CN210340943U - Integrated multi-linked test strip for LAMP or RT-LAMP - Google Patents
Integrated multi-linked test strip for LAMP or RT-LAMP Download PDFInfo
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Abstract
The utility model relates to an integrated multi-link test strip for LAMP or RT-LAMP, which comprises a test strip main body, wherein the test strip main body comprises an upper layer sheet, a lower layer sheet and a middle layer sheet; the upper layer sheet and the lower layer sheet are fixedly connected in a sealing manner to form a sleeve-shaped component, and one end of the sleeve-shaped component is provided with an opening; the middle layer sheet is arranged in the sleeve-shaped component, and the middle layer sheet extends out of the drawing handle towards the opening; the middle layer sheet and the sleeve-shaped component form a moving pair; the upper layer sheet is provided with a group of sample adding holes, and the lower layer sheet is provided with a group of reaction points; the middle layer sheet is provided with a group of flow holes, a group of reagent holes, a reaction zone and a group of dye holes. The material of the reaction point is FTA filter membrane or FTA card or Whatman No.1 filter paper. Adopt the utility model discloses can directly realize extraction, amplification and the demonstration result of sample nucleic acid, and can realize the detection to a plurality of target nucleic acids in the lump, break away from the dependence to the automatic instrument.
Description
Technical Field
The utility model relates to an integrated one type of many joint test paper strips for LAMP or RT-LAMP belongs to nucleic acid detection auxiliary device technical field.
Background
As far as the inventor knows, some infectious diseases with the same syndrome (such as viral hemorrhagic fever series with fever with hemorrhage, viral rash series with fever with eruption and the like) have similar main clinical manifestations, are difficult to identify and diagnose in early stage, and finally, the cause of the infectious diseases is determined by pathogen detection, and the screening and the definite diagnosis as soon as possible are crucial to control the epidemic situation. Because of the sudden and rapid spreading of the epidemic situation of the infectious disease, the rapid pathogen detection can be preferably carried out on site at the first time, and because the patient specimen or the insect-borne specimen is rolled from the site or the field and then sent to a laboratory with conditions, the distance is long, the time consumption is long, the treatment and the prevention and the control are delayed, the specimen is volatile and can cause false negative, and in addition, certain transmission hidden troubles exist. However, the existing field detection means for infectious disease pathogens are relatively thin, mainly take colloidal gold test strips, have the problems of low sensitivity, narrow detection spectrum, hook effect and the like, and often cause false negative.
Nucleic acid detection is an early, reliable, sensitive method. However, the conventional PCR and DNA probe hybridization techniques are not suitable for clinical diagnosis due to the problems of high false positive and the like; the real-time fluorescent quantitative PCR technology has high sensitivity and good reliability, is a totally enclosed reaction, but the technology needs a fluorescent quantitative PCR instrument with high price, and often needs a fluorescent labeled probe to ensure high specificity, the detection cost is higher, and the precision expensive instrument has higher requirements on quality control, operation environment and professional ability of personnel, is difficult to popularize and use in basic health departments and epidemic prevention institutions, and is also difficult to be directly used for field detection.
Compared with the detection means, the LAMP (loop-mediated isothermal amplification) or RT-LAMP (reverse transcription loop-mediated isothermal amplification) method can better screen early pathogeny on site.
The LAMP/RT-LAMP method has higher specificity and anti-interference capability, and can amplify only when the primer group is matched with 6-8 regions of the target fragment; the reaction system is stable and reliable, is still stable after being placed for 2 weeks at room temperature, is still insensitive to irrelevant and interfering fragments of original or pollution in the sample, and cannot be realized by other similar technologies; high sensitivity, fast amplification, high efficiency, and product amount up to 10 in 1 hr10The reverse transcription reaction and LAMP amplification can be carried out simultaneously, and the amplification efficiency and the detection sensitivity are about 10 to 100 times higher than those of the common RT-PCR; the result identification is relatively simple and convenient, whether the amplification is carried out or not can be judged by detecting the precipitation turbidity of the reaction tube, the color change after the reaction can be judged by directly observing the color change of a wax-sealed (high melting point) SYBR fluorescent dye at the tube cover or the tube bottom, or the color development can be greatly enhanced by directly adding a hydroxynaphthol blue dye (HNB) which does not influence the amplification efficiency into the reaction system. In a word, the LAMP/RT-LAMP method has lower requirements on sample treatment, operation technology and instruments and equipment, and is more suitable for being used in the scenes of field detection, medical institutions (such as various small and medium hospitals, community hospitals and the like), Point of care testing (POCT) and the like.
However, the application of the LAMP/RT-LAMP method to the above scenes such as field detection also has a bottleneck problem: firstly, the method needs to extract nucleic acid, the manual operation is complicated, or a full-automatic nucleic acid extractor with high price needs to be configured, and the problems of pollution and false positive are easily caused by the separation of nucleic acid extraction and detection; secondly, the method needs to prepare a reaction system, and the integration degree is not high; thirdly, related reagents of LAMP/RT-LAMP generally need to be stored in a cold chain, and are inconvenient to transport and store; fourth, at present, LAMP/RT-LAMP reaction is difficult to realize multiple reactions, generally only can detect single pathogen, has insufficient flux, and is difficult to adapt to the requirement of screening multiple pathogens of the same syndrome (such as four screening of fever with hemorrhage, infectious disease and the like). Therefore, it is necessary to develop a technical means capable of overcoming the above technical problems, so as to facilitate the application of the LAMP/RT-LAMP method in field detection and clinical diagnosis in hospitals.
The Chinese patent application with the application number of CN201610710500.3 and the application publication number of CN106337085A, which are found by retrieval, discloses a nucleic acid detection card and a use method thereof. Opening the sealing film, adding the pathogen sample into the reaction holes packaged with the nucleic acid collection test paper, automatically washing for several times by the nucleic acid purification reagent and the washing buffer solution, respectively adding the nucleic acid amplification reagent in the reagent holes into each corresponding reaction hole, carrying out nucleic acid amplification reaction under the protection of an organic phase for preventing the solution from volatilizing, and carrying out qualitative and quantitative analysis on the reaction result by using a matched optical detector.
Chinese patent application No. CN201810583631.9 and application publication No. CN108642569A disclose a nucleic acid detecting chip, comprising: a sample chamber unit disposed at an intermediate position, storing the added nucleic acid sample, and equally dividing the nucleic acid sample under rotation; a first siphon valve communicating with the sample chamber unit and introducing the nucleic acid sample into the reaction chamber unit by a siphon phenomenon; a reaction chamber unit which is provided at an edge position and mixes a nucleic acid sample with a reagent to perform a nucleic acid reaction, the reaction chamber of the reaction chamber unit being further communicated with a second siphon valve; and a reagent storage and introduction unit which is communicated with the reaction chamber unit and stores a reaction reagent, wherein after the nucleic acid sample is introduced into the reaction chamber unit, the reaction reagent is introduced into the reaction chamber unit so that the reaction of the reaction reagent and the nucleic acid sample can be detected for various blood viruses.
However, the above two technical solutions cannot be separated from expensive automatic instruments, and are difficult to adapt to the current situations of the current scene of field detection and the like.
The Chinese invention patents of patent numbers CN201410363297.8 and CN104101706B disclose a colloidal gold immunochromatographic test strip for detecting influenza A antigen and a method for detecting influenza A antigen, wherein the test strip comprises a bottom plate, and a sample pad, a conjugate pad, a nitrocellulose membrane and absorbent paper which are sequentially overlapped and stuck on the bottom plate, wherein the conjugate pad is a combination of a gold-labeled-biotin conjugate pad and a streptavidin-gold-antibody conjugate pad; in the method for detecting the influenza A antigen, a sample to be detected is dripped on the test strip, and if a red strip is displayed at the detection line (1), the sample to be detected contains the influenza A antigen.
Chinese patent application No. CN201710569444.0 and application publication No. CN107287351A disclose a Lateral flow test strip (LFD) which is based on the principle that a probe labeled with FITC is specifically hybridized with a biotin-labeled LAMP amplification product, and LFD detection does not require special equipment, and an operator only needs to immerse the product and the test strip in a buffer solution, and the LFD test strip has a control band and a detection band, both bands are colored to indicate that the detection result is positive, and only the control band is colored to indicate that the detection result is negative.
However, the above two test strip technical solutions cannot directly perform elution and amplification of nucleic acid on a test strip, and still cannot meet the requirements of scenes such as field detection.
SUMMERY OF THE UTILITY MODEL
The utility model discloses aim at: aiming at the problems in the prior art, the integrated multi-linked test strip for LAMP or RT-LAMP is provided, can directly realize the extraction, amplification and result display of sample nucleic acid, can realize the detection of a plurality of target nucleic acids together, gets rid of the dependence on an automatic instrument, is suitable for a field detection scene, and is particularly suitable for field control of infectious diseases, clinical departments of hospitals and port inspection and quarantine.
In order to achieve the above purpose, the technical scheme of the utility model is as follows:
an integrated multi-linked test strip for LAMP or RT-LAMP comprises a test strip main body, and is characterized in that the test strip main body comprises an upper layer sheet, a lower layer sheet and a middle layer sheet; the upper layer sheet and the lower layer sheet are fixedly connected in a sealing mode to form a sleeve-shaped component, and one end of the sleeve-shaped component is provided with an opening; the middle layer sheet is arranged in the sleeve-shaped member, and a drawing handle extends out of the middle layer sheet towards the opening and penetrates through the opening and extends out of the sleeve-shaped member; the middle layer sheet and the sleeve-shaped component form a sliding pair;
the upper layer sheet is provided with a group of sample adding holes, the lower layer sheet is provided with a group of reaction points, and the sample adding holes correspond to the reaction points one by one; the middle layer sheet is provided with a group of flow holes, a group of reagent holes, a reaction area and a group of dye holes, the flow holes, the reagent holes and the dye holes are in one-to-one correspondence, and the flow holes, the reagent holes and the dye holes are in one-to-one correspondence with the sample adding holes respectively;
the sample adding hole and the flow hole are respectively open holes, the reagent hole is a semi-sealed hole pre-filled with amplification reaction reagent freeze-dried powder, and the dye hole is a semi-sealed hole pre-filled with dye powder; openings at two ends of the semi-sealed hole are respectively sealed by a water permeable membrane; the reaction zone is respectively attached to the upper layer sheet and the lower layer sheet.
By adopting the structure, the test strip can be provided with a plurality of detection points to form a multi-connected test strip, when in use, the middle layer sheet can be pulled to ensure that the circulation hole, the reagent hole, the reaction area and the dye hole are sequentially positioned below the sample adding hole of the upper layer sheet and on the reaction point of the lower layer sheet, then the LAMP or RT-LAMP experimental process of adding samples and extracting nucleic acid, adding amplification reaction reagent, carrying out constant temperature amplification reaction and developing to judge the result is carried out, the detection of a plurality of target nucleic acids can be realized together, the dependence on an automatic instrument is eliminated, no additional nucleic acid extraction operation is needed, and the multi-connected test strip is particularly suitable for a field detection scene.
The utility model discloses can also adopt following preferred scheme:
preferably, the material of the reaction point is FTA filter membrane or FTA card or Whatman No.1 filter paper.
Preferably, the test strip has:
the middle layer sheet leads the flow holes to be aligned with the sample adding holes and the reaction points under the action of external force drawing, each flow hole respectively forms a sample adding cavity with the corresponding sample adding hole and the corresponding reaction point, and the sample adding state of the sample to be detected is added to the reaction points through the sample adding holes and the flow holes;
on the basis of the sample adding state and after a sample to be detected is added, and waste liquid is washed and removed, the middle layer piece enables reagent holes of the middle layer piece to be aligned to the sample adding holes and the reaction points under the action of external force drawing, each reagent hole and the corresponding sample adding hole and the corresponding reaction point form a sample adding cavity, and deionized water redissolving amplification reaction reagent freeze-dried powder is added into the reagent holes through the sample adding holes so as to add a preparation state of an amplification reaction reagent into the reaction points;
on the basis of a preparation state and after an amplification reaction reagent is added, the middle layer piece enables the reaction area to cover all reaction points under the action of external force drawing, the upper layer piece is attached to the reaction area to enable the sample adding hole to face the lower end opening of the middle layer piece to be kept closed, the lower layer piece is attached to the reaction area to enable the reaction points to be located in a closed space, and the area where the sample adding hole of the test strip main body is located is placed in a constant temperature environment to be in an amplification reaction state of reaction;
on the basis of the amplification reaction state and after the amplification reaction is finished, the middle layer piece enables the dye holes to be aligned to the sample adding holes and the reaction points under the action of external force drawing, each dye hole and the corresponding sample adding hole and the corresponding reaction point form a sample adding cavity, deionized water is added into the dye holes through the sample adding holes to dissolve dye powder so as to add the dye into the reaction points, and the dye powder is developed so as to judge the result display state of the result.
By adopting the preferable scheme, the LAMP or RT-LAMP experiment process can be smoothly implemented on the test strip through more definite states.
Preferably, the gap between the middle layer sheet and the sleeve-like member is filled with grease, and the grease is located around the flow-through holes, the reagent holes, the reaction zones, the dye holes, the sample addition holes, and the reaction sites.
Preferably, the upper layer sheet, the middle layer sheet and the lower layer sheet are respectively made of polymethyl methacrylate; and a liquid silica gel layer is filled in a gap between the middle layer sheet and the sleeve-shaped member, the liquid silica gel layer is fixedly connected with the sleeve-shaped member, and the liquid silica gel layer is positioned around the circulation hole, the reagent hole, the reaction zone, the dye hole, the sample adding hole and the reaction point.
By adopting any one of the two optimal schemes, better sealing effect can be realized, the reliability of detection can be better ensured, and aerosol pollution possibly caused by amplification reaction can be avoided as much as possible.
Preferably, a first magnetic sheet is arranged in the upper layer sheet, a second magnetic sheet is arranged in the lower layer sheet, and the first magnetic sheet and the second magnetic sheet attract each other by magnetic force; the middle layer sheet is made of a ferrite magnetic film.
By adopting the optimal scheme, the upper layer sheet and the lower layer sheet can clamp the middle layer sheet by means of magnetic attraction, the material of the middle layer sheet can further ensure the clamping effect, better sealing effect is realized, the detection reliability is better ensured, and aerosol pollution possibly brought by amplification reaction is avoided as much as possible.
Preferably, the drawing handle of the middle layer sheet is provided with a group of drawing indication lines, and the drawing indication lines comprise a preparation state indication line, an amplification reaction state indication line and a result display state indication line. When the test strip is in a sample adding state, the preparation state indicating line, the amplification reaction state indicating line and the result display state indicating line are all positioned in the opening of the sleeve-shaped component; when the test strip is in a ready state, the ready state indicating line is flush with the outer edge of the opening of the sleeve-shaped member, and the amplification reaction state indicating line and the result display state indicating line are both positioned in the opening of the sleeve-shaped member; when the test strip is in an amplification reaction state, the preparation state indicating line is positioned outside the opening of the sleeve-shaped member, the amplification reaction state indicating line is flush with the outer edge of the opening of the sleeve-shaped member, and the result shows that the state indicating line is positioned inside the opening of the sleeve-shaped member; when the test strip is in a result display state, the preliminary state indicating line and the amplification reaction state indicating line are positioned outside the opening of the sleeve-shaped member, and the result display state indicating line is flush with the outer edge of the opening of the sleeve-shaped member.
By adopting the preferred scheme, the current state of the test strip can be indicated by drawing the indicating line, so that the test strip is easier to use.
Preferably, the withdrawal handle of the middle layer sheet is matched with the size of the opening of the sleeve-shaped member, and the withdrawal handle is embedded in the opening.
By adopting the preferable scheme, the test strip has better sealing performance, and the whole middle layer sheet is prevented from being pulled out.
Preferably, a liquid silica gel layer is arranged on the side surface of the reaction zone close to the lower layer sheet, and a liquid silica gel layer is also arranged at the part of the lower layer sheet around the reaction point; and when the reaction point is positioned in the coverage area of the reaction area, the liquid silica gel layer of the reaction area is attached to the liquid silica gel layer of the lower layer sheet.
By adopting the preferable scheme, when the reaction point is positioned in the coverage area of the reaction area, the reaction point can be more effectively sealed, so that after the whole test strip or the reaction area is placed in a constant-temperature environment (such as a water bath, a constant-temperature device and the like), the reaction point can carry out amplification reaction in a sealed state, the detection reliability is better ensured, and aerosol pollution possibly caused by the amplification reaction is avoided as much as possible.
Adopt the utility model discloses afterwards, can be directly realize nucleic acid extraction through the mode that adds the reagent on the test paper strip, need not prepare whole reaction system simultaneously, can directly realize extraction, amplification and the demonstration result of sample nucleic acid, realize the detection to a plurality of target nucleic acids in the lump, break away from the dependence to the automatic instrument, be applicable to the witnessed inspections scene.
Drawings
The present invention will be further described with reference to the accompanying drawings.
Fig. 1 is a schematic view of a main structure of an embodiment of the present invention.
Fig. 2 is a schematic view of a sleeve member according to an embodiment of the present invention.
Fig. 3 is a partially enlarged schematic view of fig. 2.
Fig. 4 is a schematic view of a front view of an embodiment of the present invention.
Fig. 5 is a schematic view of a middle layer sheet according to an embodiment of the present invention.
Fig. 6 is a schematic view of a lower laminate according to an embodiment of the present invention.
Fig. 7 is a schematic sectional view of a-a in fig. 4 when the test strip of the embodiment of the present invention is in the sample adding state.
Fig. 8 is a schematic sectional view of a-a in fig. 4 when the test strip of the embodiment of the present invention is in an amplification reaction state.
Detailed Description
Examples
As shown in fig. 1 to 8, the integrated multiplex test strip for LAMP or RT-LAMP of the present embodiment includes a test strip body 01, the test strip body 01 including an upper layer sheet 11, a lower layer sheet 13, and a middle layer sheet 12; the upper layer sheet 11 and the lower layer sheet 13 are fixedly connected in a sealing way to form a sleeve-shaped component 14, and one end of the sleeve-shaped component 14 is provided with an opening 15; the middle layer sheet 12 is arranged in the sleeve-shaped member 14, the middle layer sheet 12 extends out of a drawing handle 16 towards the opening 15, and the drawing handle 16 passes through the opening 15 and extends out of the sleeve-shaped member 14; the middle layer sheet 12 and the sleeve-shaped component 14 form a moving pair; the pull handle 16 of the middle layer 12 is sized to fit the opening 15 of the sleeve member 14, and the pull handle 16 is embedded within the opening 15.
The upper layer sheet 11 is provided with a group of sample adding holes 21, the lower layer sheet 13 is provided with a group of reaction points 28 (the material of the reaction points 28 is an FTA filter membrane or an FTA card or Whatman No.1 filter paper, the diameter of the reaction points 28 can be set to be about 5 mm), and the sample adding holes 21 are in one-to-one correspondence with the reaction points 28; the middle layer sheet 12 is provided with a group of flow holes 23, a group of reagent holes 25, a reaction area 24 and a group of dye holes 26, wherein the flow holes 23, the reagent holes 25 and the dye holes 26 are in one-to-one correspondence, and the flow holes 23, the reagent holes 25 and the dye holes 26 are in one-to-one correspondence with the sample adding holes 21 respectively.
The sample adding hole 21 and the flow hole 23 are respectively open holes, the reagent hole 25 is a semi-sealed hole pre-filled with amplification reaction reagent freeze-dried powder, and the dye hole 26 is a semi-sealed hole pre-filled with dye powder; openings at two ends of the semi-sealed hole are respectively sealed by a water permeable membrane; the reaction zone 24 is attached to the upper sheet 11 and the lower sheet 13, respectively.
To ensure the sealing performance of the test strip, one of the following two structures (as shown in fig. 7) can be adopted:
(1) the gap between the middle layer sheet 12 and the sleeve member 14 is filled with grease 41, and the grease 41 is located around the flow hole 23, the reagent hole 25, the reaction region 24, the dye hole 26, the loading hole 21, and the reaction site 28.
Dupont may be used as the grease 41
lvp high vacuum grease 41. The grease 41 can form a lubricating film between the middle sheet 12 and the sleeve member 14 and serves as a sealing function.
(2) The upper layer sheet 11, the middle layer sheet 12 and the lower layer sheet 13 are respectively made of polymethyl methacrylate; the gap between the middle layer sheet 12 and the sleeve-shaped member 14 is filled with a liquid silica gel layer 42, the liquid silica gel layer 42 is fixedly connected with the sleeve-shaped member 14, and the liquid silica gel layer 42 is positioned around the flow holes 23, the reagent holes 25, the reaction zone 24, the dye holes 26, the sample adding holes 21 and the reaction points 28.
Note: the liquid silicone layer 42 is a solid layer made of liquid silicone, and is not a liquid layer.
In order to further ensure the sealing performance of the test strip, as shown in fig. 7, a first magnetic sheet 43 is disposed in the upper layer sheet 11, a second magnetic sheet 44 is disposed in the lower layer sheet 13, and the first magnetic sheet 43 and the second magnetic sheet 44 attract each other by magnetic force; the material of the intermediate layer sheet 12 is a ferrite magnetic thin film.
In order to further ensure the sealing performance of the reaction site of the test strip during the amplification reaction, as shown in fig. 6 and 8, a liquid silica gel layer 45 is disposed on the side surface of the reaction region 24 close to the lower layer sheet 13, and a liquid silica gel layer 46 is also disposed on the lower layer sheet 13 around the reaction site 28; when the reaction site 28 is located within the coverage area of the reaction zone 24, the liquid silica gel layer 45 of the reaction zone 24 is attached to the liquid silica gel layer 46 of the lower sheet 13.
Through tests, the test strip of the embodiment can meet the requirement of more than 1 hour of incubation at 65 ℃.
The integrated test strip of the embodiment has the following specific states:
(1) the middle layer sheet 12 is pulled by an external force to align the flow holes 23 with the sample addition holes 21 and the reaction sites 28, each flow hole 23 forms a sample addition chamber with the corresponding sample addition hole 21 and the corresponding reaction site 28, and the sample addition state of the sample to be tested is added to the reaction sites 28 through the sample addition holes 21 and the flow holes 23. (Note: the sample to be examined is blood, a bacterial culture solution, etc.)
(2) On the basis of the sample adding state, after a sample to be detected is added, and waste liquid is washed and removed, the middle layer piece 12 enables the reagent holes 25 to be aligned to the sample adding holes 21 and the reaction points 28 under the action of external force drawing, each reagent hole 25 forms a sample adding cavity with the corresponding sample adding hole 21 and the corresponding reaction point 28, and deionized water redissolving amplification reaction reagent freeze-dried powder is added into the reagent holes 25 through the sample adding holes 21 so as to add the preparation state of the amplification reaction reagent into the reaction points 28. (note: according to the normal test steps, after adding the sample to be detected, waiting for a period of time, adding FTA extracting solution and TE buffer solution, washing once respectively, and sucking all waste liquid by using a pipettor, wherein the freeze-dried powder of the amplification reaction reagent comprises an LAMP detection reagent and an amplification primer.)
(3) On the basis of a preparation state and after adding an amplification reaction reagent, the middle layer sheet 12 enables the reaction area 24 to cover all reaction points 28 under the action of external force drawing, the upper layer sheet 11 is attached to the reaction area 24 to enable the sample adding holes 21 to face the lower end opening 15 of the middle layer sheet 12 to be kept closed, the lower layer sheet 13 is attached to the reaction area 24 to enable the reaction points 28 to be located in a closed space, and the area where the sample adding holes 21 of the test strip main body 01 are located is placed in a constant temperature environment to be in an amplification reaction state of reaction.
(4) On the basis of the amplification reaction state and after the amplification reaction is completed, the middle layer 12 makes the dye holes 26 align with the sample adding holes 21 and the reaction points 28 under the action of external force drawing, each dye hole 26 forms a sample adding cavity with the corresponding sample adding hole 21 and the corresponding reaction point 28, deionized water is added into the dye holes 26 through the sample adding holes 21 to dissolve dye powder so as to add the dye into the reaction points 28, and the dye is developed so as to determine the result display state of the result. (Note: the dye powder was SYBR green powder.)
As shown in FIG. 5, the withdrawal handle 16 of the middle layer sheet 12 is provided with a set of withdrawal indicating lines including a preparatory status indicating line 30, an amplification reaction status indicating line 31, and a result display status indicating line 32.
When the test strip is in the sample loading state, the preparation state indicating line 30, the amplification reaction state indicating line 31, and the result display state indicating line 32 are all located within the opening 15 of the sleeve-shaped member 14; when the strip is in the ready state, the ready state indicating line 30 is flush with the outer edge of the opening 15 of the sleeve-shaped member 14, and the amplification reaction state indicating line 31 and the result display state indicating line 32 are both located inside the opening 15 of the sleeve-shaped member 14; when the strip is in the amplification reaction state, the ready state indicating line 30 is located outside the opening 15 of the sleeve-shaped member 14, the amplification reaction state indicating line 31 is flush with the outer edge of the opening 15 of the sleeve-shaped member 14, and as a result, the display state indicating line 32 is located inside the opening 15 of the sleeve-shaped member 14; when the strip is in the result display state, the preliminary state indicating line 30 and the amplification reaction state indicating line 31 are located outside the opening 15 of the sheath member 14, and the result display state indicating line 32 is flush with the outer edge of the opening 15 of the sheath member 14.
When the test strip is used specifically, the drawing indication lines can be combined, the middle layer sheet 12 is drawn to enable the circulation holes 23, the reagent holes 25, the reaction area 24 and the dye holes 26 to be aligned with or cover the sample adding holes 21 of the upper layer sheet 11 and the reaction points 28 of the lower layer sheet 13 in sequence, and LAMP or RT-LAMP experimental processes of adding a sample to be detected, adding eluent to elute and extract nucleic acid, adding an amplification reaction reagent, placing in a constant temperature environment to perform constant temperature amplification reaction, and adding dye to develop color so as to judge results are correspondingly implemented, so that detection of a plurality of target nucleic acids is finally realized.
Wherein, a water bath kettle or small-sized constant temperature equipment can be adopted during the constant temperature amplification reaction; before the test strip is put into a constant temperature environment, the test strip is pressed by external force, so that the liquid silica gel layer 45 of the reaction area 24 can be better attached to the liquid silica gel layer 46 of the lower layer sheet 13; when the test strip is placed in a constant temperature environment, the whole test strip is usually placed, and if conditions allow, only the reaction area 24 may be placed in a constant temperature environment for the amplification reaction.
In addition to the above embodiments, the present invention may have other embodiments. All the technical solutions formed by adopting equivalent substitutions or equivalent transformations fall within the protection scope claimed by the present invention.
Claims (10)
1. An integrated multi-linked test strip for LAMP or RT-LAMP comprises a test strip main body, and is characterized in that the test strip main body comprises an upper layer sheet, a lower layer sheet and a middle layer sheet; the upper layer sheet and the lower layer sheet are fixedly connected in a sealing mode to form a sleeve-shaped component, and one end of the sleeve-shaped component is provided with an opening; the middle layer sheet is arranged in the sleeve-shaped member, and a drawing handle extends out of the middle layer sheet towards the opening and penetrates through the opening and extends out of the sleeve-shaped member; the middle layer sheet and the sleeve-shaped component form a sliding pair;
the upper layer sheet is provided with a group of sample adding holes, the lower layer sheet is provided with a group of reaction points, and the sample adding holes correspond to the reaction points one by one; the middle layer sheet is provided with a group of flow holes, a group of reagent holes, a reaction area and a group of dye holes, the flow holes, the reagent holes and the dye holes are in one-to-one correspondence, and the flow holes, the reagent holes and the dye holes are in one-to-one correspondence with the sample adding holes respectively;
the sample adding hole and the flow hole are respectively open holes, the reagent hole is a semi-sealed hole pre-filled with amplification reaction reagent freeze-dried powder, and the dye hole is a semi-sealed hole pre-filled with dye powder; openings at two ends of the semi-sealed hole are respectively sealed by a water permeable membrane; the reaction zone is respectively attached to the upper layer sheet and the lower layer sheet.
2. The integrated multiplex test strip for LAMP or RT-LAMP according to claim 1, wherein the reaction sites are made of FTA filter membranes, FTA cards or Whatman No.1 filter paper.
3. The integrated multiplex test strip for LAMP or RT-LAMP according to claim 1, characterized in that it has:
the middle layer sheet leads the flow holes to be aligned with the sample adding holes and the reaction points under the action of external force drawing, each flow hole respectively forms a sample adding cavity with the corresponding sample adding hole and the corresponding reaction point, and the sample adding state of the sample to be detected is added to the reaction points through the sample adding holes and the flow holes;
on the basis of the sample adding state and after a sample to be detected is added, and waste liquid is washed and removed, the middle layer piece enables reagent holes of the middle layer piece to be aligned to the sample adding holes and the reaction points under the action of external force drawing, each reagent hole and the corresponding sample adding hole and the corresponding reaction point form a sample adding cavity, and deionized water redissolving amplification reaction reagent freeze-dried powder is added into the reagent holes through the sample adding holes so as to add a preparation state of an amplification reaction reagent into the reaction points;
on the basis of a preparation state and after an amplification reaction reagent is added, the middle layer piece enables the reaction area to cover all reaction points under the action of external force drawing, the upper layer piece is attached to the reaction area to enable the sample adding hole to face the lower end opening of the middle layer piece to be kept closed, the lower layer piece is attached to the reaction area to enable the reaction points to be located in a closed space, and the area where the sample adding hole of the test strip main body is located is placed in a constant temperature environment to be in an amplification reaction state of reaction;
on the basis of the amplification reaction state and after the amplification reaction is finished, the middle layer piece enables the dye holes to be aligned to the sample adding holes and the reaction points under the action of external force drawing, each dye hole and the corresponding sample adding hole and the corresponding reaction point form a sample adding cavity, deionized water is added into the dye holes through the sample adding holes to dissolve dye powder so as to add the dye into the reaction points, and the dye powder is developed so as to judge the result display state of the result.
4. The integrated multiplex test strip for LAMP or RT-LAMP according to claim 1, wherein the gap between the middle layer sheet and the sleeve-like member is filled with grease, and the grease is located around the flow-through hole, the reagent hole, the reaction zone, the dye hole, the sample addition hole, and the reaction site.
5. The integrated multi-linked test strip for LAMP or RT-LAMP according to claim 1, wherein the upper layer sheet, the middle layer sheet and the lower layer sheet are made of polymethyl methacrylate respectively; and a liquid silica gel layer is filled in a gap between the middle layer sheet and the sleeve-shaped member, the liquid silica gel layer is fixedly connected with the sleeve-shaped member, and the liquid silica gel layer is positioned around the circulation hole, the reagent hole, the reaction zone, the dye hole, the sample adding hole and the reaction point.
6. The integrated multiplex test strip for LAMP or RT-LAMP according to claim 1, wherein a first magnetic sheet is provided in the upper layer sheet, a second magnetic sheet is provided in the lower layer sheet, and the first magnetic sheet and the second magnetic sheet attract each other by magnetic force; the middle layer sheet is made of a ferrite magnetic film.
7. The integrated multi-linked test strip for LAMP or RT-LAMP according to claim 1, wherein the drawing handle of the middle layer sheet is provided with a set of drawing indication lines, and the drawing indication lines comprise a preparation state indication line, an amplification reaction state indication line and a result display state indication line.
8. The integrated multiplex test strip for LAMP or RT-LAMP according to claim 7, wherein when the test strip is in the sample loading state, the ready state indicating line, the amplification reaction state indicating line, and the result display state indicating line are all located within the opening of the sleeve-like member; when the test strip is in a ready state, the ready state indicating line is flush with the outer edge of the opening of the sleeve-shaped member, and the amplification reaction state indicating line and the result display state indicating line are both positioned in the opening of the sleeve-shaped member; when the test strip is in an amplification reaction state, the preparation state indicating line is positioned outside the opening of the sleeve-shaped member, the amplification reaction state indicating line is flush with the outer edge of the opening of the sleeve-shaped member, and the result shows that the state indicating line is positioned inside the opening of the sleeve-shaped member; when the test strip is in a result display state, the preliminary state indicating line and the amplification reaction state indicating line are positioned outside the opening of the sleeve-shaped member, and the result display state indicating line is flush with the outer edge of the opening of the sleeve-shaped member.
9. The integrated multi-linked test strip for LAMP or RT-LAMP according to any one of claims 1 to 8, wherein the drawing handle of the middle layer sheet is matched with the size of the opening of the sleeve-shaped member, and the drawing handle is embedded in the opening.
10. The integrated multi-link test strip for LAMP or RT-LAMP according to any one of claims 1 to 8, wherein a liquid silica gel layer is arranged on the side of the reaction zone close to the lower layer sheet, and a liquid silica gel layer is also arranged on the part of the lower layer sheet around the reaction point; and when the reaction point is positioned in the coverage area of the reaction area, the liquid silica gel layer of the reaction area is attached to the liquid silica gel layer of the lower layer sheet.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921060673.0U CN210340943U (en) | 2019-07-08 | 2019-07-08 | Integrated multi-linked test strip for LAMP or RT-LAMP |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201921060673.0U CN210340943U (en) | 2019-07-08 | 2019-07-08 | Integrated multi-linked test strip for LAMP or RT-LAMP |
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| CN210340943U true CN210340943U (en) | 2020-04-17 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110218769A (en) * | 2019-07-08 | 2019-09-10 | 中国人民解放军东部战区疾病预防控制中心 | The multi-joint test strips of integration integral type for LAMP or RT-LAMP |
-
2019
- 2019-07-08 CN CN201921060673.0U patent/CN210340943U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110218769A (en) * | 2019-07-08 | 2019-09-10 | 中国人民解放军东部战区疾病预防控制中心 | The multi-joint test strips of integration integral type for LAMP or RT-LAMP |
| CN110218769B (en) * | 2019-07-08 | 2024-04-19 | 中国人民解放军东部战区疾病预防控制中心 | Integrated multi-linked test strip for LAMP or RT-LAMP |
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