Technical Field
The present invention relates to a biosensor chip that is used in the detection of bacteria, viruses, and so forth that can cause a variety of infectious diseases, and to a biosensor device equipped with this chip.
Description of the Related Art
In the past, biosensor chips of this type have been used in devices that perform nucleic acid amplification reaction, for example, in the analysis of objects included in a specimen.
A conventional biosensor chip internally comprised a main body case having a diluent chamber and a measurement chamber connected via a first channel to this diluent chamber, an inlet provided to a portion of this main body case corresponding to the diluent chamber, and a sealing member for sealing this inlet. The measurement chamber had a branching chamber component that was connected to the diluent chamber via the first channel, and a plurality of individual measurement components connected to this branching chamber component via second channels. Individual reagents were provided to the individual measurement components (see Japanese Unexamined Patent Application Publication (Translation of PCT Application) No. 2010-519892, for example).
With the prior art discussed above, an example of biological information is the identification of the presence or type of a virus. In the case of the common cold, for example, the strain (that is, the form of the virus) changes every year. To detect that virus, it is necessary to house individual reagents for detecting the virus in individual measurement components, to house a different individual reagent for each individual measurement component, and to combine a plurality of types of individual reagents.
However, there are hundreds of individual reagents corresponding to viruses that must be combined in a plurality of types for the purpose of detection, and these combinations have to be taken into account and numerous biosensor chips prepared. Accordingly, the management of so many biosensor chips in which individual reagents of different types are combined entailed a tremendous expense, and this drove up the cost.
Moreover, with a conventional configuration, when individual reagents of different types were applied and dried under the same environment, there was the risk that cross contamination between the individual reagents would occur, and this could lead to decreased measurement accuracy.
In view of this, it is an object of the present invention to prevent a decrease in measurement accuracy caused by contamination, and to thereby lower the cost.
The biosensor chip pertaining to the first invention is a biosensor chip that is placed in a biosensor device and is rotated while a specimen is measured, the biosensor chip comprising a main body, a holding chamber, a dispensing chamber, a plurality of quantification chambers, and a plurality of measurement chambers. The main body has an inlet into which a biochemical analysis specimen is poured. The holding chamber holds the poured specimen inside the main body. The dispensing chamber is connected to the holding chamber via a first channel and dispenses the specimen. The plurality of quantification chambers are connected to the dispensing chamber, hold a specific amount of dispensed specimen, and are disposed at positions located away from the rotational center of the rotary motion according to the distance from the first channel. The plurality of measurement chambers are connected to the quantification chambers via a second channel and react the specimen with a biochemical analysis reagent.
In a configuration in which this biosensor chip is placed in a biosensor device and rotated, of the plurality of quantification chambers, the farther away a quantification chamber is disposed from the first channel, the farther it is disposed away from the rotational center of the rotary motion. Thus, the quantification chambers empty of specimen starting from the side closest to the first channel, so specimen is successively supplied to the adjacent quantification chamber disposed downstream in the rotational direction.
Consequently, a specific amount of specimen can be introduced into the plurality of quantification chambers, and the amount of specimen held in the quantification chambers can be kept from becoming inconsistent, which happens when specimen flows backward from an adjacent quantification chamber.
Also, with a chip configuration in which an overflow chamber is provided on the downstream side of the plurality of quantification chambers in the rotational direction, if waves should be produced in the specimen in the overflow chamber as a result of fluctuation in the rotary speed of the rotary motion, backflow of the specimen in the overflow chamber can be prevented in the same way as when lower waves cannot quite reach a higher bank. As a result, the various quantification chambers can be kept in a state in which the specified amount of specimen has been introduced, so the proper analysis can be performed, using a consistent amount of specimen, in the measurement chambers into which the specimen is supplied from these quantification chambers.
The biosensor chip pertaining to an embodiment the present invention will now be described through reference to the drawings.
Embodiment 1
Biosensor Device
As shown in
As shown in
The rotary tray 7 is connected at its center part to a shaft connected to a rotation mechanism 8, and is rotated by the rotation mechanism 8.
A temperature sensor 10 is disposed at the top inside the space 6.
The temperature sensor 10 is provided in order to monitor the temperature inside the space 6, and the temperature in the space 6 can be adjusted to a specific temperature by switching on or off a heater 11 disposed in the space 6, on the basis of the temperature sensed by the temperature sensor 10.
An optical sensor 12 is then disposed on the bottom face of the rotary tray 7, opposite the lower face of the biosensor chip 1, in the space 6.
The optical sensor 12 senses an influenza specimen on the biosensor chip 1.
Also disposed in the interior of the device main body 2 are a computer 13 that computes values sensed by the optical sensor 12, and a controller 14 that controls the biosensor device.
With the above configuration, in this embodiment, a plurality of individual specimens that react with a plurality of types of virus are housed in the biosensor chip 1 in order to sense the type of influenza specimen.
Biosensor Chip 1
As shown in
The cover 15 has an inlet 18 for introducing a specimen at one corner of the substantially square shape.
The inlet 18 is sealed off by a sealing member 19. As shown in
A diluent is housed ahead of time in the diluent chamber 20. In the state in
Therefore, when an influenza specimen is poured out of the inlet 18, first this seal is peeled off, and then the influenza specimen is poured from the inlet 18 into the diluent chamber 20. Consequently, the influenza specimen and the diluent are mixed together, and a diluted specimen of a specific concentration is held in the diluent chamber 20.
As shown in
The measurement chamber 22 has a branched chamber 23 that is linked via the first channel 21 to the diluent chamber 20, and a plurality of individual measurement chambers 25 that are connected via second channels 24 to the branched chamber 23.
As shown in
The mounting of the common reagent 28 will now be described in greater detail through reference to
First, as shown in
The film 16a constitutes the individual measurement chambers 25 along with the base 16.
As shown in
The common reagent 28 is essential to the detection of bacteria and viruses, and performs the job of amplifying a specimen that is present in the diluted specimen. Therefore, individual reagents 29 that react individually with the bacterium or virus to be detected needs to be separately held in the individual measurement chambers 25.
In view of this, in this embodiment, as shown in
As shown in
The liquid individual reagents 29 are dropped into the interior of the recesses 31 and then dried, which fixes the individual reagents 29. The recesses 31 are substantially bowl shaped, and the surface tension of the liquid causes the individual reagents 29 to dry and be fixed in a state in which they are uniformly spread out over the inner peripheral face of the recesses 31.
As shown in
That is, individual reagents 29 of different types can be fixed to the caps 27 mounted to adjacent individual measurement chambers 25, and measurement performed. This makes it easy to sense the strain of an influenza virus, for example.
As shown in
Incidentally, a specimen collected from a patient will undergo a favorable reaction with the individual reagent 29 at around 60° C. If the temperature were considerably lower than 60° C., the reaction would take a long time, but if the temperature were considerably higher than 60° C., the specimen itself would be destroyed and there would be no reaction. Accordingly, the diluted specimen must flow into the individual measurement chambers 25 while the interior of the individual measurement chambers 25 is held at 60° C.
In view of this, with the biosensor chip 1 in this embodiment a heat-fusible sealing material that fills in the space inside the second channels 24 is provided.
The heat-fusible sealing material used here is one that will melt at approximately 40° C. Consequently, in the course of heating the individual measurement chambers 25 of the biosensor chip 1 to 60° C., the heat-fusible sealing material of the second channels 24 begins to melt, and spaces in which the diluted specimen moves are formed inside the second channels 24.
Consequently, the diluted specimen held in the branched chamber 23 can flow into the individual measurement chambers 25 under the centrifugal force produced by the biosensor chip 1 during the rotation of the rotary tray 7. As a result, the spaces inside the individual measurement chambers 25 can be maintained at about 60° C.
If the introduction of the heat-fusible sealing material into the second channels 24 is performed while using a dispenser, for example, and monitoring through the reagent mounting holes 26 in a state in which the caps 27 have not yet been mounted to the reagent mounting holes 26 in
With the configuration of the biosensor chip 1 in this embodiment, a diluted specimen is introduced into the individual measurement chambers 25 to which the individual reagents 29 have been mounted, and is subjected to an amplification reaction. This amplification reaction is detected by the optical sensor 12, which confirms which specimen is reacting with the individual reagents 29. This result is displayed on the display component 5.
With this embodiment, as discussed above, the individual reagents 29 necessary for detection are provided on the faces of the caps 27 exposed inside the individual measurement chambers 25. A configuration is employed that allows the caps 27 to be mounted to the reagent mounting holes 26 of the individual measurement chambers 25.
Consequently, the main body case 17 may be managed as a shared part, and the caps 27 may be appropriately selected and mounted according to the individual applications of the virus or the like to be detected. As a result, production costs, management costs, and other such costs can be greatly reduced.
Also, since the caps 27 are smaller than the main body case 17, even when a plurality of caps 27 are managed, they can be simply managed in a small amount of space. This also affords a cost reduction.
The biosensor chip 1 is also such that the caps 27 holding the individual reagents 29 can be mounted in the reagent mounting holes 26 of the individual measurement chambers 25. Accordingly, since the individual reagents 29 are held in the small caps 27 that afford easy environment management such as cleanliness ahead of time, contamination of the individual reagents 29 can be prevented. As a result, this avoids the adverse effects on measurement accuracy that would otherwise be caused by contamination of the individual reagents 29, and allows for more accurate measurement.
When different types of the individual reagents 29 are applied and dried under the same environment, there is the risk of contamination between the individual reagents 29.
In this embodiment, the individual reagents 29 are housed on the caps 27 apart from the main body case 17. Thus, the individual reagents 29 can be applied and dried in isolated spaces, and this prevents contamination between the individual reagents 29.
Also, even if it is necessary to house the common reagent 28 and the individual reagents 29 in a separated state in the individual measurement chambers 25 in order to avoid deactivation of the reagents, in this embodiment reagent deactivation can be easily prevented by housing the individual reagents 29 in the caps 27, and the common reagent 28 in the individual measurement chambers 25 of the main body case.
The film 16a that seals the individual measurement chambers 25 is generally formed from a plastic film or the like, and is bonded to the main body case 17 by heat sealing or another such means. Accordingly, if a reagent has low heat resistance, there is the risk that it will end up being deactivated by this heat.
In this embodiment, since there are reagent mounting holes 26 that communicate with the respective individual measurement chambers 25, the film 16a can be bonded to the main body case 17 by heat sealing, and the film 16a can then be coated with the common reagent 28 through the reagent mounting holes 26. This effectively prevents deactivation of the common reagent 28 as well.
Embodiment 2
In this embodiment, those components that are shared with Embodiment 1 above will be numbered the same, and these components will not be described in detail again.
In this embodiment, just as in Embodiment 1 above, the biosensor chip 1 into which influenza specimens have been introduced is inserted into the biosensor device shown in
That is, the biosensor chip 1 is placed inside the device main body 2 of the biosensor device, and the result of biochemical analysis is displayed on the display component 5.
As shown in
The chip main body 106 has in its interior a holding chamber 112 (discussed below), a dispensing chamber 114, quantification chambers 115, measurement chambers 117, and so forth (see
The lower cover 107 is attached to the lower face of the chip main body 106.
The inlet 108 is attached to the upper face of the chip main body 106, and specimens used for biochemical analysis are introduced into this inlet.
The inlet 108 is formed in the upper cover 109.
The accessory cover 110 is attached to the upper face of the upper cover 109.
The sealing member 111 seals the inlet 108.
The configuration of this chip main body 106 will now be described in detail.
As shown in
The holding chamber 112 holds the introduced specimen.
The dispensing chamber 114 is connected via a first channel 113 to the holding chamber 112, and dispenses specimens.
The quantification chambers 115 are connected to the dispensing chamber 114, and hold the dispensed specimens.
The measurement chambers 117 are connected to the respective quantification chambers 115 via second channels 116, and react the specimens with the biochemical analysis reagents.
The reagents used in this embodiment are applied as a coating to the interior of the measurement chambers 117 ahead of time.
As shown in
The overflow chamber 118 holds any specimen that has overflowed from the quantification chambers 115.
In this embodiment, when the biosensor chip 1 undergoes rotary motion after being placed in the biosensor device, of the plurality of quantification chambers 115, the quantification chambers 115 located away from the first channel 113 are disposed at positions farther away from the rotational center of the rotary motion than the quantification chambers 115 located closer to the first channel 113.
More precisely, as shown in
For example, as shown in
As shown in
The above-mentioned radial direction lengths of the first and second ends refer to the lengths in the radial direction of a fan shape whose center is the rotational center of the chip main body 106.
Consequently, specimens are successively supplied to the quantification chambers 115 disposed on the downstream side in the rotational direction as they are emptied of specimens starting from the quantification chambers 115 closest to the first channel 113 side.
Consequently, the specified amount of specimen can be introduced into the quantification chambers 115, so the proper analysis can be carried out.
The overflow chamber 118 is provided at the farthest downstream point in the rotational direction of the quantification chambers 115. Therefore, backflow of the specimen from the overflow chamber 118 to the quantification chamber 115 side can be prevented during rotary motion. As a result, the amount of specimen held in the quantification chambers 115 can be stabilized, and the proper analysis can be carried out.
That is, in this embodiment, specimens are supplied from the holding chamber 112, through the dispensing chamber 114 and the quantification chambers 115, to the measurement chambers 117 by rotating the chip main body 106 that has been placed in the biosensor device.
Here, in order to promote a reaction between the reagents and the specimens in the measurement chambers 117, the rotational speed of the chip main body 106 is sometimes changed to agitate the specimens and the reagents. This change in the rotational speed can create waves in the specimen in the overflow chamber 118, and there is the risk that these will flow back into the quantification chambers 115.
In view of this, in this embodiment the quantification chambers 115 are disposed at locations that are distant from the rotational center of the rotary motion of the chip main body 106 according to the distance from the first channel 113.
Consequently, even if waves should be generated in the specimen in the overflow chamber 118 by a change in rotational speed, the specimen can be prevented from flowing backward in the same way as when lower waves cannot quite reach a higher bank. As a result, the various quantification chambers 115 can be kept in a state in which the specified amount of specimen has been introduced. Accordingly, the amount of specimen held in the measurement chambers 117 to which specimens are supplied from the quantification chambers 115 can be stabilized, so the proper analysis can be performed.
The overflow chamber 118 is disposed so that the distance d5 from the rotational center of the chip main body 106 to the distance d4 of the adjacent measurement chamber 115 will be d4<d5. This allows any specimen that has overflowed from the quantification chambers 115 to be stably introduced into the overflow chamber 118.
Furthermore, once specimen has flowed into the overflow chamber 118, it is prevented from flowing back to the measurement chambers 115 located on the upstream side in the rotational direction.
As a result, the specified amount of specimen can be stably held in the quantification chambers 115, and the proper analysis can be conducted.
Embodiment 3
In this embodiment, those components that are shared with Embodiment 1 above will be numbered the same, and these components will not be described in detail again.
The configuration shown in
With the above configuration, in this embodiment a biosensor chip 1 that detects an influenza virus and also identifies the strain will be described in detail as an example of biological information about a specimen collected from a patient's nostrils.
As shown in
The main body case 219 has an inlet 220 at the portion corresponding to the diluent chamber 216. The inlet 220 is sealed from the outside of the main body case 219 by a sealing member 221.
As shown in
A diluent for diluting a specimen is sealed in the interior of the diluent chamber 216. As shown in
As shown in
As shown in
Then, as shown in
The specimen here is moved in and out of the diluent chamber 216 through the inlet 220 from and into the pipette, which mixes the introduced specimen with the diluent and results in a mixture of consistent concentration.
Once the introduction of the specimen into the biosensor chip 1 is thus complete, the user seals off the inlet 220 with the sealing member 221.
As shown in
Accordingly, when the sealing member 221 is pushed into the inlet 220, the sealing member 221 undergoes elastic deformation as it is pushed into the inlet 220. In this state, the sealing member 221 is unlikely to come loose from the inlet 220.
As shown in
That is, as shown in
The inside diameter of the through-hole 227 of the pushed-in sealing member 224 is smaller than the outside diameter of the support column 226 formed in the diluent chamber 216, and the material of the sealing member 224 is harder than the material of the support column 226. Thus, the support column 226 undergoes elastic deformation, making the sealing member 224 less likely to come loose from the support column 226.
The sealing member 224 is formed from a harder material than the cover 231 and the support column 226. Accordingly, as shown in
Therefore, the channel 217 between the measurement chambers 218 and the diluent chamber 216 is ensured through the operation of blocking off the inlet 220 with the sealing member 221. Consequently, the mixture of diluent and specimen flows from the diluent chamber 216 into the measurement chambers 218.
The pressure receivers 229 of the sealing member 224 have a substantially circular ring shape that includes cut-outs, around the outer peripheral edge of the biological inlet 230.
That is, as shown in
Consequently, even when the sealing member 224 is pushed by the protrusion 228 of the sealing member 221 toward the inside of the diluent chamber 216, a flow path can be ensured in which the opening 225 side of the channel 217 and the diluent chamber 216 are always communicating. Thus, a mixture of diluent and specimen can flow into the measurement chambers 218.
In this embodiment, the biosensor chip 1 into which a specimen has been introduced is inserted through the opening 3 of the biosensor device, and placed on the rotary tray 7. The operator then starts the detection process by giving an input to start up the device from the display component 5 of the device main body 2, which causes the biosensor chip 1 to rotate and exerts centrifugal force on the mixture. Consequently, the centrifugal force causes a specific amount of mixture to flow into the measurement chambers 218. The specimen is then sensed by the optical sensor 12, and the result is displayed on the display component 5.
As discussed above, with the biosensor chip 1 in this embodiment, in a state in which the inlet 220 has been sealed off by the sealing member 221, the sealing component 223 that seals the channel 217 moves downward (in the drawing) in the axial direction of the support column 226, and this opens up the channel 217.
Consequently, even if the biosensor chip 1 is accidentally dropped by the user, since the channel 217 is physically sealed by the sealing component 223, no diluent will flow into the measurement chambers 218. As a result, mixing of the diluent and the specimen just prior to use is prevented, and more accurate detection is possible.
The biosensor device pertaining to the present invention effectively prevents contamination and thereby increases measurement accuracy, and also greatly reduces management costs and other such costs.
1 biosensor chip
2 device main body
3 opening
4 lid
5 display component
6 space
7 rotary tray
8 rotation mechanism
9 shaft
10 temperature sensor
11 heater
12 optical sensor
13 computer
14 controller
15 cover
16 base
16
a film
17 main body case
18 inlet
19 sealing member
20 diluent chamber
21 first channel
22 measurement chamber
23 branched chamber
24 second channel
25 individual measurement chamber
26 reagent mounting hole
28 common reagent
29 individual reagent
31 recess
35 adhesive agent
106 chip main body
107 lower cover
108 inlet
109 upper cover
110 accessory cover
111 sealing member
112 holding chamber
113 first channel
114 dispensing chamber
115 diluent chamber
116 second channel
117 measurement chamber
118 overflow chamber
206 space
207 rotary tray
208 rotation mechanism
209 shaft
210 temperature sensor
211 heater
212 optical sensor
213 computer
214 controller
216 diluent chamber
217 channel
218 measurement chamber
219 main body case
220 inlet
221 sealing member
222 seal
223 sealing component
224 sealing member
225 opening
226 support column
227 through-hole
228 protrusion
229 pressure receiver
230 biological inlet
231 cover
Number | Date | Country | Kind |
---|---|---|---|
2012-112167 | May 2012 | JP | national |
2012-112168 | May 2012 | JP | national |
2012-242425 | Nov 2012 | JP | national |
2012-242426 | Nov 2012 | JP | national |
2012-242427 | Nov 2012 | JP | national |
This is a continuation application under 35 U.S.C. 120 and 35 U.S.C. 365 of International Application PCT/JP2013/002989, with an international filing date of May 9, 2013 which claims priority to Japanese Patent Applications No. 2012-112167 and No. 2012-112168 filed on May 16, 2012 and No. 2012-242425, No. 2012-242426 and No. 2012-242427 filed on Nov. 2, 2012. The entire disclosures of PCT/JP2013/002989 and Japanese Patent Applications No. 2012-112167, No. 2012-112168, No. 2012-242425, No. 2012-242426 and No. 2012-242427 are hereby incorporated herein by reference.
Number | Name | Date | Kind |
---|---|---|---|
7435381 | Pugia et al. | Oct 2008 | B2 |
20040241042 | Pugia et al. | Dec 2004 | A1 |
20060245972 | Osone et al. | Nov 2006 | A1 |
20070003433 | Horike et al. | Jan 2007 | A1 |
20090041627 | Pugia et al. | Feb 2009 | A1 |
20090143250 | Lee | Jun 2009 | A1 |
20100009431 | Cho et al. | Jan 2010 | A1 |
20100086990 | Stanley et al. | Apr 2010 | A1 |
20100158757 | Horiike et al. | Jun 2010 | A1 |
Number | Date | Country |
---|---|---|
2329877 | Jun 2011 | EP |
2006-308366 | Nov 2006 | JP |
2007-500850 | Jan 2007 | JP |
2007-24851 | Feb 2007 | JP |
2007-333716 | Dec 2007 | JP |
2008-281500 | Nov 2008 | JP |
4336834 | Sep 2009 | JP |
2010-25854 | Feb 2010 | JP |
2010-519892 | Jun 2010 | JP |
2011-527753 | Nov 2011 | JP |
2008106719 | Sep 2008 | WO |
Entry |
---|
Focke et al., “microstructuring of polymer films for sensitive genotyping by real-time PCR on a centrifugal microfluidic platform, Lab chip, 2010, 10, 2519-2526”. |
Stumpf et al. “Centrifugal microfluidic system for primary amplification and secondary real-time PCR, lab chip, 2010, 10, 3210-3212”. |
Oosterbroek et al., “EP2329877A1, Machine Translation”, translated on Dec. 13, 2016. |
Office Action dated Jun. 7, 2016 in Japanese patent application No. 2012-242425. |
International Search Report dated Aug. 13, 2013 in International (PCT) Application No. PCT/JP2013/002989. |
Number | Date | Country | |
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20150024477 A1 | Jan 2015 | US |
Number | Date | Country | |
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Parent | PCT/JP2013/002989 | May 2013 | US |
Child | 14501848 | US |