CN109082422B - Full-automatic PCR nucleic acid extraction detection device - Google Patents
Full-automatic PCR nucleic acid extraction detection device Download PDFInfo
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Abstract
The invention provides a full-automatic PCR nucleic acid extraction and detection device, which comprises: shell subassembly, microscope carrier subassembly, application of sample subassembly, supporting kit place the district, PCR fluorescence detection subassembly, control computer unit. The invention realizes the rapid and full-automatic operation of sample treatment, sample extraction and sample detection, organically integrates the separated multi-part test systems in the traditional market, can automatically and rapidly separate the samples and perform PCR detection through one-step operation, and is suitable for various samples such as blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to a full-automatic PCR nucleic acid extraction and detection device.
Background
The in vitro sample medical detection has the advantages of simplicity, automation, approaching to the site of a patient and the like, and plays an important role in the fields of clinical medicine and related medical research. At present, the automation degree of sample processing and detection is low, and the requirements of quick, timely and high-throughput sample detection in clinic cannot be met.
The nucleic acid molecule detection technology has strong advantages in inspection medicine and clinical research, and the problems of complex operation, trouble, high required cost and the like generally exist in the conventional column method, magnetic bead method and the like for nucleic acid extraction. At present, the most widely used nucleic acid detection is the real-time fluorescence PCR technology, but most of the reagents are not ready-to-use, and complex configuration work is carried out before use, so that errors and even failures of the test are easily caused. Experimental errors also arise from different personnel operations; some reagents are not stable to storage at ambient temperatures. The nucleic acid extraction detection device cannot rapidly extract nucleic acid by a one-step method; PCR detection reagents need on-site manual configuration, a large number of test samples cannot be processed in a full-automatic mode, manual marking is needed in multiple steps, detection instruments at all stages are independent, and a system device integrating the whole process of sample detection is absent.
Disclosure of Invention
In view of the above drawbacks of the prior art, the present invention provides a fully automatic PCR nucleic acid extraction and detection device. The invention provides an automatic and full-flow PCR nucleic acid extraction and detection device, which realizes the quick and full-automatic operation of sample treatment, extraction and detection, organically integrates a separated multi-part test system in the traditional market, can automatically and quickly separate a sample and perform PCR detection through one-step operation, and is suitable for various samples including blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like.
The invention provides a PCR nucleic acid extraction and detection device, which is characterized by comprising: the kit comprises a shell component, a carrier component, a sample adding component, a matched kit placing area, a PCR fluorescence detection component and a control computer device; the shell component is used for accommodating the working main body of the detection device and protecting the working main body; the sample adding component is used for realizing automatic absorption, carrying and sample adding of a sample, a PCR reaction reagent and an extracted product; the carrier assembly is used for setting the matched kit placing area and is provided with a temperature control module, a blending module and a carrying module; the temperature control module maintains proper reaction temperature; the blending module is used for automatically blending after sample adding to promote full reaction; the carrying module is used for carrying the PCR tube after the amplification reaction to the PCR fluorescence detection assembly for fluorescence detection; the first area of the matched kit placing area is a sample extracting area and is used for placing a sample container and a lysate kit; the second area of the matched kit placing area is used for placing a PCR tube; the PCR fluorescence detection component carries out fluorescence detection according to a fluorescence quantitative PCR program set by the control computer device; and the control computer device performs the control and monitoring work of absorption, carrying and sample adding and performs fluorescent quantitative PCR program setting.
Preferably, the stage assembly specifically includes: a bearing frame and a concave cavity; the concave cavity arranged on the bearing frame is used for accommodating the matched kit placing area; the concave cavity is divided into a first cavity and a second cavity, wherein the size and the shape of the first cavity are matched with those of the sample container and the lysate kit which are placed in the first area, and the size and the shape of the second cavity are matched with those of the PCR tube which is placed in the second area.
Preferably, the temperature control module of the stage assembly includes: the device comprises a first heating ring, a second heating ring, a third heating ring, a fourth heating ring, a first temperature sensor and a second temperature sensor; the first heating ring and the second heating ring surround the first cavity of the concave cavity and are used for heating the lysate kit placed in the first area so as to maintain the temperature suitable for nucleic acid lysis; the first heating ring and the second heating ring are arranged up and down, so that the first cavity is heated up and down uniformly; the first heating ring and the second heating ring are both annular electric heating pieces. The third heating ring and the fourth heating ring surround the second cavity of the concave cavity and are used for heating the PCR tube placed in the second area so as to maintain the temperature suitable for the amplification reaction; the third heating ring and the fourth heating ring are arranged up and down, so that the temperatures of the upper part and the lower part of the second cavity are uniform; the third heating ring and the fourth heating ring are also electric heating pieces; and the third heating ring and the fourth heating ring are respectively provided with an opening, and the openings of the third heating ring and the fourth heating ring are aligned with each other, so that the PCR tube is allowed to pass through the third heating ring and the fourth heating ring through the openings, and then enters and exits the second cavity.
Preferably, the carrier module comprises: a transmission driving component and a transmission sliding seat; the PCR tube is placed on the transmission sliding seat in the second area, and the transmission driving assembly comprises a torsion arm and a driving motor, wherein the transmission sliding seat is fixed with the first end of the torsion arm, and the second end of the torsion arm is connected with a motor shaft of the driving motor; therefore, the transmission sliding seat is driven to slide by the rotation of the motor shaft of the driving motor through the transmission of the torsion arm; the PCR tube slides along the conveying sliding seat, and can enter the second cavity through the openings of the third heating ring and the fourth heating ring, and the sample of the nucleic acid cracking product and the RNA enzyme removing water is loaded at the second cavity; the PCR tube can also be carried by the conveying sliding seat to move from the second cavity to the PCR fluorescent detection assembly, and the sample after the amplification reaction is subjected to fluorescent detection.
Preferably, the blending module comprises a first vibration base, a first torsion assembly, a second vibration base and a second torsion assembly; the first vibration base and the first torsion component are used for applying vibration and torsion to the lysate kit in the first cavity, so that a sample added into the kit is promoted to be fully and uniformly mixed with lysate; the second vibration base and the second torsion component drive the PCR tube in the second cavity to realize vibration and torsion, so that the PCR reagent in the frozen dry powder form, the cracked nucleic acid product and the RNA enzyme removing water are uniformly mixed.
Preferably, the sample adding assembly comprises a mechanical arm with X-Y-Z three-axial motion and an automatic sample adding gun; the mechanical arm is used for carrying the automatic sample adding gun to contact with a suction target; the automatic sample adding gun is used for realizing suction in a contact state and dripping in a non-contact state.
Preferably, the mechanical arm is divided into an upper arm, a lower arm and a support rod, wherein the upper arm is connected with the support rod through an X-Y-Z three-axis omnidirectional joint, and the upper arm can adjust the angle in the X-Y-Z three-axis direction, so that the automatic sample adding gun is aligned to a target to be sucked and dripped; the lower arm is connected with the upper arm through a one-way sliding joint, so that the lower arm can slide up and down relative to the upper arm to adjust the height of the head end of the automatic sample adding gun.
Preferably, the automatic sample adding gun comprises a gun seat and a sliding gun head, the sliding gun head is positioned below the gun seat, and a buffer part is arranged between the gun seat and the sliding gun head, so that effective buffer effect is realized.
Preferably, a fixing head is arranged on one side of the sliding gun head, which is back to the gun seat, and a pipette is arranged on the fixing head; the fixed head is communicated with a vacuum liquid driving system; the vacuum liquid-driving system comprises a liquid storage cavity and a vacuum suction injector.
Preferably, the PCR fluorescence detection assembly comprises a luminescence detection cavity, the exterior of the luminescence detection cavity is omnidirectionally surrounded by a micropore closing plate made of black light absorption material, the micropore closing plate is provided with light-transmitting micropores, and light signals generated in the luminescence detection cavity are output through the light-transmitting micropores; the light measuring instrument is used for sensing the intensity of the light signal output by the light-transmitting micropores; the light-transmitting micro-hole and the optical signal input port of the optical measuring instrument are connected to each other through an optical fiber conductive member.
Therefore, the invention realizes the rapid full-automatic operation of sample treatment, sample extraction and detection, can complete the real-time fluorescent nucleic acid molecule detection of various samples in one step by matching with a lysate kit and a PCR tube, and is suitable for various samples including blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like. The bearing frame can provide the temperature suitable for nucleic acid cracking and amplification reaction, and automatically realize the operation of mixing and conveying; the sample adding assembly can flexibly realize sample absorption and addition, and has a buffer structure, so that damage caused by hard contact is avoided, and the pipette is convenient to update; the fluorescent detection process can support digital quantitative identification, and has the advantages of accurate identification result and high identification speed.
Drawings
FIG. 1 is a schematic diagram of the PCR nucleic acid extraction and detection device according to the preferred embodiment of the present invention.
FIG. 2 is a schematic view of a carrier part of a PCR nucleic acid extraction detecting apparatus according to a preferred embodiment of the present invention.
FIG. 3 is a schematic diagram of the carrier module and the mixing module of the PCR nucleic acid extraction and detection device according to the preferred embodiment of the present invention.
FIG. 4 is a schematic view of an automatic sample application gun of the PCR nucleic acid extraction and detection device according to the preferred embodiment of the present invention;
FIG. 5 is a schematic diagram of the fluorescence detection assembly of the PCR nucleic acid extraction and detection apparatus according to the preferred embodiment of the present invention.
Detailed Description
The technical scheme of the invention is further specifically described by the following embodiments.
FIG. 1 is a schematic diagram of the structure of a fully automatic PCR nucleic acid extraction and detection device according to a preferred embodiment of the present invention. The full-automatic PCR nucleic acid extraction and detection device comprises: the kit comprises a shell component 1, a carrier component 2, a sample adding component 3, a matched kit placing area 4, a PCR fluorescence detection component 5 and a control computer device. The housing assembly 1 is used for accommodating a working body of the detection device and protecting the working body. The sample adding component 3 is used for realizing automatic absorption, carrying and sample adding of samples, PCR reaction reagents and extraction products. The carrier assembly 2 is used for arranging the matched kit placing area 4 and is provided with a temperature control module, a blending module and a carrying module; the temperature control module maintains proper reaction temperature; the blending module is used for automatically blending after sample adding to promote full reaction; the carrying module is used for carrying the PCR tube after the amplification reaction to the PCR fluorescent detection assembly 5 for fluorescent detection. The kit placement region 4 includes the following regions: the first area is a sample extraction area and is used for placing a sample container and a lysate kit, wherein nucleic acid lysate is arranged in the lysate kit, and a sample to be detected, such as blood and the like, is sampled from the sample container and then added to the lysate kit by a sample adding assembly 3, so that the nucleic acid can be cracked, and the nucleic acid sample is completely released; the second area is a PCR tube storing a freeze-dried PCR reagent, the freeze-dried powder PCR reagent can be directly used for fluorescent quantitative PCR measurement after sample loading, and the PCR reagent comprises various types of PCR reaction buffer solutions, DNA polymerase, various reaction specific primers and probes to form a PCR amplification system; the sample adding component 3 directly adds the cracked nucleic acid product and the RNA enzyme removing water in the first area into the PCR tube in the second area, the mixture is automatically and uniformly mixed through the uniformly mixing module of the carrier component 2, and finally the obtained liquid in the PCR tube can be directly used for fluorescence detection. And the PCR fluorescence detection component 5 carries out fluorescence detection according to a fluorescence quantitative PCR program set by the control computer device. And the control computer device performs the control and monitoring work of absorption, carrying and sample adding and performs fluorescent quantitative PCR program setting.
The nucleic acid rapid extraction detection kit matched with the device comprises two parts, namely a lysate kit and a PCR tube, wherein the lysate kit is placed in the first area; a PCR tube is placed in the second region. The lysis solution reagent box is filled with nucleic acid lysis solution, a sample to be detected is added into the lysis solution reagent box through the sample adding component 3, the virus structure can be changed under the action of the lysis solution, virus nucleic acid is released, the lysis solution does not contain components for inhibiting real-time fluorescence PCR reaction, and the product can be directly used for subsequent PCR detection. The frozen dry powder PCR amplification system reagent directly used for fluorescent quantitative PCR measurement is placed in the PCR tube, and is an obtained product prepared by using various PCR reaction buffer solutions, DNA polymerase, various reaction specific primers, probes and freeze-drying protective agents as raw materials through a freeze-drying process. The sample adding component 3 adds the cracked nucleic acid product in the first area into the PCR tube, RNA enzyme removing water is added into the PCR tube, and fluorescence detection can be directly executed by the PCR fluorescence detection component 5 through automatic mixing of the even mixing module of the carrier component 2.
As shown in fig. 2 to 3, the stage assembly 2 specifically includes: the device comprises a bearing frame 201, a concave cavity 202, a first heating ring 203, a second heating ring 204, a third heating ring 205, a fourth heating ring 206, a first temperature sensor 207, a second temperature sensor 208, a transmission driving assembly 209, a transmission sliding seat 210, a first vibration base 211, a first torsion assembly 212, a second vibration base 213 and a second torsion assembly 214.
As shown in fig. 2, the concave cavity 202 of the carrier 201 is used for accommodating the kit holding area 4, the kit holding area 4 is divided into a first area and a second area, and thus the concave cavity 202 is also divided into a first cavity and a second cavity, wherein the size and shape of the first cavity are matched with the size and shape of the sample container and the lysate kit placed in the first area, and the size and shape of the second cavity are matched with the size and shape of the PCR tube placed in the second area. The temperature control module includes: a first heating ring 203, a second heating ring 204, a third heating ring 205, a fourth heating ring 206, a first temperature sensor 207, and a second temperature sensor 208. The first heating ring 203 and the second heating ring 204 surround the first cavity of the concave cavity 202, and are used for heating the lysate kit placed in the first area so as to maintain the temperature suitable for nucleic acid lysis; the first heating ring 203 and the second heating ring 204 are arranged up and down, so that the first cavity is heated up and down uniformly; the first heating ring 203 and the second heating ring 204 are both annular electric heating sheets. A third heating ring 205 and a fourth heating ring 206 surround the second cavity of the concave cavity 202, and are used for heating the PCR tube placed in the second region, so as to maintain a temperature suitable for the amplification reaction; the third heating ring 205 and the fourth heating ring 206 are also arranged up and down, so that the upper part and the lower part of the second cavity have uniform temperature; moreover, the third heating ring 205 and the fourth heating ring 206 are also both electric heating sheets; and the third heating ring 205 and the fourth heating ring 206 are not closed rings, but have an opening respectively, and the openings of the third heating ring 205 and the fourth heating ring 206 are aligned with each other, so that the PCR tube is allowed to pass through the third heating ring 205 and the fourth heating ring 206 via the openings, and then enters and exits from the second cavity, so that the PCR tube is carried between the stage assembly 2 and the PCR fluorescence detection assembly 5. The first temperature sensor 207 is used for monitoring the real-time temperature in the first cavity, and adjusting the heating values of the first heating ring 203 and the second heating ring 204 according to the monitored temperature value, so as to maintain the temperature suitable for nucleic acid lysis. The second temperature sensor 208 is used for monitoring the real-time temperature of the second chamber, and adjusting the heating values of the third heating ring 205 and the fourth heating ring 206 according to the monitored temperature value, so as to maintain the temperature suitable for the amplification reaction.
As shown in fig. 3, the carrier module includes a transfer drive assembly 209 and a transfer shoe 210. The PCR tube is placed on the transfer slide block 210 in the second area, and the transfer drive assembly 209 includes a torsion arm 209A and a drive motor 209B, wherein the transfer slide block 210 is fixed to a first end of the torsion arm 209A, and a second end of the torsion arm 209A is connected to a motor shaft of the drive motor 209B. Accordingly, the transmission slide base 210 is driven to slide by the rotation of the motor shaft of the driving motor 209B through the torsion arm 209A. The PCR tube is carried by the conveying sliding seat 210 to slide, and can enter the second cavity through the openings of the third heating ring 205 and the fourth heating ring 206, and the sample of the nucleic acid cleavage product and the RNA enzyme removing water is loaded at the second cavity; the PCR tube can also be carried by the transmission slide seat 210 to move from the second cavity to the PCR fluorescence detection assembly 5, and the sample after the amplification reaction is subjected to fluorescence detection.
The blending module comprises a first vibration base 211, a first torsion component 212, a second vibration base 213 and a second torsion component 214. The first vibration base 211 and the first torsion component 212 are used for applying vibration and torsion to the lysate reagent kit in the first cavity, so that a sample added with the reagent kit is promoted to be fully and uniformly mixed with lysate; the first vibration base 211 is powered by an eccentric vibration motor to realize vibration, and the first torsion assembly 212 is driven by a torsion motor to perform torsion in a certain angle range in a reciprocating manner. The second vibrating base 213 and the second twisting component 214 drive the PCR tube in the second cavity to vibrate and twist, so as to promote the PCR reagent in the form of frozen dry powder to be uniformly mixed with the cracked nucleic acid product and the rnase-removing water.
The sample adding component 3 samples a sample to be detected, such as blood and the like, from a sample container, then adds the sample to the lysate kit, and adds a cracked nucleic acid product and RNA enzyme removing water into the PCR tube. The sample adding component 3 comprises a mechanical arm 301 with X-Y-Z three-axial motion and an automatic sample adding gun 302. As shown in fig. 1, the mechanical arm 301 is divided into an upper arm 301A, a lower arm 301B and a support rod 301C, wherein the upper arm 301A and the support rod 301C are connected by an omnidirectional joint with three axes X-Y-Z, so that the upper arm 301A can be adjusted in angle in the three axes X-Y-Z to align the automatic sample application gun 302 with the target to be aspirated and dropped (the target includes a sample container, a lysate kit and a PCR tube). The lower arm 301B is connected with the upper arm 301A through a one-way sliding joint, so that the lower arm 301B can slide up and down relative to the upper arm 301A to adjust the height of the head end of the automatic sample adding gun 302, contact between the automatic sample adding gun 302 and a suction target is realized, suction is realized in a contact state, and then the separation is realized by moving up.
As shown in fig. 4, the automatic sample adding gun 302 is provided with a gun holder 302A and a slide gun head 302B, and the slide gun head 302B is located below the gun holder 302A and a buffer portion 302C is provided therebetween. Buffer 302C is installed between gun seat 302A and slide gun head 302B, realizes effectual cushioning effect, and when carrying out the application of sample, when making automatic application of sample rifle 302 contact target through manipulator 301, automatic application of sample rifle 302 continues down, and buffer 302C plays a role this moment, can make effectively laminating between automatic application of sample rifle 302 and the target. The buffer portion 302C specifically includes a guide post 302G and a buffer spring 302H sleeved on the guide post 302G, one end of the guide post 302G is fixedly connected with the sliding gun head 302B, the other end of the guide post 302G passes through the gun base 302A to be in sliding fit with the gun base, and the buffer spring 302H is disposed between the gun base 302A and the sliding gun head 302B. A fixing head 302D is arranged on one side of the sliding gun head 302B, which faces away from the gun base 302A, a pipette 302E is mounted on the fixing head 302D, and the fixing head 302D is communicated with a vacuum liquid driving system 302F. The vacuum flood system 302F includes a fluid reservoir 302I and a vacuum suction syringe 302J; when the suction operation is performed, the vacuum suction syringe 302J provides a vacuum suction force to suck the liquid into the liquid storage cavity 302I through the fixing head 302D for storage; during the dropping operation, the vacuum suction syringe 302J provides a pushing force to push the liquid out of the liquid storage chamber 302I through the fixing head 302D. The pipette 302E is coupled to the fixed head 302D by the sliding sleeve 302K, so that automatic removal of the pipette 302E from the fixed head 302D can be achieved by moving the sliding sleeve 302K upward.
After the sample adding component 3 adds the cracked nucleic acid product and the RNA enzyme removing water into the PCR tube and fully and uniformly mixes the products to complete the amplification reaction, the PCR tube is transmitted to the PCR fluorescence detection component 5 by the carrying module, and the PCR fluorescence detection component 5 performs fluorescence detection according to the fluorescence quantitative PCR program set by the control computer device. The PCR fluorescence detection assembly 5 comprises a luminescence detection cavity 501, and the outside of the luminescence detection cavity 501 is omnidirectionally surrounded by a micropore closing plate 502 made of black light absorption materials, so that the interference effect of external light is avoided. The micro-hole closing plate 502 has a light-transmissive micro-hole 503 through which a light signal generated in the luminescence detection chamber 501 is output. The light meter 504 is used for sensing the intensity of the light signal output through the light-transmissive micro-hole 503. The light-transmitting micro-hole 503 and the optical signal input port of the optical measurement instrument 504 are connected to each other through an optical fiber conducting member 505, and the optical measurement instrument further includes a photoelectric gain converter, a signal amplifier, and an analog-to-digital conversion circuit, where the photoelectric gain converter and the signal amplifier are used to convert and amplify an optical signal into an electrical signal, and the electrical signal is further digitized by the analog-to-digital conversion circuit, and finally, a digital signal representing the intensity of the optical signal generated by signal acquisition, amplification, and sampling quantization is output. In one measurement, the intensity of the optical signal is acquired repeatedly. The control computer device is used for receiving the digital signal sent by the optical measuring instrument 504 and generating a luminous curve corresponding to the sample to be measured based on a large amount of optical signal intensity, wherein the luminous curve represents the relationship between the optical signal intensity and time. And the control computer device also prestores a plurality of standard curves, and each standard curve corresponds to a reference value. The luminescence curve and the standard curve are fitted to produce a measured value.
Therefore, the invention realizes the rapid full-automatic operation of sample treatment, sample extraction and detection, can complete the real-time fluorescent nucleic acid molecule detection of various samples in one step by matching with a lysate kit and a PCR tube, and is suitable for various samples including blood, serum, plasma, saliva, urine, interstitial fluid, semen, secretion, pus, respiratory fluid, mucus and the like. The bearing frame can provide the temperature suitable for nucleic acid cracking and amplification reaction, and automatically realize the operation of mixing and conveying; the sample adding assembly can flexibly realize sample absorption and addition, and has a buffer structure, so that damage caused by hard contact is avoided, and the pipette is convenient to update; the fluorescent detection process can support digital quantitative identification, and has the advantages of accurate identification result and high identification speed.
The above embodiments are only for illustrating the invention and are not to be construed as limiting the invention, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention, therefore, all equivalent technical solutions also belong to the scope of the invention, and the scope of the invention is defined by the claims.
Claims (2)
1. A full-automatic PCR nucleic acid extraction and detection device is characterized by comprising: the kit comprises a shell component, a carrier component, a sample adding component, a matched kit placing area, a PCR fluorescence detection component and a control computer device; the shell component is used for accommodating the working main body of the detection device and protecting the working main body; the sample adding component is used for realizing automatic absorption, carrying and sample adding of a sample, a PCR reaction reagent and an extracted product; the carrier assembly is used for setting the matched kit placing area and is provided with a temperature control module, a blending module and a carrying module; the temperature control module maintains proper reaction temperature; the blending module is used for automatically blending after sample adding to promote full reaction; the carrying module is used for carrying the PCR tube after the amplification reaction to the PCR fluorescence detection assembly for fluorescence detection; the first area of the matched kit placing area is a sample extracting area and is used for placing a sample container and a lysate kit; the second area of the matched kit placing area is used for placing a PCR tube; the PCR fluorescence detection component carries out fluorescence detection according to a fluorescence quantitative PCR program set by the control computer device; the control computer device performs the control and monitoring work of absorption, carrying and sample adding and performs the fluorescent quantitative PCR program setting;
wherein, the microscope carrier subassembly specifically includes: a bearing frame and a concave cavity; the concave cavity arranged on the bearing frame is used for accommodating the matched kit placing area; the concave cavity is divided into a first cavity and a second cavity, wherein the size and the shape of the first cavity are matched with those of the sample container and the lysate kit which are placed in the first area, and the size and the shape of the second cavity are matched with those of the PCR tube which is placed in the second area;
the temperature control module of the carrier assembly comprises: the device comprises a first heating ring, a second heating ring, a third heating ring, a fourth heating ring, a first temperature sensor and a second temperature sensor; the first heating ring and the second heating ring surround the first cavity of the concave cavity and are used for heating the lysate kit placed in the first area so as to maintain the temperature suitable for nucleic acid lysis; the first heating ring and the second heating ring are arranged up and down, so that the first cavity is heated up and down uniformly; the first heating ring and the second heating ring are both annular electric heating sheets, and the third heating ring and the fourth heating ring surround the second cavity of the concave cavity and are used for heating the PCR tube placed in the second area so as to maintain the temperature suitable for amplification reaction; the third heating ring and the fourth heating ring are arranged up and down, so that the temperatures of the upper part and the lower part of the second cavity are uniform; the third heating ring and the fourth heating ring are also electric heating pieces; and the third heating ring and the fourth heating ring are respectively provided with an opening, the openings of the third heating ring and the fourth heating ring are aligned with each other, so that the PCR tube is allowed to pass through the third heating ring and the fourth heating ring through the openings, and then enters and exits the second cavity;
the carrier module comprises: a transmission driving component and a transmission sliding seat; the PCR tube is placed on the transmission sliding seat in the second area, and the transmission driving assembly comprises a torsion arm and a driving motor, wherein the transmission sliding seat is fixed with the first end of the torsion arm, and the second end of the torsion arm is connected with a motor shaft of the driving motor; therefore, the transmission sliding seat is driven to slide by the rotation of the motor shaft of the driving motor through the transmission of the torsion arm; the PCR tube slides along the conveying sliding seat, and can enter the second cavity through the openings of the third heating ring and the fourth heating ring, and the sample of the nucleic acid cracking product and the RNA enzyme removing water is loaded at the second cavity; the PCR tube can also be carried by the conveying sliding seat to move from the second cavity to the PCR fluorescence detection assembly, and the sample after the amplification reaction is subjected to fluorescence detection;
the blending module comprises a first vibration base, a first torsion assembly, a second vibration base and a second torsion assembly; the first vibration base and the first torsion component are used for applying vibration and torsion to the lysate kit in the first cavity, so that a sample added into the kit is promoted to be fully and uniformly mixed with lysate; the second vibration base and the second torsion assembly drive the PCR tube in the second cavity to realize vibration and torsion, so that the PCR reagent in the frozen dry powder form is uniformly mixed with the cracked nucleic acid product and the RNA enzyme-removed water;
the sample adding assembly comprises a mechanical arm with X-Y-Z three-axial motion and an automatic sample adding gun; the mechanical arm is used for carrying the automatic sample adding gun to contact with a suction target; the automatic sample adding gun is used for realizing absorption in a contact state and dripping in a non-contact state; the automatic sample adding gun comprises a gun seat and a sliding gun head, the sliding gun head is positioned below the gun seat, and a buffer part is arranged between the gun seat and the sliding gun head, so that an effective buffer effect is realized; the buffering part comprises a guide pillar and a buffering spring sleeved on the guide pillar in a penetrating manner, one end of the guide pillar is fixedly connected with the sliding gun head, the other end of the guide pillar penetrates through the gun base to be in sliding fit with the gun base, and the buffering spring is arranged between the gun base and the sliding gun head; a fixing head is arranged on one side of the sliding gun head, back to the gun base, and a pipette is arranged on the fixing head; the fixed head is communicated with a vacuum liquid driving system; the vacuum liquid driving system comprises a liquid storage cavity and a vacuum suction injector, and when suction operation is executed, the vacuum suction injector provides vacuum suction force to suck liquid into the liquid storage cavity through the fixing head for storage; when the dropping operation is carried out, the vacuum suction injector provides thrust to push the liquid out of the liquid storage cavity through the fixing head; the pipette is combined with the fixed head through the sliding sleeve, so that the pipette can be automatically dismounted from the fixed head by moving the sliding sleeve upwards;
the PCR fluorescence detection assembly comprises a luminescence detection cavity, the exterior of the luminescence detection cavity is surrounded by a micropore closing plate made of black light absorption material in an omnidirectional way, the micropore closing plate is provided with light-transmitting micropores, and light signals generated in the luminescence detection cavity are output through the light-transmitting micropores; the light measuring instrument is used for sensing the intensity of the light signal output by the light-transmitting micropores; the light-transmitting micropore and the optical signal input port of the optical measuring instrument are connected with each other through an optical fiber conducting component; the optical measuring instrument also comprises a photoelectric gain converter, a signal amplifier and an analog-to-digital conversion circuit, wherein the photoelectric gain converter and the signal amplifier are used for converting an optical signal into an electric signal and amplifying the electric signal, the electric signal is further digitized by the analog-to-digital conversion circuit, and finally, a digital signal which is generated by signal acquisition, amplification and sampling quantization and represents the intensity of the optical signal is output;
in one measurement, the light measuring instrument obtains the intensity of the light signal through repeated collection, the control computer device is used for receiving the digital signal sent by the light measuring instrument, a luminous curve corresponding to a sample to be measured is generated based on a large amount of light signal intensities, the luminous curve represents the relation between the light signal intensity and time, the control computer device also prestores a plurality of standard curves, each standard curve corresponds to a reference value, and the luminous curve and the standard curves are fitted to generate a measurement value.
2. The fully automatic PCR nucleic acid extraction and detection device according to claim 1, wherein the mechanical arm is divided into an upper arm, a lower arm and a support rod, wherein the upper arm and the support rod are connected through an X-Y-Z three-axis omnidirectional joint, and the upper arm can be adjusted in angle in the X-Y-Z three-axis direction so that the automatic sample adding gun is aligned with the target to be sucked and dropped; the lower arm is connected with the upper arm through a one-way sliding joint, so that the lower arm can slide up and down relative to the upper arm to adjust the height of the head end of the automatic sample adding gun.
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| EP3995565A4 (en) * | 2019-07-01 | 2022-11-16 | Shenyi Biotech (Hangzhou) Co., Ltd. | Molecular detection system |
| CN110331090A (en) * | 2019-07-12 | 2019-10-15 | 湖南圣湘生物科技有限公司 | Nucleic acid extraction, amplification and detection device |
| CN111421560A (en) * | 2020-04-10 | 2020-07-17 | 前元运立(北京)机器人智能科技有限公司 | Isolation space virus diagnosis robot system |
| CN111534423A (en) * | 2020-05-12 | 2020-08-14 | 西安交通大学 | Group nucleic acid testing and mixing device and method |
| CN115885032A (en) * | 2020-08-28 | 2023-03-31 | 深圳华大智造科技股份有限公司 | Integrated intelligent nucleic acid detection system |
| CN111979094A (en) * | 2020-08-28 | 2020-11-24 | 中国科学院苏州生物医学工程技术研究所 | Nucleic acid detection device |
| CN113308368B (en) * | 2021-06-15 | 2024-05-03 | 中国科学院苏州生物医学工程技术研究所 | Full-automatic nucleic acid detection device |
| CN114480101B (en) * | 2022-04-01 | 2022-12-13 | 安永医疗科技常州有限公司 | Rotary nucleic acid extraction and detection device |
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