
- 229920002120 photoresistant polymer Polymers 0.000 claims description 11
- 238000004088 simulation Methods 0.000 claims description 11
- 238000013461 design Methods 0.000 claims description 10
- 238000005530 etching Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 7
- 229910052737 gold Inorganic materials 0.000 claims description 7
- 230000007480 spreading Effects 0.000 claims description 7
- 238000003892 spreading Methods 0.000 claims description 7
- 229910052782 aluminium Inorganic materials 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 229910052709 silver Inorganic materials 0.000 claims description 4
- 238000002944 PCR assay Methods 0.000 claims 2
- 238000010438 heat treatment Methods 0.000 abstract description 6
- 239000004065 semiconductor Substances 0.000 abstract description 3
- 230000008569 process Effects 0.000 description 18
- 206010028980 Neoplasm Diseases 0.000 description 13
- 201000011510 cancer Diseases 0.000 description 12
- 238000004519 manufacturing process Methods 0.000 description 7
- 238000012216 screening Methods 0.000 description 7
- 238000012795 verification Methods 0.000 description 7
- 239000004020 conductor Substances 0.000 description 6
- 230000005669 field effect Effects 0.000 description 6
- 238000004969 ion scattering spectroscopy Methods 0.000 description 6
- 238000004020 luminiscence type Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 238000012031 short term test Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005382 thermal cycling Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 206010035664 Pneumonia Diseases 0.000 description 2
- 229910052681 coesite Inorganic materials 0.000 description 2
- 229910052906 cristobalite Inorganic materials 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000005674 electromagnetic induction Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000000377 silicon dioxide Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 229910052682 stishovite Inorganic materials 0.000 description 2
- 229910052905 tridymite Inorganic materials 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001312 dry etching Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000003014 reinforcing effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 229910001868 water Inorganic materials 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
-
- 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
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Clinical Laboratory Science (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a preparation method of a PCR (polymerase chain reaction) rapid detection system and the PCR rapid detection system, and relates to the technical field of semiconductors and bioengineering. The preparation method of the PCR rapid detection system comprises the following steps: providing or preparing a biochip; depositing a metal layer on the surface of the biochip; providing or preparing a light emitting chip having a light emitting array; providing or preparing a driving chip electrically connected with the light-emitting chip; bonding the light-emitting chip and the driving chip, and bonding one side of the biochip provided with the metal layer to the light-emitting chip to obtain the PCR rapid detection system; the light emitted by the light emitting chip can irradiate the metal layer, so that the temperature of the metal layer is changed. The invention solves the technical problem of lower heating efficiency of the PCR rapid detection system in the prior art.
Description
Technical Field
The invention relates to the technical field of semiconductors and bioengineering, in particular to a preparation method of a PCR (polymerase chain reaction) rapid detection system and the PCR rapid detection system.
Background
Current DNA detection techniques based on PCR (optical polymerase chain reaction) require tens of thermal cycles to complete the detection. The traditional PCR reaction scheme based on a micro-heater has low temperature change speed, is a main reason for long time consumption of the thermal cycle of the whole DNA detection system, and causes low detection efficiency.
Disclosure of Invention
In view of this, the invention provides a preparation method of a rapid PCR detection system and a rapid PCR detection system, which are used for solving the technical problem that the rapid PCR detection system in the prior art consumes long time in thermal cycle.
In order to solve the technical problems, the first technical scheme adopted by the invention is as follows:
a preparation method of a PCR rapid detection system comprises the following steps:
providing or preparing a biochip;
depositing a metal layer on the surface of the biochip;
providing or preparing a light emitting chip having a light emitting array;
providing or preparing a driving chip electrically connected with the light-emitting chip; and
bonding the light-emitting chip and the driving chip, and bonding one side of the biochip provided with the metal layer to the light-emitting chip to obtain the PCR rapid detection system; the light emitted by the light emitting chip can irradiate the metal layer, so that the temperature of the metal layer is changed.
In some embodiments of the method for preparing a PCR rapid test system, the method for preparing a PCR rapid test system further comprises the steps of: and etching micropores on the metal layer.
In some embodiments of the method for preparing a PCR rapid test system, the method for preparing a PCR rapid test system further comprises the steps of:
obtaining parameters of an ion sensitive sensor and a temperature sensor in the biochip based on analysis simulation design; and
and integrating the ion sensitive sensor and the temperature sensor on the same carrier to form a sensor system.
In some embodiments of the method for preparing a PCR rapid detection system, the material of the metal layer includes any one of Au, Ag, and Al.
In some embodiments of the PCR rapid detection system preparation method, the sensitivity of the ion sensitive sensor is no less than 190mV/pH and the accuracy of the temperature sensor is no less than 0.2 ℃.
In some embodiments of the preparation method of the PCR rapid detection system, the light emitting chip is a Micro-LED chip, and the light wavelength range emitted by the light emitting chip is 400-850 nm.
In some embodiments of the method for preparing a PCR rapid detection system, the method for preparing the light emitting chip comprises the following steps:
providing or preparing an epitaxial wafer;
cleaning the epitaxial wafer;
drying the cleaned epitaxial wafer, and etching the table top;
removing the photoresist of the epitaxial wafer with the etched table top, and cleaning and drying;
forming a current spreading layer on the mesa; and
and evaporating an electrode on the current spreading layer.
In some embodiments of the method for manufacturing a PCR rapid detection system, the bonding mode of the light emitting chip and the driving chip is flip-chip bonding, and the bonding mode of the biochip and the light emitting chip is flip-chip bonding.
In some embodiments of the method for preparing a PCR rapid detection system, the number of the microwells is multiple, and each microwell is uniformly distributed on the metal layer.
In order to solve the technical problems, the invention adopts the following technical scheme:
a PCR rapid detection system is prepared by the preparation method of the PCR rapid detection system in the embodiment.
The embodiment of the invention has the following beneficial effects:
compared with the traditional mode of heating the matrix to be detected by adopting a traditional heater, the PCR rapid detection system obtained by the preparation method of the PCR rapid detection system has the advantages that the time required by the thermal cycle reaction is greatly reduced, and the technical problem of long time consumption of the thermal cycle of the PCR rapid detection system in the prior art is solved. Specifically, the preparation method of the PCR rapid detection system comprises the steps of providing or preparing a biochip; depositing a metal layer on the surface of the biochip; providing or preparing a light emitting chip having a light emitting array; providing or preparing a driving chip for electrically connecting with the light-emitting chip; and bonding the light-emitting chip and the driving chip, and bonding one side of the biochip, which is provided with the metal layer, to the light-emitting chip to obtain the PCR rapid detection system. Through the light that will send out the luminescence chip shine to the metal level on to can change the temperature of metal level, can quick adjustment metal level's temperature through the luminescence chip for the thermal cycle ability has been accelerated, thereby shortens the time of thermal cycle reaction. According to the invention, the light-emitting chip, the biochip and the driving chip are integrated to form a PCR rapid detection system, and the thermal cycle efficiency is improved in a mode of rapidly changing the temperature of the metal layer through the light-emitting chip, so that the technical problem that the PCR rapid detection system in the prior art is long in thermal cycle time consumption is solved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a PCR rapid detection system according to the present invention;
FIG. 2 is an exploded view of a part of the PCR rapid detection system of the present invention;
FIG. 3 is a graph of time versus temperature for thirty thermal cycles in accordance with the present invention;
FIG. 4 is a flow chart of the preparation method of the present invention.
Wherein: 1. a biochip; 2. a light emitting chip; 3. a driving chip; 4. a metal layer.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. The invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Current DNA detection techniques based on PCR (optical polymerase chain reaction) require tens of thermal cycles to complete the detection. The traditional PCR reaction scheme based on a micro-heater is slow in speed, is the main reason of long time consumption of the whole DNA detection system, and causes low detection efficiency.
Referring to fig. 1-2 and fig. 4, in an embodiment of a method for preparing a PCR rapid detection system, the method comprises the following steps:
providing or preparing a biochip 1;
depositing a metal layer 4 on the surface of the biochip 1;
providing or preparing a light emitting chip 2 having a light emitting array;
providing or preparing a driving chip 3 for electrically connecting with the light-emitting chip 2; and
the light-emitting chip 2 is bonded with the driving chip 3, and then the side of the biochip 1 provided with the metal layer 4 is bonded with the light-emitting chip 2 to obtain a PCR rapid detection system, and light emitted by the light-emitting chip 2 can irradiate the metal layer 4, so that the temperature of the metal layer 4 is changed.
In the preparation method of the PCR rapid detection system in this embodiment, the metal layer 4 deposited on the surface of the biochip 1 can receive the light from the light emitting chip 2, and the temperature of the metal layer 4 can be changed by adjusting the driving chip 3, so that the thermal cycling capability of the whole PCR rapid detection system can be accelerated, the time of thermal cycling reaction can be shortened, and the technical problem of long thermal cycling time consumption of the PCR rapid detection system in the prior art can be solved.
In an embodiment of the method for preparing a PCR rapid detection system, the method for preparing a PCR rapid detection system further comprises the following steps: micro-holes are etched in the metal layer 4.
In this embodiment, specifically, the micropore is located one side of metal level 4 orientation luminescence chip 2, can be the blind hole, also can be for the through-hole that runs through metal level 4, makes the surface area increase of metal level 4, when the light that luminescence chip 2 sent shines metal level 4, can increase with the area of contact of light, improves heat conduction efficiency, reduces the heat loss to reach the effect of reinforcing PCR rapid detection system's power consumption utilization ratio.
Preferably, the number of the micro holes is plural, and each micro hole is uniformly distributed on the metal layer 4. Specifically, the structure in which each fine pore constitutes a matrix is arranged on the metal layer 4.
In one embodiment of a method for preparing a PCR rapid detection system, the step of preparing the biochip 1 comprises:
obtaining parameters of an ion sensitive sensor and a temperature sensor in the biochip 1 based on analysis simulation design; and
the heater, the ion sensitive sensor and the temperature sensor are integrated on the same carrier to form a sensor system.
In this embodiment, specifically, the analysis and simulation adopts a gm/Id-based mode, and based on theoretical analysis, model simulation and experimental verification of gm/Id, parameters such as the optimal bias voltage, the working interval and the like of the isfet can be obtained, and the obtained parameters are used for design and development of the sensor system, so that the optimal performance can be obtained conveniently.
In addition, the carrier can be a wafer, and an interface circuit of the whole sensor system is also integrated on the same wafer, so that the sensitivity of the whole sensor system can be improved, and the accuracy and the reliability of the whole sensor system can be improved.
In an embodiment of the preparation method of the PCR rapid detection system, the material of the metal layer 4 comprises any one of Au, Ag and Al.
In the present invention, the metal layer 4 serves to receive light emitted from the light emitting chip 2 and thereby change the temperature of the metal layer 4 itself. Wherein preferred material is Au, and Au's heat conductivity is good, can improve heat-conducting efficiency, improves detection efficiency to can cooperate luminescence chip 2 to reach rapid heating up cooling and realize the effect of quick thermal cycle.
In an embodiment of the preparation method of the PCR rapid detection system, the preparation of the driving chip 3 is based on a CMOS process.
In this embodiment, the CMOS process is a short for complementary metal oxide semiconductor, which is developed based on PMOS and NMOS processes, and the integrated circuit based on the CMOS process has the advantages of low power consumption, high speed, strong anti-interference capability, and high integration level, and the preparation of the driving chip 3 and the sensor system of the present invention is based on the CMOS process, so that the cost of the PCR rapid detection system of the present invention can be reduced to a level that can be accepted by a large number of consumers, and the CMOS process itself has high maturity and low difficulty, thereby facilitating large-scale production and application of manufacturers, facilitating purchase and use of more consumers, facilitating use of consumers, facilitating early screening of diseases, and reducing middle and later stage cases.
In the embodiment of the preparation method of the PCR rapid detection system, the sensitivity of the ion sensitive sensor is not lower than 190mV/pH, and the accuracy of the temperature sensor is not lower than 0.2 ℃.
In the embodiment, the sensitivity of the ion sensitive sensor can determine the reading time of the signal, and when the sensitivity reaches more than 190mV/pH, microsecond-level signal reading can be achieved, so that the accuracy and the reliability of the whole sensor system are improved.
In an embodiment of a method for preparing a PCR rapid detection system, the light emitting chip 2 is a Micro-LED chip. The light wavelength range emitted by the light emitting chip 2 is 400nm-850 nm.
The Micro-LED is a light emitting chip 2 array formed by thinning and shrinking a traditional LED, and is generally an LED chip with the size less than 50 um. The Micro-LED has good monochromaticity, narrow half-wave width and high light efficiency, so that the structure for receiving light has higher heat receiving.
The size of the Micro-LED is nano-scale, and the photon PCR based on the Micro-LED can be prepared into smaller size, is not limited by the size of a light source, and has more application and development space.
The service life of the Micro-LED is ten times that of the common LED, the light source does not need to be frequently replaced, and the service life of the PCR can be prolonged.
Based on the excellent characteristics of GaN, the Micro-LED prepared from GaN can operate in extreme environments (such as strong acid, strong base, high and low temperature), is corrosion resistant, has good stability, and can operate in various environments.
The Micro-LED has low power consumption, the luminous efficiency is 6-7 times of that of the traditional light source, and the energy consumption is greatly reduced. In addition, no harmful substance is generated in the preparation process and the use process of the Micro-LED, so that the emission of pollutants and harmful substances is reduced, and the Micro-LED is an ideal clean light source. The Micro-LED light source is used for driving the PCR, and the experimental result is not influenced by chemical pollution.
In an embodiment of a method for preparing a PCR rapid detection system, the method for preparing the light emitting chip comprises the following steps:
providing or preparing an epitaxial wafer;
cleaning the epitaxial wafer;
drying the cleaned epitaxial wafer, and etching the table top;
removing the photoresist of the epitaxial wafer with the etched table top, and cleaning and drying;
forming a current spreading layer on the mesa; and
and evaporating an electrode on the current spreading layer.
Specifically, in this embodiment, the reagent for cleaning the epitaxial wafer may be in accordance with H2SO4：H2O2：H2O is 5: 1: 1, cleaning for 10min, cleaning with acetone for 5min, cleaning with isopropanol for 5min, cleaning with deionized water, and blowing with nitrogen.
The specific method for mesa etching comprises depositing 300nm SiO2And then dry etching is performed. The specific process parameters are as follows: over-etching 40% of SiO2The etching rate was 30nm/min, and the etching rate of the photoresist was 18A/min.
The photoresist removing mode is a plasma dry photoresist removing mode, and the specific technological parameters are as follows: 400W, O2The flow rate and time of (2) were 120sccm and 3min, respectively. After removing the photoresist, cleaning the photoresist by using acetone for 5min, then cleaning the photoresist by using isopropanol for 5min, finally cleaning the photoresist by using deionized water, and drying the photoresist by using nitrogen.
The current spreading layer is made of ITO or Ni or Au. The process of evaporating the electrode is an ELEC process.
According to the preparation method of the PCR rapid detection system, the luminescent chip 2, the driving chip 3 and the biochip 1 are connected in a process that the luminescent chip 2 and the driving chip 3 are bonded in an inverted mode, and then the biochip 1 and the luminescent chip 2 are bonded in an inverted mode. Through the mode of flip-chip bonding, can reduce the volume size of whole PCR short-term test system.
The invention also relates to a PCR rapid detection system, which is prepared by the preparation method of the PCR rapid detection system in the embodiment. When the PCR rapid detection system is applied to PCR equipment, the reaction vessel can be arranged on the PCR equipment and is used for accommodating a matrix to be detected.
Through being based on CMOS technology, can make the cost reduction of PCR short-term test system to reduce PCR short-term test system's cost, vast consumer can both receive, makes things convenient for the screening of early case, reduces middle and later stage severe. For example, early screening for cancer, cancer is still the leading cause of death worldwide, and its morbidity and mortality are still rising year by year. In China, the incidence of cancer is close to the world level, but the mortality rate is higher than the world level. According to the latest statistics of the current state of cancer, the total cancer rate of China reaches 5.5 per thousand, one person can be diagnosed as cancer every 6 minutes, 8550 people become cancer patients every day, and one person dies of cancer every seven to eight people. The cancer morbidity and mortality in china will continue to rise in the next 10 years. The mortality rate of cancer is high, and one important reason is that most cancers are found in middle and late stages in China. By adopting the PCR rapid detection system, the consumer can be reserved at home, so that the consumer can conveniently screen family members, early discovery is realized, the generation of middle and later period cases is reduced, and the loss of life and property can be reduced in the long run.
The invention can compress the whole detection time by heating the micro-heater and the light-emitting chip 2, as shown in figure 3, the thermal cycle time in the PCR rapid detection system can be compressed to be within 5 minutes, the time cost is reduced, and the rapid, accurate and on-site disease screening becomes more reliable.
In a further preferred embodiment, as shown in fig. 4, the method for preparing the PCR rapid detection system of the present invention comprises:
obtaining the optimal bias voltage and working interval parameters of the ion scattering field effect transistor based on the theoretical analysis, model simulation and experimental verification of gm/Id;
designing an ion sensitive sensor and a temperature sensor based on the obtained parameters, and integrating the ion sensitive sensor and the temperature sensor into a sensor system;
applying the obtained sensor system to the design and manufacture of the biochip 1 to obtain the biochip 1;
depositing a metal layer 4 made of any one of Au, Ag and Al on the obtained biochip 1;
preparing a Micro-LED chip, wherein the wavelength range of light emitted by the Micro-LED chip is 400nm to 850 nm;
preparing a driving chip 3 based on a CMOS process; and
the Micro-LED chip is in flip-chip bonding with the driving chip 3, the biological chip 1 is in flip-chip bonding with the Micro-LED chip, and the metal layer 4 faces to the Micro-LED chip;
wherein, the sensitivity of the ion sensitive sensor is not lower than 190mV/pH, and the accuracy of the temperature sensor is not lower than 0.2 ℃.
The first specific embodiment of the preparation method of the PCR rapid detection system of the invention is as follows:
obtaining the optimal bias voltage and working interval parameters of the ion scattering field effect transistor based on the theoretical analysis, model simulation and experimental verification of gm/Id;
designing an ion sensitive sensor and a temperature sensor based on the obtained parameters, and integrating the ion sensitive sensor and the temperature sensor into a sensor system;
applying the obtained sensor system to the design and manufacture of the biochip 1 to obtain the biochip 1;
depositing an Au metal layer 4 on the obtained biochip 1;
preparing a Micro-LED chip, wherein the wavelength of light emitted by the Micro-LED chip is 450 nm;
preparing a driving chip 3 based on a CMOS process; and
the Micro-LED chip is in flip-chip bonding with the driving chip 3, the biological chip 1 is in flip-chip bonding with the Micro-LED chip, and the metal layer 4 faces to the Micro-LED chip;
wherein the sensitivity of the ion sensitive sensor is 190mV/pH, and the accuracy of the temperature sensor is 0.2 ℃.
The PCR rapid detection system prepared by the above embodiment has a thermal cycle time of 250 seconds for thirty times.
The specific embodiment of the preparation method of the PCR rapid detection system of the invention is as follows:
obtaining the optimal bias voltage and working interval parameters of the ion scattering field effect transistor based on the theoretical analysis, model simulation and experimental verification of gm/Id;
designing an ion sensitive sensor and a temperature sensor based on the obtained parameters, and integrating the ion sensitive sensor and the temperature sensor into a sensor system;
applying the obtained sensor system to the design and manufacture of the biochip 1 to obtain the biochip 1;
depositing an Au metal layer 4 on the obtained biochip 1;
preparing a Micro-LED chip, wherein the wavelength of light emitted by the Micro-LED chip is 850 nm;
preparing a driving chip 3 based on a CMOS process; and
the Micro-LED chip is in flip-chip bonding with the driving chip 3, the biological chip 1 is in flip-chip bonding with the Micro-LED chip, and the metal layer 4 faces to the Micro-LED chip;
wherein the sensitivity of the ion sensitive sensor is 190mV/pH, and the accuracy of the temperature sensor is 0.2 ℃.
The PCR rapid detection system prepared by the above embodiment has a thermal cycle time of 270 seconds for thirty times.
The specific embodiment of the preparation method of the PCR rapid detection system of the invention is as follows:
obtaining the optimal bias voltage and working interval parameters of the ion scattering field effect transistor based on the theoretical analysis, model simulation and experimental verification of gm/Id;
designing an ion sensitive sensor and a temperature sensor based on the obtained parameters, and integrating the ion sensitive sensor and the temperature sensor into a sensor system;
applying the obtained sensor system to the design and manufacture of the biochip 1 to obtain the biochip 1;
depositing an Ag metal layer 4 on the obtained biochip 1;
preparing a Micro-LED chip, wherein the wavelength of light emitted by the Micro-LED chip is 650 nm;
preparing a driving chip 3 based on a CMOS process; and
the Micro-LED chip is in flip-chip bonding with the driving chip 3, the biological chip 1 is in flip-chip bonding with the Micro-LED chip, and the metal layer 4 faces to the Micro-LED chip;
wherein the sensitivity of the ion sensitive sensor is 200mV/pH, and the accuracy of the temperature sensor is 0.1 ℃.
The PCR rapid test system prepared by the above specific example has a thermal cycle time of 275 seconds for thirty times.
The third specific embodiment of the preparation method of the PCR rapid detection system of the invention:
obtaining the optimal bias voltage and working interval parameters of the ion scattering field effect transistor based on the theoretical analysis, model simulation and experimental verification of gm/Id;
designing an ion sensitive sensor and a temperature sensor based on the obtained parameters, and integrating the ion sensitive sensor and the temperature sensor into a sensor system;
applying the obtained sensor system to the design and manufacture of the biochip 1 to obtain the biochip 1;
depositing an Al metal layer 4 on the obtained biochip 1;
preparing a Micro-LED chip, wherein the wavelength of light emitted by the Micro-LED chip is 400 nm;
preparing a driving chip 3 based on a CMOS process; and
the Micro-LED chip is in flip-chip bonding with the driving chip 3, the biological chip 1 is in flip-chip bonding with the Micro-LED chip, and the metal layer 4 faces to the Micro-LED chip;
wherein the sensitivity of the ion sensitive sensor is 190mV/pH, and the accuracy of the temperature sensor is 0.2 ℃.
The PCR rapid detection system prepared by the above embodiment has a thermal cycle time of thirty times of 280 seconds.
The third specific embodiment of the preparation method of the PCR rapid detection system of the invention:
obtaining the optimal bias voltage and working interval parameters of the ion scattering field effect transistor based on the theoretical analysis, model simulation and experimental verification of gm/Id;
designing an ion sensitive sensor and a temperature sensor based on the obtained parameters, and integrating the ion sensitive sensor and the temperature sensor into a sensor system;
applying the obtained sensor system to the design and manufacture of the biochip 1 to obtain the biochip 1;
depositing an Al metal layer 4 on the obtained biochip 1;
preparing a Micro-LED chip, wherein the wavelength range of light emitted by the Micro-LED chip is 460 nm;
preparing a driving chip 3 based on a CMOS process; and
the Micro-LED chip is in flip-chip bonding with the driving chip 3, the biological chip 1 is in flip-chip bonding with the Micro-LED chip, and the metal layer 4 faces to the Micro-LED chip;
wherein the sensitivity of the ion sensitive sensor is not lower than 190 mV/the accuracy of the pH temperature sensor is not lower than 0.2 ℃.
The PCR rapid detection system prepared by the above embodiment has a thermal cycle time of 290 seconds for thirty times.
The above embodiments are only some embodiments of the present invention, and actually many embodiments of the specific preparation method are needed, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described. In these embodiments, the metal layer 4 is made of Au, and the light-emitting chip 2 has a wavelength of 450nm, which is the best effect when the two are used together. The improvement of the sensitivity of the ion sensitive sensor and the accuracy of the temperature sensor has little influence on the reduction of the thermal cycle time, the improvement cost is high, and the specific selection can be selected according to the actual situation.
As shown in FIG. 3, the PCR rapid detection system of the present invention has thirty thermal cycles (from 95 ℃ denaturation to 55 ℃ annealing and extension) of less than 5 minutes during the PCR reaction, and the extremely short PCR reaction time can greatly increase the response speed of the whole PCR rapid detection system, so that rapid, accurate and on-site disease screening becomes more reliable.
The PCR rapid detection system and the PCR rapid detection system can be used for early cancer detection, such as rapid epidemic screening of new coronary pneumonia and food hygiene detection, wherein a template of PCR reaction is DNA, so that the detection of RNA such as new coronary pneumonia and the like needs to be firstly reverse-transcribed into DNA and then tested. By the rapid PCR detection system and the rapid PCR detection system, the total reaction time is about 5 minutes, so that the rapid diagnosis of early cancer is realized, and the time of a user cannot be wasted due to long-time waiting. Patients and healthy people can be quickly screened in epidemic disease screening, so that epidemic situation defense is more scientific. And the food hygiene can improve the detection rate of the food and quickly screen problem food.
In addition, the existing PCR rapid detection system generally uses a micro-heater as a heat source, such as a resistance type micro-heater and an electromagnetic induction type micro-heater, the resistance type micro-heater is heated by using the resistance of a conductor to generate heat, current passes through the conductor, the conductor has a blocking effect on the current, the current overcomes the resistance of the conductor to do work and is transmitted through electric energy, the electric energy is converted into heat energy, and the heating is slow; the electromagnetic induction heater heats the conductor by adopting the heat effect generated by the induced current formed by the conductor in the alternating electromagnetic process, the heating speed is relatively slow, the detection time of thirty times of heat cycles is more than 40 minutes, and the time is long.
The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, but should be considered as the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above examples only show some embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the claims. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A preparation method of a PCR rapid detection system is characterized by comprising the following steps:
providing or preparing a biochip;
depositing a metal layer on the surface of the biochip;
providing or preparing a light emitting chip having a light emitting array;
providing or preparing a driving chip electrically connected with the light-emitting chip; and
bonding the light-emitting chip and the driving chip, and bonding one side of the biochip provided with the metal layer to the light-emitting chip to obtain a PCR rapid detection system; the light emitted by the light emitting chip can irradiate the metal layer, so that the temperature of the metal layer is changed.
2. The method for preparing a rapid PCR assay system of claim 1, further comprising the steps of: and etching micropores on the metal layer.
3. The method for preparing a rapid PCR assay system of claim 2, further comprising the steps of:
obtaining parameters of an ion sensitive sensor and a temperature sensor in the biochip based on analysis simulation design; and
and integrating the ion sensitive sensor and the temperature sensor on the same carrier to form a sensor system.
4. The method for preparing a PCR rapid detection system according to claim 3, wherein the material of the metal layer comprises any one of Au, Ag and Al.
5. The method for preparing a PCR rapid detection system according to claim 3, wherein the sensitivity of the ion sensitive sensor is not lower than 190mV/pH, and the accuracy of the temperature sensor is not lower than 0.2 ℃.
6. The method for preparing a PCR rapid detection system according to claim 1, wherein the light emitting chip is a Micro-LED chip, and the light emitted from the light emitting chip has a wavelength ranging from 400nm to 850 nm.
7. The method for preparing a PCR rapid detection system according to claim 6, wherein the method for preparing the light emitting chip comprises the following steps:
providing or preparing an epitaxial wafer;
cleaning the epitaxial wafer;
drying the cleaned epitaxial wafer, and etching the table top;
removing the photoresist of the epitaxial wafer with the etched table top, and cleaning and drying;
forming a current spreading layer on the mesa; and
and evaporating an electrode on the current spreading layer.
8. The method for preparing a PCR rapid detection system according to claim 1, wherein the bonding mode of the luminescent chip and the driver chip is flip-chip bonding, and the bonding mode of the biochip and the luminescent chip is flip-chip bonding.
9. The method of claim 2, wherein the number of the micro wells is multiple, and each micro well is uniformly distributed on the metal layer.
10. A PCR rapid detection system, which is prepared by the method for preparing a PCR rapid detection system according to any one of claims 1 to 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111629112.XA CN114381363B (en) | 2021-12-28 | 2021-12-28 | Preparation method of PCR rapid detection system and PCR rapid detection system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111629112.XA CN114381363B (en) | 2021-12-28 | 2021-12-28 | Preparation method of PCR rapid detection system and PCR rapid detection system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114381363A true CN114381363A (en) | 2022-04-22 |
| CN114381363B CN114381363B (en) | 2024-04-30 |
Family
ID=81197286
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111629112.XA Active CN114381363B (en) | 2021-12-28 | 2021-12-28 | Preparation method of PCR rapid detection system and PCR rapid detection system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114381363B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040062468A1 (en) * | 2002-10-01 | 2004-04-01 | National Cheng Kung University | Integrated analytical biochip and manufacturing method thereof |
| US20080160548A1 (en) * | 2005-04-20 | 2008-07-03 | Micronas Gmbh | Microoptical Detection System and Method for Determination of Temperature-Dependent Parameters of Analytes |
| CN102939161A (en) * | 2010-04-30 | 2013-02-20 | 比格科技私人有限公司 | A non contact real time micro polymerase chain reaction system and method thereof |
| CN108614600A (en) * | 2018-07-13 | 2018-10-02 | 中国科学院天津工业生物技术研究所 | A kind of high-precision chip reaction system and method |
| CN112750924A (en) * | 2020-12-31 | 2021-05-04 | 北京中科优唯科技有限公司 | Preparation method of ultraviolet light-emitting diode chip with self-photonic crystal structure |
| CN113506844A (en) * | 2021-09-08 | 2021-10-15 | 深圳市思坦科技有限公司 | Micro LED chip preparation method, micro LED chip, display device and light-emitting device |
| CN113745376A (en) * | 2021-09-08 | 2021-12-03 | 深圳市思坦科技有限公司 | Light emitting chip processing method, light emitting chip assembly, display device and light emitting device |
-
2021
- 2021-12-28 CN CN202111629112.XA patent/CN114381363B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040062468A1 (en) * | 2002-10-01 | 2004-04-01 | National Cheng Kung University | Integrated analytical biochip and manufacturing method thereof |
| US20080160548A1 (en) * | 2005-04-20 | 2008-07-03 | Micronas Gmbh | Microoptical Detection System and Method for Determination of Temperature-Dependent Parameters of Analytes |
| CN102939161A (en) * | 2010-04-30 | 2013-02-20 | 比格科技私人有限公司 | A non contact real time micro polymerase chain reaction system and method thereof |
| CN108614600A (en) * | 2018-07-13 | 2018-10-02 | 中国科学院天津工业生物技术研究所 | A kind of high-precision chip reaction system and method |
| CN112750924A (en) * | 2020-12-31 | 2021-05-04 | 北京中科优唯科技有限公司 | Preparation method of ultraviolet light-emitting diode chip with self-photonic crystal structure |
| CN113506844A (en) * | 2021-09-08 | 2021-10-15 | 深圳市思坦科技有限公司 | Micro LED chip preparation method, micro LED chip, display device and light-emitting device |
| CN113745376A (en) * | 2021-09-08 | 2021-12-03 | 深圳市思坦科技有限公司 | Light emitting chip processing method, light emitting chip assembly, display device and light emitting device |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114381363B (en) | 2024-04-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20200335665A1 (en) | Textured optoelectronic devices and associated methods of manufacture | |
| CN100587978C (en) | Solar cell and manufacturing method thereof | |
| US7210819B2 (en) | Light emitting diodes for high AC voltage operation and general lighting | |
| JP5002408B2 (en) | Lighting device | |
| Yu et al. | High-integration and high-performance micro thermoelectric generator by femtosecond laser direct writing for self-powered IoT devices | |
| CN108428782A (en) | Polysilicon nanowire thermocouple Mini-thermoelectric generator in radio frequency transceiver | |
| TW200739942A (en) | White semiconductor light emitting element and manufacturing method thereof | |
| TW200840088A (en) | Light emitting diode apparatus | |
| CN109416336B (en) | Micro heating plate device with ring structure | |
| US7550289B2 (en) | Method of fabricating an entegral device of a biochip intergrated with micro thermo-electric elements and the apparatus thereof | |
| CN205666251U (en) | Spontaneous white light emitting diode | |
| CN114381363A (en) | Preparation method of PCR (polymerase chain reaction) rapid detection system and PCR rapid detection system | |
| Xu et al. | Reduction of structural thermal resistance for deep ultraviolet light-emitting diodes fabricated on AlN ceramic substrate via copper-filled thermal holes | |
| CN105702774B (en) | A kind of self-driven schottky junction near infrared photodetector based on silicon nanowire array and preparation method thereof | |
| TW484217B (en) | White light package structure containing two LEDs | |
| CN100578828C (en) | Electroluminescent device and method for production thereof | |
| US20130026491A1 (en) | Led structure and method for manufacturing thereof | |
| CN110444559A (en) | A kind of Micro-LED array and preparation method thereof | |
| CN111863939B (en) | Bidirectional light-triggered solid-state relay integrated with silicon controlled thyristor | |
| CN213172359U (en) | Virus detection system based on micro-fluidic chip | |
| CN208898498U (en) | A kind of black silicon face infrared light supply chip of si-glass sealing | |
| CN202868631U (en) | Radiator of light-emitting diode (LED) lighting set | |
| CN217181753U (en) | Integrated LED structure | |
| CN114137324B (en) | Method for detecting quantum dot electric field effect | |
| Xu et al. | Investigation on thermal characteristics and fabrication of DUV-LEDs using copper filled thermal hole |
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 |