CN218755755U - Molecule detection device suitable for field detection - Google Patents

Abstract

The invention relates to the field of molecular biological detection, in particular to a molecular detection device and a method suitable for field detection, wherein the molecular detection device comprises: a sample processing container, an extraction test cassette and a heating platform, the extraction test cassette comprising a nucleic acid extraction zone and a detection zone; wherein the nucleic acid extraction region comprises: the sample cavity is provided with an injection hole and is communicated with the nucleic acid binding cavity in a fluid way, and the nucleic acid binding cavity is respectively communicated or closed with the waste liquid cavity and the collection cavity through a valve; the detection zone includes: the reaction cavity is communicated with the collection cavity in a fluid mode, and the reaction cavity is provided with an injection hole. The detection device of the invention can complete sampling, nucleic acid extraction, amplification and detection at one time, can carry out molecular detection on biological samples within 30min, is suitable for field detection, and is convenient and quick.

Description

Molecule detection device suitable for field detection

Technical Field

The utility model relates to a molecular biology detects the field, especially relates to a molecular detection device suitable for witnessed inspections.

Background

With the rapid development of modern biotechnology, molecular information in a sample is obtained by using a molecular detection technology, and the molecular information is widely used. In some cases, in order to obtain the detection result timely and quickly, the sample needs to be detected on site. The molecular detection comprises three necessary steps of nucleic acid extraction, nucleic acid amplification and product monitoring, and the DNA extraction methods commonly used in laboratories at present comprise a CTAB method, an SDS method, a kit method and the like. The nucleic acid extraction methods need to use experimental equipment such as a centrifuge, a pipettor and the like, and are complex to operate and long in time consumption. Nucleic acid amplification requires temperature elevation, is complicated to operate, requires expensive materials and equipment, and is difficult to directly apply to in-situ detection.

Isothermal nucleic acid amplification, such as loop-mediated isothermal amplification ("LAMP"), has been widely used because it requires only a constant temperature device (e.g., a water bath) to perform a rapid and efficient amplification reaction, and requires less instrumentation. And in order to meet the requirements of convenience and quickness of field detection, a plurality of technical and related device adaptive improvements are made. For example, CN209537484U discloses a device for rapid detection of nucleic acid amplification, which comprises a sealed bottle 1, wherein a reaction insert tube 2 is arranged in the sealed bottle 1, in an experiment for rapid detection of nucleic acid amplification, 100ul of a system is added into the reaction insert tube 2, a bottle cap 102 is screwed to keep the sealed bottle 1 sealed, and the sealed bottle is put into a constant temperature environment of 63 ℃ for a bath for 60min, so that the amplification system reacts. Although the device can simply and conveniently realize amplification, the detection process in the field at the outdoor at least involves the steps of sample treatment, reagent control and the like, and the device cannot be used for realizing the amplification.

SUMMERY OF THE UTILITY MODEL

The utility model provides a molecular detection device suitable for witnessed inspections combines nucleic acid amplification and micro-fluidic technique, can realize the function that nucleic acid drawed and gene amplification simultaneously on micro-fluidic chip, will sample, nucleic acid drawing, amplification, detect once only to accomplish, can carry out molecular detection to biological sample in 30min, is suitable for witnessed inspections.

The utility model adopts the following technical proposal:

a molecular testing device adapted for in situ testing, comprising:

a sample processing container for performing cell lysis processing on a sample;

an extraction test cassette comprising a nucleic acid extraction zone and a detection zone;

a heating platform for heating the whole or the detection area part of the extraction detection box to reach the reaction temperature of nucleic acid amplification;

wherein the nucleic acid extraction region comprises: a sample cavity, a nucleic acid combining cavity, a waste liquid cavity and a collection cavity, wherein the sample cavity is provided with an injection hole and is in fluid communication with the nucleic acid combining cavity, and the nucleic acid combining cavity is respectively in fluid communication or closed with the waste liquid cavity and the collection cavity through valves; an extraction carrier for combining nucleic acid molecules is arranged in the nucleic acid combining cavity;

the detection zone includes: the reaction cavity is communicated with the collection cavity in a fluid mode, and the reaction cavity is provided with an injection hole.

In-situ testing techniques involve sample handling, reagent manipulation, and result analysis. The micro-fluidic technology is a technology for accurately controlling fluid on a micro-nano scale, can utilize small-volume liquid to realize automatic processing, quick reaction and the like of a sample and a reagent in a chip, and is very suitable for field detection application. The utility model discloses combine nucleic acid amplification and micro-fluidic technique, utilize structural ingenious design, carry out the extraction of sample nucleic acid and gene amplification integration in same device, device compact structure, small, occupation space is little, and convenient operation, simple, swiftly realize nucleic acid extraction and gene amplification.

Preferably, the nucleic acid extraction region further comprises: and the cleaning liquid cavity and the elution liquid cavity are both provided with injection holes, and the cleaning liquid cavity and the elution liquid cavity are both communicated with the collection cavity through fluid.

In this scheme, independently set up washing liquid chamber and eluent chamber, the washing liquid separately pours into with the elution liquid, can also deposit elution liquid or washing liquid in advance in the intracavity that corresponds, closes corresponding valve, need not to carry the strorage device of elution liquid and washing liquid in addition, and is more portable, during the use, only need open corresponding valve can. A sealing cover is arranged at the opening.

Preferably, the detection zone further comprises: a dosing chamber having an inlet end in fluid communication with the collection chamber and an outlet end in fluid communication with the reaction chamber. The quantitative cavity can be used for quantifying the reaction liquid, so that the quantitative requirement of detection is met.

Preferably, the detection zone is provided in plurality and the plurality of quantification chambers are each in fluid communication with the collection chamber. One-to-many high-throughput detection can be achieved.

Preferably, the heating platform has a heating slot for insertion of all or part of the extraction cartridge.

Preferably, the heating platform can be powered by a battery, a power supply or a USB interface, and can also be used for building a solar panel, and the platform is multilayer and can heat a plurality of chips simultaneously.

Preferably, the molecular detection device includes a polishing tool for polishing a sample, and is suitable for processing a sample to be polished, for example, a sample such as a fruit, a leaf, or a rhizome.

More specifically, the grinding tool includes a grinding rod and a grinding pipe, and may also include only the grinding rod, and the grinding is performed directly in the sample processing container.

Preferably, the extraction carrier is filter paper or a silica gel membrane.

As the preferred, this molecular detection device still contains sampling tool, and the sampler has the sampling tube and takes a sample and cuts, and sampling tube one end is the handle, is used for holding, and other end incision is the inclined plane, can scrape the sample that awaits measuring during the sample and take a sample, also can pierce the sample that awaits measuring inside and gather. Sampling tubes of different specifications and sizes can be selected according to the amount of required samples. When the sample is hard or dry and is not suitable for a sampling tube, the sample can be collected by using a sampling scissors, and the sample can be cut into pieces by using the sampling scissors so as to facilitate the extraction of the nucleic acid.

Preferably, the molecular detection device further comprises an extractor for extracting the sample liquid lysed in the sample processing container into the sample chamber.

Preferably, the extractor is a syringe having a capillary tube provided at the tip thereof.

When washing liquid or eluant wasing or eluting the sample through the sample chamber, owing to draw the existence of carrier for the process of washing liquid or eluant is obstructed, and the speed of flowing through is very slow, consequently preferred, the sprue hole department in sample chamber is provided with the sealing rubber circle, and the centre bore of sealing rubber circle and the capillary bore adaptation of syringe set up like this, the capillary of syringe inserts the back, can with form the state of relative sealing between the sealing rubber circle, utilize the syringe like the injection of sample intracavity, provide certain pressure, be favorable to washing liquid or eluant through the speed of sample, improve washing and elution efficiency.

Similarly, it is also preferable that a sealing rubber ring is provided in another injection hole, for example, the injection hole of the reaction chamber and the injection hole of the detection chamber, and a center hole of the sealing rubber ring is adapted to the capillary caliber of the syringe. The liquid in the corresponding cavity needs to be provided with pushing force, and then the liquid can be pushed by using the extractor, so that the liquid can smoothly enter the next procedure.

Preferably, the molecular detection device further comprises a box for accommodating original components such as the sample processing container and the extraction detection box, so that the whole device is convenient to store, store and carry. A handle is arranged on the box body.

Through implementing above-mentioned technical scheme, the utility model discloses following beneficial effect has:

utilize the utility model discloses a detection device will sample, nucleic acid extraction, amplification, detect once only to accomplish, can carry out the molecular detection to biological sample in 30min, is suitable for the witnessed inspections, and is convenient, swift.

Drawings

FIG. 1 is a plan view of a molecular detector according to an embodiment of the present invention;

FIG. 2 is a view showing an internal structure of the molecular detection device shown in FIG. 1;

FIG. 3 is a schematic structural diagram of a molecular detection device according to another embodiment of the present invention;

FIG. 4 is a top view of a molecular testing device according to another embodiment of the present invention;

FIG. 5 is a top view of the molecular detection device of the present invention suitable for one-to-many high throughput detection;

fig. 6 is a schematic structural view of a heating platform in the molecular detection device according to an embodiment of the present invention;

fig. 7 is a schematic structural view of a sample processing container in the molecular detection device according to an embodiment of the present invention.

Fig. 8 is a schematic structural diagram of an extractor in a molecular detection device according to an embodiment of the present invention.

Detailed Description

The invention is explained in more detail below with reference to the figures and the embodiments.

It should be noted that the following embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments can be modified, or some technical features can be equivalently replaced; the modifications or substitutions do not depart from the scope of the technical solutions of the embodiments of the present invention.

Example 1

This example provides a molecular detection device suitable for on-site detection, and specifically takes early detection of Pyricularia oryzae RAA-LFD as an example to perform one-to-one qualitative detection.

The molecular detection device includes: an extraction cartridge 100, a sample processing container 200, a heating stage 300, and an extractor 400.

The extraction detection box comprises a nucleic acid extraction area 1A and a detection area 1B; a transparent cover is placed over the two areas to allow the interior to be viewed. As shown in figure 1.

Referring specifically to FIG. 2, the nucleic acid extraction region 1A comprises: a sample cavity 11, a nucleic acid combining cavity 12, a waste liquid cavity 13 and a collection cavity 14, wherein the sample cavity 11 is provided with an injection hole 15, the injection hole 15 protrudes out of the cover to facilitate the injection of a sample, the sample cavity 11 is in micro fluid communication with the nucleic acid combining cavity 12 through a capillary tube 16, the nucleic acid combining cavity 12 is respectively in micro fluid communication with the waste liquid cavity 13 and the collection cavity 14 through the capillary tube 16 and is communicated or closed through a capillary valve control; an extraction carrier for binding nucleic acid molecules is arranged in the nucleic acid binding cavity 12, and the extraction carrier can be nucleic acid binding filter paper or a silica gel membrane.

The detection zone 1B includes: a reaction chamber 17 and a detection chamber 18, the reaction chamber 17 is in fluid communication with the collection chamber 14, the reaction chamber 17 is provided with an injection hole 15, the injection hole 15 of the reaction chamber 17 is used for injecting reaction liquid, the detection chamber 18 may be provided with the injection hole 15 or not, depending on the detection means, if the detection liquid is required to be used for detection, the injection hole 15 is arranged, if the detection is carried out by means of test paper and the like, the pre-buried test paper is in the detection chamber 18, and at this time, the injection hole 15 does not need to be additionally arranged.

The heating platform 300, as shown in fig. 6, the heating platform 300 has a heating base portion, a heating wire is disposed inside the heating platform, the heating portion above the heating base portion has one or more heating grooves, and a plurality of extraction detection boxes 100 can be disposed in the plurality of heating grooves, so as to achieve a plurality of heating simultaneously. The whole or part of the extraction kit 100 is inserted into the groove, and the whole or detection region 1B of the extraction kit 100 is heated to a reaction temperature for nucleic acid amplification. Only the reaction chamber 17 in the detection region 1B needs to be heated, but since the extraction cartridge 100 is generally small in size, local heating is inconvenient, and it is difficult to control the heating portion, the extraction cartridge 100 is heated as a whole.

The sample processing container 200 is a tubular container, and as shown in fig. 7, after the crushed sample is added, a cell lysate is added. There are also some samples that do not require lysis, and in this case, no lysis solution needs to be added.

The extractor 400 extracts the sample liquid lysed in the sample processing container 200 into the sample chamber 11. The extractor 400 is shown in fig. 8, and is directly a syringe with a needle at the front end, and the needle can be replaced by a suitable capillary tube 16, and the capillary tube 16 is preferably a plastic soft and bendable capillary tube 16.

This molecular detection device still contains sampling tool, and sampling tool has the sampling tube and the sample is cut, and sampling tube one end is the handle, is used for holding, and the other end incision is the inclined plane, can scrape the sample to be measured during the sample and sample, also can pierce the sample to be measured inside and gather. Sampling tubes with different specifications and sizes can be selected according to the amount of required samples. When the sample is hard or dry and is not suitable for a sampling tube, the sample can be collected by using a sampling scissors, and the sample can be cut into pieces by using the sampling scissors so as to be convenient for extracting the nucleic acid.

The molecular detection device also comprises a grinding tool, and the grinding tool is used for grinding the sample and is suitable for sample treatment needing grinding, such as samples of fruits, leaves, rhizomes and the like. The grinding tool includes a grinding rod and a grinding tube. The grinding tool is only one embodiment in this embodiment, and is not necessary for the molecular detection device of the present invention. The sample needing no grinding does not need a grinding tool, and the sample needing grinding is not necessarily required, and can be cut into pieces by sampling scissors, so that the aim can be fulfilled.

In the device, except that the extraction detection box is disposable and not reused, other elements can be reused.

The embodiment also provides a molecular detection method suitable for field detection, the detection device is adopted, the early detection of rice blast bacteria RAA-LFD is taken as an example to carry out one-to-one qualitative detection, a sampling tube is used for scraping and sampling on rice seedling leaves, and fresh rice seedling samples can be directly detected without grinding. The sample is put into the lysis solution, 5uL lysis solution is absorbed after shaking, the sample is injected into the sample cavity 11 by an extractor, the sample is controlled to flow through the nucleic acid molecule combined filter paper sheet, DNA is absorbed in the filter paper, and the rest liquid enters the waste liquid cavity 13. The cleaning fluid is then controlled by the extractor to be injected through the injection hole 15 of the sample chamber 11, through the filter paper sheet for cleaning, and through the filter paper sheet into the waste chamber 13. The extractor is then used to inject an eluent through the injection hole 15 of the sample chamber 11 to elute the DNA from the filter paper sheet and into the collection chamber 14. The DNA in the collection chamber 14 is pushed into the reaction chamber 17, and the reaction solution may be injected into the reaction chamber 17 through the injection hole 15, or the reaction solution may be sealed in the reaction chamber 17 in advance. And then heating the chip to 39 ℃ through a heating platform, maintaining for 20min to realize amplification reaction, pushing the reaction liquid to a detection area 1B by using an extractor for reaction products after the reaction is finished, injecting a buffer solution from an injection hole 15, embedding a test strip in the detection area 1B, and observing the detection line of the test strip so as to judge whether the sample contains the rice blast genes.

Example 2

The difference from example 1 is that, on the basis of example 1, a quantitative cavity 19 is added in the detection zone 1B, as shown in FIG. 3, and is positioned between the collection cavity 14 and the reaction cavity 17, the inlet end of the quantitative cavity 19 is in micro-fluid communication with the collection cavity 14 through a capillary tube 16, and the outlet end is in micro-fluid communication with the reaction cavity 17 through a capillary tube 16. The dosing chamber 19 is also connected to a waste chamber 13, and a solution exceeding the dosing volume is introduced into the waste chamber 13. The quantitative cavity 19 can be used for quantifying the reaction liquid, so that the quantitative requirement of detection is met.

Example 3

The difference from embodiment 2 is that, on the basis of embodiment 2, a plurality of detection areas 1B are provided, as shown in FIG. 4, and 3 detection areas are provided in this embodiment, and a plurality of quantitative cavities 19 are all in micro-fluid communication with the collection cavity 14 through the capillary 16 and communicate with the same waste liquid cavity 13. The sample nucleic acid molecules in the collection cavity 14 enter each quantitative cavity 19 to be quantified, and then enter each reaction cavity 17 and the detection cavity 18 to be respectively reacted and detected, so that one-to-many high-flux detection can be realized.

The reaction liquid injected into the reaction cavity 17 can be different, the detection medium in the detection cavity 18 can also be different, and one target gene can be detected every other reaction cavity 17, so that one sample and multiple target detections can be realized. For example, DNA of rice leaves is prepared in a sample area and then enters a plurality of reaction cavities 17, and each reaction cavity 17 detects a pathogen, so that multiple detection of rice diseases can be carried out.

Example 4

As shown in FIG. 5, the difference from example 2 is that, in addition to example 2, the nucleic acid extracting region 1A is further increased: a cleaning liquid cavity 10a and an elution liquid cavity 10b, both of which are provided with injection holes 15, wherein the cleaning liquid cavity 10a and the elution liquid cavity 10b are communicated with the collection cavity 14 through capillary tubes 16. In this embodiment, independently set up washing liquid chamber 10a and eluent chamber 10b, the washing liquid separately pours into with the eluent, can also deposit eluent or washing liquid in advance in the intracavity that corresponds, closes corresponding valve, need not to carry the strorage device of eluent and washing liquid in addition, and is more portable, during the use, only need open corresponding valve can. A sealing cover is arranged at the opening.

Example 5:

this example is different from example 1 in two places:

1. the filling hole 15 of sample cavity 11, the filling hole 15 of reaction chamber 17, the filling hole 15 department of detection chamber 18 are provided with the sealing rubber circle, the centre bore of sealing rubber circle and the 16 bore adaptations of capillary of syringe, set up like this, the capillary 16 of syringe insert the back, can with the sealing rubber circle between form relative sealed state, utilize the syringe like injecting in the sample cavity 11, provide certain pressure, be favorable to the liquid in the corresponding cavity to get into next procedure smoothly.

2. The molecular detection device is additionally provided with a box body for accommodating original pieces such as a sample processing container, an extraction detection box and the like, so that the whole device is convenient to store, store and carry. A handle is arranged on the box body. The box body is internally provided with adaptive grooves corresponding to the original elements. The sponge cushion is arranged in the groove, so that certain buffering is provided, and the damage to the molecule detection device is prevented.

Claims (10)

1. A molecular testing device adapted for in situ testing, comprising:

a sample processing container for performing cell lysis processing on a sample;

an extraction test cassette comprising a nucleic acid extraction zone and a detection zone;

a heating platform for heating the whole or the detection area part of the extraction detection box to reach the reaction temperature of nucleic acid amplification;

the detection zone includes: the device comprises a reaction cavity and a detection cavity, wherein the reaction cavity is communicated with the detection cavity in a fluid mode, and the reaction cavity is provided with an injection hole.

2. The apparatus according to claim 1, wherein the nucleic acid extraction region comprises: the sample cavity is provided with an injection hole and is communicated with the nucleic acid binding cavity in a fluid way, and the nucleic acid binding cavity is respectively communicated or closed with the waste liquid cavity and the collection cavity through a valve; an extraction carrier for combining nucleic acid molecules is arranged in the nucleic acid combining cavity.

3. The apparatus according to claim 2, wherein the nucleic acid extraction region further comprises: and the cleaning liquid cavity and the elution liquid cavity are both provided with injection holes, and the cleaning liquid cavity and the elution liquid cavity are both in fluid communication with the collection cavity.

4. The molecular testing device of claim 2, wherein the testing area further comprises: a dosing chamber having an inlet end in fluid communication with the collection chamber and an outlet end in fluid communication with the reaction chamber.

5. The device according to claim 4, wherein the detection area is provided in a plurality, and the plurality of quantitative cavities are all in fluid communication with the collection cavity.

6. The device according to claim 1, wherein the heating platform has a heating slot for inserting all or part of the extraction cartridge.

7. The device according to claim 2, wherein the extraction carrier is filter paper or a silica gel membrane.

8. The apparatus of claim 1, further comprising an extractor.

9. The apparatus of claim 1, wherein the apparatus comprises a grinding tool for grinding the sample.

10. The apparatus of claim 1, further comprising a sampling device.

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