CN113881555A - Inferior throwing type nucleic acid isothermal amplification detection device - Google Patents
Inferior throwing type nucleic acid isothermal amplification detection device Download PDFInfo
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Abstract
The invention provides a secondary throwing type nucleic acid isothermal amplification detection device which comprises a detection part, wherein the detection part comprises a box body, a detection reaction container, a contrast reaction container and a power supply are arranged in the box body, the power supply can heat the detection reaction container and the contrast reaction container at constant temperature, the detection reaction container is provided with a detection sample inlet positioned on the outer side of the box body, the contrast reaction container is provided with a contrast sample inlet positioned on the outer side of the box body, and freeze-drying type nucleic acid amplification reagents are pre-embedded in the detection reaction container and the contrast reaction container respectively. According to the invention, the whole consumable and reagent are arranged in a disposable mode, so that the detection process can be personalized, the detection process is simple and convenient, the guidance of professionals is not needed, the detection result can be obtained in a short time, the result can be directly observed by naked eyes, and on the premise of ensuring the accuracy of the detection result, the operation is convenient, the detection cost can be reduced, and the device can more easily realize household nucleic acid detection.
Description
Technical Field
The invention belongs to the technical field of instant detection, and particularly relates to a secondary throwing type nucleic acid isothermal amplification detection device.
Background
With the improvement of living standards and the development of biotechnology, the pursuit and the attention of people to health are more and more prominent, and with the acceleration of life rhythm of people, time and efficiency become the most concerned aspects of people no matter in work or life, and particularly in the aspect of medical detection, the tedious inspection, the complex operation and the large amount of result waiting time consumption become a big problem. A large number of POCT (point-of-care testing) instruments and products are produced, such as colloidal gold detection reagents, dry chemical technology test strip biochip palm PCR instruments and the like, and the POCT instrument has the advantages of convenience in operation and quickness in detection. Based on these advantages of POCT, product instruments developed by isothermal amplification technology are widely used in agriculture, medical detection, and other industries. However, since the isothermal amplification technology is prone to aerosol contamination, misjudgment can be caused on the detection result, and higher consumable cost is often caused for single detection cost.
Disclosure of Invention
Therefore, the present invention is to provide a disposable isothermal amplification nucleic acid detecting device, which is disposable, low in material cost, accurate and fast in detection, and easy to interpret the result.
In order to solve the problems, the invention provides a secondary throwing type nucleic acid isothermal amplification detection device which comprises a detection part, wherein the detection part comprises a box body, a detection reaction container, a contrast reaction container and a power supply are arranged in the box body, the power supply can heat the detection reaction container and the contrast reaction container at constant temperature, the detection reaction container is provided with a detection sample inlet positioned on the outer side of the box body, the contrast reaction container is provided with a contrast sample inlet positioned on the outer side of the box body, and freeze-drying type nucleic acid amplification reagents are respectively pre-embedded in the detection reaction container and the contrast reaction container.
In some embodiments, the cartridge is configured with a detection observation window at a position corresponding to the detection reaction vessel, and the cartridge is configured with a control observation window at a position corresponding to the control reaction vessel.
In some embodiments, the detection reaction vessel and the control reaction vessel are wound with conductive copper wires on outer peripheral sides thereof, and the conductive copper wires are connected to both poles of the power supply.
In some embodiments, the detection reaction container and the control reaction container are further provided with heat conduction materials and/or heat conduction pastes on the outer peripheral sides.
In some embodiments, the detection reaction container comprises a first detection capillary and a second detection capillary, wherein the first detection capillary has a larger diameter than the second detection capillary, and the second detection capillary is inserted into the first detection capillary through the opening of the first detection capillary, and the detection sample can enter the first detection capillary through the opening of the second detection capillary.
In some embodiments, the control reaction container comprises a first control capillary and a second control capillary, wherein the first control capillary has a larger bore than the second control capillary, and the second control capillary is inserted into the first control capillary through the mouth of the first control capillary, and a control sample can enter the first control capillary through the mouth of the second control capillary.
In some embodiments, the lyophilized nucleic acid amplification reagents are pre-embedded within the first detection capillary and the first control capillary.
In some embodiments, a power switch and/or a heating indicator light are further disposed on the box body.
In some embodiments, the outer side of the box body is also provided with a color sample card.
In some embodiments, a sample injection dropper is further included.
The disposable nucleic acid isothermal amplification detection device provided by the invention overcomes the difficulty that the nucleic acid detection in the prior art is difficult to realize without equipment and field limitation, the whole consumable and reagent are disposable, so that the detection process can be personalized, the detection process is simple and convenient, the detection result can be obtained in a short time without the guidance of professional personnel, the result can be directly observed by naked eyes, the operation is convenient on the premise of ensuring the accuracy of the detection result, the detection cost can be reduced, and the device can more easily realize household nucleic acid detection.
Drawings
FIG. 1 is a schematic diagram showing the internal structure of a detecting part in the isothermal amplification detecting apparatus for disposable nucleic acids according to the embodiment of the present invention;
FIG. 2 is a schematic diagram showing the external structure of a detecting part in the isothermal amplification detecting apparatus for secondary polished nucleic acid according to the embodiment of the present invention;
FIG. 3 is a schematic diagram of an external structure of a sample injection dropper in the sub-disposable isothermal nucleic acid amplification detection apparatus according to the embodiment of the present invention.
The reference numerals are represented as:
1. a detection section; 11. a box body; 12. a power switch; 13. heating an indicator light; 14. a color sample card; 21. detecting the reaction vessel; 211. detecting a sample inlet; 212. detecting an observation window; 22. a control reaction vessel; 221. a reference sample inlet; 222. comparing the observation windows; 3. a power source; 4. a conductive copper wire; 5. a heat transfer device; 61. a first detection capillary; 62. a second detection capillary; 63. a first control capillary; 64. a second control capillary; 7. a lyophilized nucleic acid amplification reagent; 8. a sample injection dropper.
Detailed Description
Referring to fig. 1 to 3 in combination, according to an embodiment of the present invention, a disposable nucleic acid isothermal amplification detection apparatus is provided, including a detection component 1, the detection component 1 includes a box body 11, a detection reaction container 21, a control reaction container 22 and a power supply 3 are disposed in the box body 11, the power supply 3 can heat the detection reaction container 21 and the control reaction container 22 at a constant temperature, the detection reaction container 21 has a detection sample inlet 211 located outside the box body 11, the control reaction container 22 has a control sample inlet 221 located outside the box body 11, and a freeze-drying nucleic acid amplification reagent 7 is pre-embedded in each of the detection reaction container 21 and the control reaction container 22. In the technical scheme, the detection reaction container 21, the comparison reaction container 22 and the power supply 3 capable of heating the detection reaction container and the comparison reaction container at constant temperature are integrally arranged in the box body 11 of the detection component 1, so that the difficult problems that in the prior art, nucleic acid detection is difficult to realize without equipment and without field limitation are overcome, the whole consumable and reagent are arranged in a disposable mode, the detection process can be personalized, the detection process is simple and convenient, the professional is not required to guide, the detection result can be obtained within a short time (30 min in a specific embodiment), the result can be directly observed by naked eyes, on the premise of ensuring the accuracy of the detection result, the operation is convenient, the detection cost can be reduced, and the device can more easily realize household nucleic acid detection. It will be appreciated that the power supply 3 may be provided with a battery of sufficient capacity and that the particular design may be tailored to enable it to be disposable in a single use.
In order to further facilitate the user to observe the detection result more intuitively, in some embodiments, a detection observation window 212 is configured at a position of the cartridge 11 corresponding to the detection reaction container 21, and a control observation window 222 is configured at a position of the cartridge 11 corresponding to the control reaction container 22.
In some embodiments, the detection reaction vessel 21 and the control reaction vessel 22 are wound with conductive copper wires 4 on the outer peripheries thereof, the conductive copper wires 4 are connected to two poles of the power supply 3, and the power supply 3 generates resistance heat to the wound portions of the conductive copper wires 4, thereby achieving uniform constant temperature heating of the detection reaction vessel 21 and the control reaction vessel 22. Furthermore, a heat transfer device 5 is further provided on the outer peripheral sides of the detection reaction vessel 21 and the comparison reaction vessel 22, so that the heat generated by the heat conductive copper wires 4 can be more uniformly and constantly transferred into the respective reaction vessels, and the heat transfer device 5 may be, for example, a heat transfer paste.
As a specific implementation manner of the detection reaction container 21, the detection reaction container 21 includes a first detection capillary 61 and a second detection capillary 62, wherein the diameter of the first detection capillary 61 is larger than the diameter of the second detection capillary 62, the second detection capillary 62 is embedded in the first detection capillary 61 through the opening of the first detection capillary 61, a detection sample can enter the first detection capillary 61 through the opening of the second detection capillary 62, the second detection capillary 62 has an effect of closing a small hole, and the adverse effect of color change caused by evaporation and concentration of a reaction solution in the reaction tube and influence on result interpretation is reduced; similarly, the control reaction container 22 includes a first control capillary 63 and a second control capillary 64, wherein the diameter of the first control capillary 63 is larger than the diameter of the second control capillary 64, the second control capillary 64 is inserted into the first control capillary 63 through the opening of the first control capillary 63, and a control sample can enter the first control capillary 63 through the opening of the second control capillary 64. It is understood that, in this case, the conductive copper wire 4 is wound around the outer peripheries of the first detection capillary 61 and the first counter capillary 63, and the heat transfer device 5 is disposed on the outer peripheries of the first detection capillary 61 and the first counter capillary 63. The freeze-dried nucleic acid amplification reagent 7 is pre-embedded in the first detection capillary 61 and the first control capillary 63, and the detection sample inlet 211 and the control sample inlet 221 respectively correspond to inlets of the second detection capillary 61 and the second control capillary 64.
In some embodiments, the box 11 is further provided with a power switch 12 for controlling the constant temperature heating process of the detection component, and preferably, the box 11 is further provided with a heating indicator 13 capable of being turned on when the power switch 12 is turned on. The outer side of the box body 11 is also provided with a color sample card 14, which is marked with corresponding colors with negative or positive detection results, and the specific colors correspond to different types of reagents.
In some embodiments, a sample dropper 8 is further included for sucking the nucleic acid extract obtained by simple lysis from the detection sample inlet 211 or the control sample inlet 221. In a specific embodiment, the sample injection dropper 8 is a plastic dropper, the maximum diameter of the head of the plastic dropper is 1.5cm, and the length of the head of the plastic dropper is 2 cm; the length of the pipe part is 5cm, and the diameter of the pipe orifice is 1.5 mm. In a specific embodiment, the length, width and height of the box body 11 are 6cm × 3cm × 1.5 cm.
The operation of the apparatus for isothermal amplification of nucleic acids of the invention will be described with reference to an embodiment.
The disposable nucleic acid isothermal amplification detection device provided by the invention is used for detecting pseudomonas aeruginosa infected by clinical samples.
The method comprises the following steps:
(I) collecting a sample: the pseudomonas aeruginosa sample in this example was subjected to nucleic acid extraction of 1 alveolar lavage fluid pseudomonas aeruginosa positive sample according to the instructions in the Qingdao brevity code rapid lysis kit (section E).
(II) design of primers
The sequence of gabca gene of pseudomonas aeruginosa is adopted for analysis, isothermal amplification primers are designed aiming at specific regions, and the primer information is shown in table 1.
TABLE 1 primer sequence information for detection of Pseudomonas aeruginosa
(III) reaction system and detection step
The sample dropper 8 sucks the nucleic acid extract obtained by simple pump lysis, and slowly drops the nucleic acid extract from the detection sample inlet 211 to fill the detection observation window 212 with the sample nucleic acid extract, and the above operations are repeated to add a control sample (sterile water) from the control sample inlet 221 into the control observation window 222, wherein dry powder reagents (i.e., the freeze-dried nucleic acid amplification reagent 7, and the reagent components are shown in table 2) required for the nucleic acid amplification reaction in the embodiment are pre-embedded in the capillaries (i.e., the detection reaction container 21 and the control reaction container 22) in the detection observation window 212 and the control observation window 222. And then, turning on the power switch 12, immediately lighting the heating indicator lamp 13 to prompt the detection to start, and turning off the power switch 12 to stop heating after reacting for 30 min. After the heating is stopped for 1min, the test result is observed through the test observation window 212, and the reference color specification (i.e., the color sample card 14) is read.
TABLE 2 Dry powder reagent ingredients and concentrations used
(IV) interpretation of the test results
After the heating in the second-time throwing type nucleic acid isothermal amplification detection device is finished and the device waits for 1min, the interpretation principle of the result is as follows: 1) interpretation of the results can only be performed when the control viewing window 222 appears rosy: if the color of the reaction solution in the detection observation window 212 is yellow, the detection is positive, and if the color is rose red, the detection is negative; 2) if the control viewing window 222 appears yellow, the test is invalid and a retest is required.
In this embodiment, after the sample is heated in the second-polishing isothermal amplification detection device for 1min, the reaction solution in the control observation window 222 is rosy, and if the reaction solution in the detection observation window 212 is yellow, the sample is positive according to the interpretation principle, and thus the sample is consistent with the actual situation.
It is readily understood by a person skilled in the art that the advantageous ways described above can be freely combined, superimposed without conflict.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention. The above is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several improvements and modifications can be made without departing from the technical principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.
Claims (10)
1. The utility model provides a throw type nucleic acid isothermal amplification detection device once, a serial communication port, including determine part (1), determine part (1) includes box body (11), be provided with in box body (11) and detect reaction vessel (21), contrast reaction vessel (22) and power (3), power (3) can be right detect reaction vessel (21), contrast reaction vessel (22) constant temperature heating, detect reaction vessel (21) have and be in detection introduction port (211) in box body (11) outside, contrast reaction vessel (22) have and are in contrast introduction port (221) in box body (11) outside, pre-buried freeze-drying type nucleic acid amplification reagent (7) respectively in detect reaction vessel (21), contrast reaction vessel (22).
2. The apparatus for isothermal amplification of nucleic acid according to claim 1, wherein a detection observation window (212) is formed at a position of the cartridge (11) corresponding to the detection reaction vessel (21), and a control observation window (222) is formed at a position of the cartridge (11) corresponding to the control reaction vessel (22).
3. The isothermal amplification detection device for disposable nucleic acids according to claim 2, wherein conductive copper wires (4) are wound around the outer peripheries of the detection reaction vessel (21) and the control reaction vessel (22), and the conductive copper wires (4) are connected to both poles of the power supply (3).
4. The isothermal amplification detection apparatus for nucleic acids of the throw-less type according to claim 3, wherein a heat transfer unit (5) is further provided on the outer peripheral side of the detection reaction vessel (21) and the control reaction vessel (22).
5. The isothermal amplification detection device for disposable nucleic acids according to claim 1, wherein the detection reaction vessel (21) comprises a first detection capillary (61) and a second detection capillary (62), wherein the first detection capillary (61) has a larger diameter than the second detection capillary (62), the second detection capillary (62) is fitted into the first detection capillary (61) through the mouth of the first detection capillary (61), and a detection sample can enter the first detection capillary (61) through the mouth of the second detection capillary (62).
6. The apparatus for isothermal amplification of sub-disposable nucleic acids according to claim 1, wherein the control reaction container (22) comprises a first control capillary (63) and a second control capillary (64), wherein the first control capillary (63) has a larger diameter than the second control capillary (64), and the second control capillary (64) is inserted into the first control capillary (63) through the mouth of the first control capillary (63), and a control sample can enter the first control capillary (63) through the mouth of the second control capillary (64).
7. The isothermal amplification detection device for disposable nucleic acids according to claim 5 or 6, wherein the lyophilized nucleic acid amplification reagent (7) is pre-embedded in the first detection capillary (61) and the first control capillary (63).
8. The device for detecting the isothermal amplification of the disposable nucleic acid according to claim 1, wherein a power switch (12) and/or a heating indicator light (13) is further disposed on the box body (11).
9. The device for isothermal amplification of nucleic acid according to claim 1, wherein a color sample card (14) is further disposed on the outer side of the cassette (11).
10. The apparatus for detecting isothermal amplification of sub-disposable nucleic acids according to claim 1, further comprising a sample injection dropper (8).
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| CN202111304249.8A CN113881555A (en) | 2021-11-05 | 2021-11-05 | Inferior throwing type nucleic acid isothermal amplification detection device |
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| CN202111304249.8A CN113881555A (en) | 2021-11-05 | 2021-11-05 | Inferior throwing type nucleic acid isothermal amplification detection device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2024040739A1 (en) * | 2022-08-22 | 2024-02-29 | 凤凰医学诊断科技有限公司 | Test device |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2024040739A1 (en) * | 2022-08-22 | 2024-02-29 | 凤凰医学诊断科技有限公司 | Test device |
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