JP2014092421A - Specimen inspection chip and biochemical reaction analyzer using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a specimen inspection chip disposed in a biochemical reaction analyzer capable of detecting a detection target from an injected specimen like blood with high measurement accuracy.SOLUTION: The specimen inspection chip includes: an injection chamber 28 formed in the chip main body for holding the specimen injected from an injection port 27 for injecting a specimen; plural measuring chambers 9 which are connected to the injection chamber via a flow path within the chip main body for generating a mixed liquid reagent by making the specimen flowing in from the injection chamber and a reagent held therein react; and plural marker chambers 26 for measuring fluorescence amount in a portion of the main body on a rotation orbit where the measuring chambers are positioned when the main body performs rotational movement.

Description

  The present invention relates to a specimen test chip and a biochemical analyzer using the same, for example, arranged in a biochemical analyzer in a state in which a specimen such as blood is injected, and detects a detection target from the specimen. In particular, the present invention relates to a specimen test chip and a biochemical analyzer using the same.

  Conventionally, as a specimen testing chip used as described above, a main body having an injection port for injecting a sample, a holding chamber provided in the main body and holding a sample injected from the injection port, and the main body The measurement chamber is connected to the holding chamber via a flow path, and generates a mixed liquid reagent by reacting a sample flowing from the holding chamber with a built-in reagent. (For example, patent document 1).

International Publication No. 2008/106719

  The problem in the above-described conventional example is that when the biochemical analysis is performed using the specimen test chip, the accuracy of measurement of the fluorescence amount is lowered.

  Specifically, when performing biochemical analysis using such a specimen test chip, the optical unit in the biochemical analyzer irradiates the mixed liquid reagent in the measurement chamber with light, and the irradiated mixture The amount of fluorescence emitted from the liquid reagent is measured. At this time, the biochemical analyzer is provided with a temperature adjustment unit for adjusting the temperature in the vicinity of the sample test chip in order to react the mixed liquid reagent in order to increase the measurement accuracy.

  However, assuming miniaturization of the biochemical analyzer, the physical distance between the temperature adjustment unit and the optical unit is close, so that the optical unit is affected by the temperature adjustment unit and the temperature becomes unstable. Along with this, the amount of fluorescence detected by the optical unit changes. Due to this change, the accuracy of measurement of the fluorescence amount of the optical unit is lowered. That is, the measurement accuracy for the specimen test is lowered.

  Accordingly, an object of the present invention is to increase the accuracy of measurement when performing biochemical analysis using a specimen test chip.

  In order to achieve this object, a sample test chip according to the present invention includes a main body having an injection port for injecting a sample, a holding chamber provided in the main body and holding a sample injected from the injection port. A measurement chamber that is connected to the holding chamber through a flow path in the main body, and that generates a mixed liquid reagent by reacting a sample flowing from the holding chamber with a built-in reagent. A marker for measuring the amount of fluorescence is provided on the main body portion on the rotation path where the measurement chamber is located when the main body rotates, thereby achieving the intended purpose. .

  As described above, the sample test chip of the present invention includes a main body having an injection port for injecting a sample, a holding chamber for holding the sample injected from the injection port, and a holding chamber for holding the sample injected from the injection port. A measurement chamber that is connected to the holding chamber through a flow path and that reacts with a sample flowing in from the holding chamber and a built-in reagent to generate a mixed liquid reagent, and the main body rotates. When exercising, the main body portion on the rotating orbit where the measurement chamber is located is provided with a marker for measuring the amount of fluorescence. Therefore, when performing biochemical analysis using this specimen test chip, the accuracy of the measurement is measured. Can be high.

That is, when the main body of the sample test chip in the present invention rotates, the optical unit of the biochemical analyzer irradiates the mixed liquid reagent in the measurement chamber of the sample test chip and emits light from the mixed liquid reagent. The amount of fluorescence emitted from the marker can be measured by providing a marker for measuring the amount of fluorescence on the rotating orbit where the measurement chamber is located. The amount of fluorescence emitted from the marker reflects the influence of the temperature adjustment unit more prominently, so the amount of fluorescence emitted from the mixed liquid reagent can be corrected based on the change in the amount of fluorescence emitted from this marker. As a result, the measurement accuracy of the fluorescence amount can be increased even in an environment where the biochemical analyzer is downsized and the optical unit is influenced by the temperature adjusting unit. That is, the measurement accuracy can be increased.

Overview of biochemical analysis system in an embodiment of the present invention The internal block diagram of the biochemical analyzer 1 in embodiment of this invention FIG. 2 is an internal configuration diagram of a sample test chip 2 in an embodiment of the present invention (A) (b) AA sectional view of specimen inspection chip 2 in an embodiment of the invention. Top view of marker chamber 26 and detection area in the portable embodiment of the present invention (A) Explanatory drawing of the fluorescence amount of the marker in the embodiment of the present invention, (b) Relationship diagram between the concentration of the fluorescent polymer diluted solution and the fluorescence amount Flowchart of measurement of biochemical analyzer 1 in the embodiment of the present invention Explanatory drawing of the change of the fluorescence amount in the embodiment of the present invention Explanatory drawing of the relational expression of the fluorescence amount in the embodiment of the present invention

Hereinafter, a biochemical analysis system according to an embodiment of the present invention will be described in detail with reference to the drawings.
(Embodiment 1)
FIG. 1 is an overview diagram of a biochemical analysis system according to an embodiment of the present invention. This biochemical analysis system includes a biochemical analyzer 1 that detects a detection target from a sample and a sample test chip 2 that can enclose the sample. The biochemical analyzer 1 is a device for analyzing biochemistry in a hospital, a clinic, or the like.

  More specifically, after a sample such as blood is spotted on the sample test chip 2, the sample test chip 2 is set from the chip insertion port 3 provided in the biochemical analyzer 1, and the detection target is detected from the sample. And perform biochemical analysis. A touch panel 4 is provided on the front surface of the biochemical analyzer 1. The user performs a measurement start operation and a measurement stop operation using the touch panel 4.

  FIG. 2 is an internal configuration diagram of the biochemical analyzer 1 in a state where the sample test chip 2 is inserted.

  The biochemical analyzer 1 mainly includes a rotation mechanism unit 5, a temperature adjustment unit 6, an optical unit 7, and a data correction unit 8.

  FIG. 3 is an internal configuration diagram (planar schematic diagram) of the sample test chip 2. The sample test chip 2 includes a plurality of measurement chambers 9 for generating a mixed liquid reagent by holding a sealed sample and reacting with a reagent previously held inside.

  Below, each part which comprises the biochemical analyzer 1 and the sample test chip | tip 2 is demonstrated in detail using FIG.2 and FIG.3 similarly to the above-mentioned.

  The rotation mechanism unit 5 includes a chip holding unit 10 that holds the inserted sample test chip 2 and a motor 11 that is connected to the chip holding unit 10 by a shaft and rotates the sample test chip 2. .

  The temperature adjustment unit 6 includes a housing 12 that covers the sample test chip 2, a heater 13 that heats the inside of the housing 12, and a temperature sensor 14 that measures the temperature inside the housing 12. The temperature adjustment unit 6 can keep the internal temperature of the housing 12 and the temperature of the sample test chip 2 constant by controlling energization of the heater 13 located inside the housing 12.

  The optical unit 7 includes a light source 15, a condenser lens 16, an illumination filter 17, a dichroic mirror 18, an objective lens 19, a fluorescence filter 20, and a detection unit 21. Light from the light source 15 (wavelength 470 nm LED) is collected by the condenser lens 16 and passes through the illumination filter 17. At this time, the illumination filter 17 is a band-pass filter that transmits 430 to 490 nm. Due to its transmission characteristics, although the light with the illumination wavelength of the fluorescent label transmits, the light with the fluorescence wavelength is cut. The passed light is reflected by the dichroic mirror 18, collected by the objective lens 19, and illuminates the mixed reagent stored in the measurement chamber 9 inside the sample test chip 2. The fluorescence emitted from the mixed reagent is collected by the objective lens 19. The condensed fluorescence passes through the dichroic mirror 18 and the fluorescence filter 20. At this time, since the dichroic mirror 18 reflects 430 to 490 nm and transmits 510 to 560 nm, the illumination wavelength is reflected and the fluorescence wavelength is transmitted. The fluorescent filter 20 is a band-pass filter that transmits 510 to 560 nm, and due to its transmission characteristics, the illumination wavelength is cut and the fluorescent wavelength is transmitted. Then, the fluorescence amount of the fluorescence transmitted through the dichroic mirror 18 and the fluorescence filter 20 is measured in the detection unit 21.

  The data correction unit 8 includes an amplifier circuit 22, an analog data processing unit 23, and a digital data processing unit 24. The amplifier circuit 22 amplifies the signal output from the detector 21. The analog data processing unit 23 converts the signal amplified in the amplifier circuit 22 into a digital value. The digital data processing unit 24 performs processing such as collection and analysis of the digital value converted by the analog data processing unit 23.

  Next, the internal configuration of the sample test chip 2 will be described with reference to FIG. The main body of the specimen test chip 2 is made of ABS resin, and as can be understood from FIG. 1, has a substantially rectangular shape with two curved sides, as shown in FIG. A through hole 25 is provided. As shown in FIG. 2, when the sample test chip 2 is inserted into the biochemical analyzer 1, the chip holder 10 is inserted into the through hole 25, and the sample test chip 2 is inside the biochemical analyzer 1. Fixed at. A groove and a reservoir are formed on one side surface of the sample test chip 2 (not shown).

Returning to FIG. 3 and continuing the description, the measurement chamber 9 (ch1 to ch6) and the marker chamber 2
6 (Mark 1 to Mark 4) are arranged concentrically. The measurement chamber 9 and the marker chamber 26 are arranged at equal intervals. The marker chamber 26 is provided with a marker at the bottom, and is used by the optical unit 7 to measure the amount of fluorescence. At this time, the measurement chamber 9 is 3 mm × 4 mm × (depth) 2 mm, and the marker chamber 26 is 1 mm × 4 mm × (depth) 1 mm.

  The flow of the sample will be described using the sample test chip 2. First, the sample is spotted from the injection port 27, and the spotted sample is temporarily stored in the injection chamber 28. The measurement chamber 9 is connected to the injection chamber 28 via a plurality of flow paths, and the sample testing chip 2 is rotated using the rotation mechanism unit 5, so that the sample uses the centrifugal force that is caused by the rotation. And sent to the measurement chamber 9. Here, a reagent necessary for the reaction is sealed in the measurement chamber 9 in advance, and a mixed liquid reagent is generated by mixing the specimen and the reagent using centrifugal force. Further, a film 29 is formed on the upper surfaces of the measurement chamber 9 and the marker chamber 26 as shown in FIG. 4A, and a chamber region is formed inside. The material of the film 29 is acrylic resin (PMMA), the thickness is 0.3 mm, and it is welded using a laser.

  Next, a specific configuration and formation method of the marker chamber 26 will be described.

  First, as shown in FIG. 4A, which is a cross-sectional view of the arc AA of the specimen test chip 2 in FIG. 3, the depth of the marker chamber 26 is a half of the depth of the measurement chamber 9. Thus, the marker chamber 26 is formed. When formed in this way, the optical imaging position coincides with the substantially central portion of the measurement chamber 9 when the optical unit 7 detects the fluorescence amount of these chambers. Since the measurement chamber 9 is present inside so as to be filled with the mixed liquid reagent, the optical imaging position is a measurement target, and can be detected so as to be located near the center of the mixed liquid reagent. As a result, the amount of fluorescence increases and efficient detection becomes possible.

  A specific example of a method for forming the marker chamber 26 for the sample test chip 2 will be described below. First, as shown in FIG. 4A, the specimen inspection chip 2 is formed in a shape in which a portion corresponding to the marker chamber 26 is extracted, and then the fluorescent polymer particle diluent is spotted on the marker chamber 26.

  At this time, the marker chamber 26 is filled with the fluorescent polymer particle diluent. Next, the fluorescent polymer particle diluted solution is naturally dried at room temperature. It is desirable that the humidity be as low as about 40% or less. In this way, the dried fluorescent polymer particles are attached to the side and bottom of the marker chamber 26, and a sufficient amount of fluorescence can be secured. Then, the film 29 is formed by laser welding. After the film 29 is formed, the fluorescence amount of the dried marker is checked. This check is performed by measuring the amount of fluorescence using the optical unit 7 of the biochemical analyzer 1, and confirms whether the amount of fluorescence is within the specification. Finally, the measured fluorescence amount information is affixed to the sample test chip 2 as a barcode.

  Now, in FIG. 4A, the means for realizing the configuration shown by extracting the part corresponding to the measurement chamber 9 and the marker chamber 26 from the sample test chip 2 has been described. Thus, the configuration shown in FIG. 4B is also conceivable.

That is, for the specimen test chip 2, first, through holes are provided in portions corresponding to the measurement chamber 9 and the marker chamber 26. Then, a film 29 is attached to the upper surface of the specimen inspection chip 2 having a through hole by using laser welding. Next, a reagent for reacting the specimen is spotted on the upper surface of the measurement chamber unit 30 as shown in FIG. 5, and then the reagent is dried and fixed on the upper surface of the measurement chamber unit 30. Further, fluorescent polymer particles are applied to the upper surface of the marker chamber unit 31. Then, the measurement chamber unit 30 and the marker chamber unit 31 are attached from the back surface of the sample test chip 2 using an adhesive.

  Even when the specimen test chip 2 is formed in this way, the depth of the marker chamber 26 is one half of the depth of the measurement chamber 9 as in the case of forming as shown in FIG. It forms so that it may become.

  A specific example of the method of forming the marker chamber 26 for the specimen test chip 2 in this case will be described below. First, after the specimen test chip 2 is formed in a shape with a through hole as shown in FIG. 4B, fluorescent polymer particles are applied to the upper surface of the marker chamber unit 31. The fluorescent polymer particles are applied by coating. Specifically, a fluorescent polymer particle dilution liquid in which the fluorescent polymer particles are suspended in a colloidal form is generated, and the generated dilution liquid is applied using an air spray device dedicated to application. As other methods, a static electricity application spray, a dipping method, or the like may be used. After the fluorescent polymer particle diluted solution is attached to the upper surface of the marker chamber unit 31 by any method, it is dried and fixed.

  The fluorescence amount of the marker can be adjusted by changing the concentration of the fluorescent polymer particle diluent or repeating the steps of applying and drying the fluorescent polymer particle diluent.

  Next, the shape when viewed from above the marker chamber 26 is a long hole shape as shown in FIG. Here, regarding the dimension of the marker chamber 26, the length in the circumferential direction of the specimen test chip 2 that performs the rotational motion is the same length as the diameter of the detection area, and the specimen test chip that performs the rotational motion in the same manner. The length in the radial direction in 2 is longer than the diameter of the detection area. In this way, the circumferential direction becomes the operation direction of the marker chamber 26 with the rotational movement of the sample test chip 2, so that the length of the marker chamber 26 in the circumferential direction (short axis) is made equal to the detection area diameter. By setting, disturbance light can be prevented, and the optical unit 7 can detect with higher accuracy. In addition, by setting the length of the marker chamber 26 in the radial direction (long axis) to be longer than the length of the detection area, even when an error occurs when inserting the specimen test chip 2, the fluorescence amount using the marker It becomes possible to reliably perform the measurement. Actually, for example, when the diameter of the detection area is 1 mm, the length of the marker chamber 26 in the radial direction (major axis) may be set to 4 mm and the length in the circumferential direction (minor axis) may be set to 1 mm. .

  Furthermore, regarding the depth of the bottom surface of the marker chamber 26, although the bottom surface is formed horizontally in FIG. 4, the center portion of the bottom surface may be formed deeper than the end portion.

  In particular, this method is used when the area of the bottom surface of the marker chamber 26 is large, or when it is difficult to spot the marker at a position necessary for measurement because the amount of the marker to be attached is small. Thus, the marker to be spotted can be easily concentrated on the center portion, and thus the effectiveness is high.

  The amount of fluorescence of the marker applied to the marker chamber 26 will be described with reference to FIG.

In the present embodiment, the marker in the marker chamber 26 uses four types of markers (Mark1 to Mark4) arranged concentrically. As shown in FIG. 6A, for example, the fluorescence of four markers is used. The amount of fluorescence of these markers is adjusted so that the amount differs with a directly proportional characteristic. This adjustment is adjusted according to the concentration of the fluorescent polymer dilution liquid spotted on the marker chamber 26. More specifically, as shown in FIG. 6 (b), the concentration of the fluorescent polymer diluent in a predetermined amount and the amount of fluorescence of the marker are directly proportional to each other. On the other hand, fluorescent polymer diluents having different concentrations are spotted.

  Below, the flow of the measurement in the biochemical analyzer 1 of this Embodiment is demonstrated using FIG.

  First, a sample such as blood is spotted on the sample test chip 2, the sample test chip 2 is set from the chip insertion port 3 of the biochemical analyzer 1, and a measurement start operation is performed by the touch panel 4 on the front surface of the biochemical analyzer 1. I do.

  Next, an insertion check of the sample test chip 2 is performed (S1). At that time, since a barcode is provided on the curved side surface of the specimen test chip 2 having a thickness of 6 mm, the operator reads this barcode and confirms that the processing chip is inserted. In addition, the type of dry reagent applied to the sample test chip 2 can be read by the barcode. Furthermore, a value indicating the amount of fluorescence before shipment of the marker can be read with the barcode.

  Then, temperature adjustment is started (S2). The interior of the housing 12 that covers the sample test chip 2 is heated by the heater 13, and the heater 13 inside the housing 12 is controlled by the temperature sensor 14. Thereby, the internal temperature of the housing | casing 12 is kept constant (60 degreeC).

  Next, the specimen is filled (S3). The sample testing chip 2 is rotated by the rotation mechanism unit 5 (the number of rotations is 2500 rpm), and the sample is moved to the measurement chamber 9 by centrifugal force.

  Then, the reagent that has been dry spotted in the measurement chamber 9 and the filled sample are stirred by the centrifugal force generated by rotating the sample test chip 2 to generate a mixed liquid reagent (S4).

  Further, the sample inspection chip 2 is rotated by the rotation mechanism unit 5 (rotation speed 240 rpm), and the optical unit 7 is mixed with the mixed liquid reagent filled in the measurement chamber 9 (ch 1 to 6) and the marker ( The amount of fluorescence of Mark 1 to 4) is detected (scanned) (S5).

  Further, the data correction unit 8 calculates the correction coefficients α and β based on the fluorescence amount of the markers (Marks 1 to 4) (S6), and stores the correction coefficients α and β in the measurement chambers (ch1 to 6) based on the correction coefficient. The fluorescence amount of the filled mixed liquid reagent is corrected (S7). The optical unit 7 is detected every minute, and the total time is 40 minutes or more (S8).

  When the fluorescence amount of the mixed liquid reagent filled in the measurement chamber 9 (ch1 to 6) exceeds a certain threshold value, “positive” is displayed on the touch panel 4 as a positive determination. As a determination, “negative” is displayed on the touch panel 4 to inform the user (S9). Finally, the inside of the housing 12 covering the sample test chip 2 is air-cooled by a cooling fan (S10).

  Here, the specific means for correcting the fluorescence amount using the marker in the data correction unit 8 will be described with reference to FIGS. 8 and 9.

As shown in the change diagram of the fluorescence amount in FIG. 8, the optical unit 7 has the fluorescence amount with respect to the mixed liquid reagent in the measurement chamber 9 (ch1 to 6) and the marker in the marker chamber 26 (Mark1 to Mark4). Measure. S1 + 1 to S6 + n and Sm1 + 1 to Sm4 + n are fluorescence amounts measured in the measurement chamber 9 and the marker chamber 26 at times t1 to tn, respectively. The fluorescence amount of the mixed liquid reagent in the measurement chamber 9 (ch1 to 6) and the marker in the marker chamber 26 (Mark1 to Mark4) decreases with time.

  Among these, as shown in FIG. 9, the change in the fluorescence amount for each marker in the marker chamber 26 (Marks 1 to 4) is linear with different slopes before shipment (t = t0) and at the time of measurement (t = tn). It has the properties of Using this property, the amount of fluorescence in the mixed liquid reagent in the measurement chamber 9 (ch 1 to 6) is corrected.

  Specifically, as shown in FIG. 9, a linear relational expression (S = αt0 × M + βt0) is generated based on the fluorescence amounts Sm1 to Sm4 of the marker chambers 26 (Marks 1 to 4) before shipment, and the constants. αt0 and βt0 are calculated. Among these, S is the amount of fluorescence (Sm1 to Sm4), and M is the marker (1 to 4). At the time of measurement (t = tn), a primary relational expression (S = αtn × M + βtn) is generated based on the fluorescence amount (Sm1 + tn to Sm4 + tn) of the marker chamber 26 (Marks 1 to 4). Constants αtn and βtn are calculated, and a correction equation (SM = (αt0 / αtn) × (Sch−βt0) + βtn) of the fluorescence amount of the measurement chamber is derived from these correction equations. Among these, Sch is the measured fluorescence amount (ch = 1 to 6) at the time of measurement (t = tn) of the measurement chamber 9 (ch1 to 6), and SM is the corrected fluorescence amount.

  It should be noted that the fluorescence amount information given before shipment does not need to be given the fluorescence amount information of all the markers, and only the constants αt0 and βt0 described above may be given.

  As described above, the biochemical analysis system according to the present invention can increase the accuracy of measurement of the amount of fluorescence. For example, when detecting a detection target such as a virus from a sample such as blood. It is useful in the biochemical analysis system used.

DESCRIPTION OF SYMBOLS 1 Biochemical analyzer 2 Sample test chip 3 Chip inlet 4 Touch panel 5 Rotation mechanism part 6 Temperature adjustment part 7 Optical part 8 Data correction part 9 Measurement chamber 10 Chip holding part 11 Motor 12 Case 13 Heater 14 Temperature sensor 15 Light source 16 Condensing lens 17 Illumination filter 18 Dichroic mirror 19 Objective lens 20 Fluorescence filter 21 Detector 22 Amplifier circuit 23 Analog data processing unit 24 Digital data processing unit 25 Through hole 26 Marker chamber 27 Injection port 28 Injection chamber 29 Film 30 Measurement chamber Unit 31 Marker chamber unit

Claims (12)

A body having an inlet for injecting a specimen;
A holding chamber that is provided in the main body and holds a specimen injected from the inlet;
In the main body, a sample test having a measurement chamber that is connected to the holding chamber through a flow path and generates a mixed liquid reagent by reacting a sample flowing from the holding chamber with a built-in reagent. A chip,
A sample test chip comprising a marker for measuring the amount of fluorescence in a main body portion on a rotational orbit where the measurement chamber is located when the main body rotates. The specimen test chip according to claim 1, wherein a plurality of the measurement chambers and a plurality of the markers are sequentially arranged in the main body portion. The specimen test chip according to claim 1, wherein the marker is formed at a bottom portion of the marker chamber. The specimen test chip according to claim 3, wherein a film is formed across the upper portion of the measurement chamber and the upper portion of the marker. The specimen test chip according to claim 3 or 4, wherein a depth of the marker chamber is a half depth of the measurement chamber. The specimen test chip according to any one of claims 3 to 5, wherein the bottom portion of the marker chamber is formed so that a central portion thereof is deeper than an end portion. The specimen test chip according to any one of claims 3 to 6, wherein the marker is formed across a bottom portion and a side portion of the marker chamber. 8. The marker chamber according to claim 3, wherein a radial length of the tip in the rotational motion when viewed from above is longer than a circumferential length of the tip in the rotational motion. Specimen test chip. The specimen test chip according to any one of claims 1 to 8, wherein a plurality of the markers are formed, and each marker has a different fluorescence intensity. A biochemical analyzer for detecting a detection target from a sample sealed in the sample test chip according to any one of claims 1 to 9,
A housing for storing the sample test chip;
A temperature adjustment unit for adjusting the internal temperature of the housing;
An optical unit for irradiating light to the measurement chamber for generating the mixed liquid reagent and measuring the amount of fluorescence emitted from the mixed liquid reagent;
A biochemical analyzer having 11. The biochemical analyzer according to claim 10, further comprising a tip rotation holding portion that holds the sample test chip through a through hole provided in the sample test chip when the sample test chip is inserted into the housing. . The biochemical analyzer according to claim 10 or 11, further comprising a data correction unit that corrects the fluorescence amount of the marker measured through the optical unit based on data attached to the specimen test chip.