JP2009204620A - Method for improving throughput in clinical analyzer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an apparatus and a method for detecting patient sample properties and/or analytes in a tip used to aspirate a patient sample liquid and to dispense it onto a slide test element. <P>SOLUTION: Spectrophotometric analysis is performed on the liquid, while still in the tip, by scanning the tip for transmittance in a light-tight enclosure using near infrared and adjacent visible radiation to detect the absorbance spectra of the liquid. Thereafter, or prior thereto, the liquid is dispensed onto a dried slide test element for assaying the analytes that are not assayed spectrophotometrically, thus throughput is improved. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a new use of an old device and a new dispensing station that allows spectrophotometric analysis to be performed on a blood sample prior to examining the blood sample in a conventional dry or wet assay.

In general, spectrophotometric analysis is applied to many liquids to determine the contents. Such analysis is particularly effective when performed with near-infrared radiation due to the ability to differentiate between near-infrared radiation target analytes and other substances.
It is clear from Non-Patent Document 1, for example, that spectrophotometric analysis can identify hemoglobin, albumin, lipids, and many other serum components.

European Patent Application Publication No. 185330 US Pat. No. 3,992,158 US Pat. No. 5,441,895 Canadian Patent No. 2011511 U.S. Pat. No. 4,340,390

Clinical Chemistry (Clin. Chem.) 38, 1623-1631 (1992) Anal. Chem. Vol. 59, No. 17, 1007A-1017A (September 1987) Analytical Chemistry Vol. 66, No. 15, 795A-804A (August 1994)

  However, there are problems in applying such analysis to blood samples to determine the content or characteristics of the blood sample. For example, it has been difficult to apply this analysis to a blood sample because the blood sample is initially taken, ie, in the primary patient collection container. These containers are usually tubes of various sizes and have been centrifuged to separate liquid serum or plasma from the cell phase. Thus, such tubes require a) patient identification labels, b) various unpredictable locations of serum, and c) large volumes (milliliters) of samples. For various locations, there is no doubt that an incorrect evaluation will occur if the gel barrier used to separate the liquid phase from the cell phase is scanned instead of the liquid phase.

  Therefore, when handling a tube of liquid with an unpredictable height, it is necessary to make an extra exposure and take extra time to divide it into secondary tubes, or for example, as shown in FIG. It has been practical to determine where the liquid phase is by LED scanning the contents of the tube. Such requirements cause additional equipment costs and processing delays. This, combined with the difficulty of scanning spectrophotometrically through the patient label, makes such scanning of the primary collection container problematic and expensive.

  On the other hand, conventional clinical analyzers that test target substances using dry slide test elements typically require at least 5 minutes to perform target substance assays if incubation is required. And With these incubation times, it becomes difficult to obtain a throughput well over 1000 tests per hour. There is a strong need for methods that allow higher throughput in such analyzers.

  Thus, prior to the present invention, an inexpensive and simple method for spectrophotometric scanning of biological fluids such as serum or plasma separated from whole blood, i.e., what container is being used. There has been a need to provide a method that eliminates the need to find the location of the liquid without, and further eliminates the need to scan through the identification label. Furthermore, there is a need to improve the inspection throughput in the analyzer for assaying the target substance.

According to the invention, in a clinical analyzer comprising a dispensing station and at least one examination station for detecting a target substance in a patient sample, the dispensing station comprises:
An aspirator probe;
A tip attached to the probe for collecting biological sample liquid from a primary collection container and dispensing at least a portion of the collected sample liquid onto or into a test element;
Means for generating a partial pressure or a partial vacuum in the probe and the tip,
a) aspirating biological fluid into one of the tips attached to the probe;
b) The biological fluid is in the tip and the tip is attached to the probe by passing the tip through the tip at a plurality of wavelengths with light of near infrared and near visible radiation wavelengths. Detecting one or more target substances in the solution and correlating the transmitted light with the concentration of the one or more target substances in the biological fluid, and Performing a spectrophotometric analysis at a wavelength, the correlating step comprising calculating a value of a first derivative of absorbance as a function of wavelength,
c) dispensing a portion of the biological fluid from the tip into the test element;
d) inspecting a test element to which a biological fluid has been added at the test station for a target substance other than the one or more target substances, wherein the one or more target substances are A method is provided for improving throughput in a clinical analyzer that increases throughput by the amount of time that is no longer needed for testing at a testing station.

FIG. 3 is a partial isometric view of the aspirator probe of the analyzer showing the position of the scan lock of FIGS. FIG. 3 is an alternative partial elevation view in an intermediate cross section of the device showing two alternative embodiments of the present invention. Partially intermediate cross section showing air holes that prevent both pulsating upward pressure when the tip is inserted and suction at the end of the tip when the tip is removed FIG. 3 is an isometric view of the inspection station 82 of FIG. FIG. 6 is a partial isometric view of another alternative embodiment of the present invention. FIG. 5 is a cross-sectional plan view of a portion of the structure of FIG. 4 showing a mechanism for determining the position of a chip 48A. 4 is a regression plot of hemoglobin levels of a test sample scanned and analyzed spectrophotometrically according to the present invention. 4 is a regression plot of hemoglobin levels of a test sample scanned and analyzed spectrophotometrically according to the present invention. 4 is a regression plot for the level of lipemia in a test sample scanned and analyzed spectrophotometrically according to the present invention. 4 is a regression plot for the level of lipemia in a test sample scanned and analyzed spectrophotometrically according to the present invention. 4 is a regression plot for the level of jaundice of a test sample scanned and analyzed spectrophotometrically according to the present invention. 4 is a regression plot for the level of jaundice of a test sample scanned and analyzed spectrophotometrically according to the present invention. 2 is a plot of spectral transmission detected by the present invention showing samples of jaundice, hemoglobin, and lipid, respectively. FIG. 3 is a partial elevation view of an alternative embodiment of a light-tight enclosure similar to FIG. 2 (A). FIG. 3 is a partial elevation view of an alternative embodiment of a light-tight enclosure similar to FIG. 2 (A). FIG. 4 is a regression plot for the level of bilirubin in a test sample scanned and analyzed spectrophotometrically according to the present invention. FIG. 4 is a regression plot for the level of bilirubin in a test sample scanned and analyzed spectrophotometrically according to the present invention. FIG. 17 is a regression plot for the analysis of hemoglobin, similar to FIGS. 15 and 16. FIG. 17 is a regression plot for the analysis of hemoglobin, similar to FIGS. 15 and 16.

  The present invention is described below with reference to preferred embodiments, in which a suitable (conventional) translucent disposable tip performs analysis on a target that represents patient sample quality of serum or plasma. It is used with a suitable (conventional) analyzer aspirator and a suitable light impervious enclosure connected to the spectrophotometer by a passage using an optical fiber. In addition, however, the present invention provides a translucent or transparent tip, aspirator, liquid, or light-tight enclosure type, spectrophotometer used as long as the target has sufficient NIR and near-visible radiation absorption. It is used regardless of the optical path that is the light path and the optical path from which it is, and the target substance to be detected. That is, the target substance may be a conventional substance that is tested for concentration on a slide test element in a conventional analyzer, such as albumin or glucose. The fluid may be whole blood, urine, or cerebral spinal fluid. The liquid is preferably not limited to reagents. In addition, the tip may be non-disposable and may be used continuously, and an open lens system may be used in place of the optical fiber to collect the light to the light-tight enclosure and then to the detection station.

  The spectrophotometer used with the present invention is not shown in the following and is not described in detail. The reason is that any spectrophotometer works as long as the spectrophotometer generates radiation in the near-infrared and near-visible light regions with sufficient spectral accuracy and detects it through transmission. Because. As used herein, “near infrared and near visible” means radiation between 400 nm and 2500 nm, most preferably between 475 nm and 1075 nm. These wavelengths are advantageous because they exhibit sufficient spectral transmission of the disposable tip as well as sufficient spectral absorption from the target analyte. 475 nm is believed to be particularly effective for bilirubin detection according to the present invention. Effective tip materials that allow the desired spectral transmission are those commonly used to make disposable tips (polypropylene or polyethylene).

  Further, as used herein, “spectrophotometry” (or spectrophotometry) refers to a method of capturing a spectral response for a range of wavelengths and correlating the response for each wavelength in that range. By comparison, “photometry” means the analysis of light radiation that correlates the response to only a specific wavelength. Therefore, the “spectrophotometer” is an apparatus that performs this spectrophotometric analysis.

  Also, as used herein, a “primary patient collection container” is a patient's biological fluid, usually blood, that is first labeled and placed in order to prepare the desired sample solution for testing. Means a container to be treated. In the case of whole blood, such treatment involves phase separation in which liquid serum or plasma is separated from the cell phase, which usually consists of blood cells at a gel separation barrier.

  Furthermore, the “test element” used in the present application means an arbitrary container in which at least one reagent is supplied in advance, and this container is, for example, a so-called dry slide test element as described in Patent Document 2, Alternatively, such as a cup or well having a well pre-coated with one or more antibodies or an uncoated well to which a reagent has been added as described in US Pat.

Also, “impervious to light” as used in this application means that it has the effect of removing ambient light to such an amount that only 10% of the detected light is due to external ambient light.
Furthermore, as used herein, “having jaundice” means a state where high levels of bilirubin and / or biliverdin are present in the sample.

  Details regarding the mathematical analysis required to correlate the transmission of near-infrared and near-visible radiation biological fluids with the concentration of the target substance are not described. The reason for this is that such mathematical analysis is publicly known as apparent from the editorials of Patent Literature 4, Non-Patent Literature 1, and the teaching literature of Non-Patent Literature 2 and Non-Patent Literature 3. is there.

  FIG. 1 shows a conventional analyzer 12 utilizing the latest invention. Conventionally, the dispensing station 18 is used to feed a biological fluid, such as serum or plasma, from a feed consisting of a plurality of primary collection containers 19 on a tray 20 to a disposable tip 48 attached to an aspirator probe 46. Samples are collected by aspiration. Subsequently, the sample liquid is obtained from a slide test element supply source (not shown) and dispensed into the slide test element E held in the slide distributor 30. The dispenser 40 including the probe 46 is controlled by a mechanism such as a vertical driving device 44 and a movable base 42 attached to the support rod 70, and all such mechanisms are described in, for example, Patent Document 5. Yes. Any kind of conventional pump 71 can be used as a means for creating a partial vacuum or partial pressure in the chip 48.

  According to one aspect of the present invention, the tip 48 is used in a novel way other than simply collecting the liquid from the container 19 by suction and subsequently dispensing it into the slide test element E. The tip 48 carrying the aspirated sample liquid is moved to the inspection station 82 in the direction of arrow 80 in FIG. 1 before being placed on the holder 117 for dispensing. As more clearly shown in FIGS. 2 (A), 2 (B) and FIG. 3, the inspection station 82 is an enclosure that has a light-impervious effect having a cavity 84 that is dimensioned to receive the chip 48. A scanning block is provided. Preferably, the cavity 84 includes an upper portion 86 in FIG. 3, a lower portion 88 having a smaller inner diameter than the upper portion 86, a ledge 90 at the boundary between the upper and lower portions, and an air hole 92 in the ledge 90. And a conical exit hole 94 extending away from the lower portion in a direction away from the upper portion and two passages 96 suitable for receiving optical fibers 98, 98 'towards and coming from the spectrophotometer portion. . The outlet hole 94 is formed in a shape that substantially matches the shape of the outlet hole portion of the tip 48, and thus in the case of this preferred tip 48. An optional air tube 100 is connected to the outlet hole 94 to reduce the possibility of pumping fluid out of the tip. If the air tube is opaque, optionally, the air tube helps eliminate light leakage to the chip.

As shown in FIG. 1, optical fibers 98, 98 ′ are connected to a conventional spectrophotometer comprising a light source 110 and a detector combined into a single device 110.
When maximally effective, the inspection station 82 has the effect of impinging light as defined herein, and the light passing through the detector passes through the optical fiber 98 through the chip 48. At least 90%. There are several ways in which this can be achieved.

  First, as shown in FIG. 2A, an upper surface 90 that acts as a support shoulder for the enlarged upper portion 111 and thus does not go higher than the enlarged upper portion 111. In the case of an inspection station 82 having a block 83 with a side gap of 0.5 mm between the chip 48 and the chip 48, the leakage of light that occurs is transmitted by the optical fiber 98 with NIR and near-visible radiation. Is corrected by taking an empty reading under the same ambient light conditions that are used when the fiber 98 is transmitting no light. The empty reading is then subtracted from the sample reading and the reference reading.

Alternatively, if the subtracted empty reading should not be utilized and the side gap is still the same as described above, the same light opacity can be blocked as shown in FIG. This can be achieved by extending the height of 83 to at least the height of the upper surface 113 of the upper portion 111 of the chip 48.
Since the seating of the tip 48 on the shoulder surface 90 provides an effective seal, preferably some sort of air venting is made between the upper portion 86 and the lower portion 88 when the tip 48 is inserted and withdrawn. This is the function of the air hole 92 in FIG. This air hole relieves the increase in pressure that occurs when the tip is inserted into the inspection station 82 so that bubbles cannot be forced through the tip's liquid and interfere with optical scanning of the liquid. . Similarly, after the light is scanned, the air holes 92 prevent a vacuum from being generated when the tip 48 is withdrawn, but this vacuum may draw a portion of the sample liquid from the tip 48, Contamination of inspection station 82 for the next chip and sample.

In order to further assist in centering the tip 48 in the cavity 88 between the optical fibers 98 and 98 ', as shown in FIG. A plurality of positioning protrusions 140 can be arranged.
In use, the tip 48 is inserted into the inspection station 82 prior to insertion into the holder 116. While in the inspection station 82, the NIR and near visible wavelength rays defined above are passed through the tip and liquid so that the transmitted radiation is spectrophotometrically analyzed by the spectrophotometer 110. The signal produced by the detector is then calculated for correlation with the concentration of the target substance. Suitable sets of target substances are those that measure sample properties, in particular those selected from the group consisting of hemoglobin, lipids, bilirubin (BR), and biliverdin (BV) as shown in the examples below. is there. However, any target substance that can be spectrophotometrically detected by the absorption spectrum can be detected with the correlation calculated by the present invention. More specifically, certain assays that have been conventionally performed on the slide test element E can be performed spectrophotometrically through the chip as described below.

Thereafter, the chip is withdrawn and inserted into the holder 116, at which point the sample solution is conventionally loaded with one or more reagents to confirm the concentration of the analyte in the sample solution as is known. Dispensing into the containing slide test element E.
As will be readily apparent, the chip 48 used in the present invention can transmit NIR and near visible radiation, most preferably radiation from 475 nm to 1200 nm, and labeling is performed only in the primary container 19. Since it is not, there is preferably no label. Effective materials for this purpose include polypropylene and polyethylene.

  The inspection station 82 need not be configured as a solid block having only disposable tip cavities and optical fiber holes, and the tips need not be lowered to the same location. Instead, the side wall of the inspection station 82 is opened and closed, and as shown in FIGS. 4A and 5, a slot that allows the chip to pass through can be provided. Parts that are similar to those previously described are given the same reference numbers, and a suffix “A” is appended to distinguish them from these reference numbers.

  Thus, inspection station 82A comprises two fixed opposing portions 109, 112 that are spaced apart by a distance. Each portion has an opposing surface 116, 116 ′ that defines a slot 115 therebetween. The upper surface 117 of the surfaces 116, 116 'provides a pedestal for the guide rail and upper portion 111A of the tip 48A. Portion 109 has an optical fiber 98A passing through portion 109 from a light source (not shown), while portion 112 has a sensor 114 connected to a spectrophotometer on surface 116, the spectrophotometer being portion 112. Or connected to the portion 112.

The opposing surfaces of portions 109 and 112 define a slit 115 spaced apart by a distance that allows the disposable tip 48A to slide through as indicated by arrow 120. These opposing surfaces can be separated by a certain distance for slipping of the tip 48A.
Portions 109 and 112 create slots with aspect ratios that are significantly smaller than the aspect ratio described above with respect to the vertical hole 84 for the passage of the tip 48A indicated by arrow 120, so for spectrophotometric measurements. It is better to close the slot 115. To this end, as indicated by arrows 136 and 138, a portion indicated by reference numeral 134 is a pivoting door 130, 132 wide enough to close the slot 115 when pivoted to a closed position (not shown). And hinged to both ends of the portion 109. (Door 132 is shown in phantom lines for simplicity only). In order to pivot the door, a hinge 134 pintle is preferably attached or attached to a rotary drive shaft (not shown) of a conventional motor 136.

Alternatively, doors 130 and 132 can be omitted by lengthening slot 115 to have a horizontal aspect ratio equal to the vertical aspect ratio described above with respect to cavity 84.
To assist in accurately stopping the lateral movement of tip 48A as indicated by arrow 120 at optical fiber 98A and detector 114 shown in FIG. 5, a spring-loaded detent is preferably provided. 210 is provided on the surface 116 and this detent 210 cooperates with a fixed projection 212 on the opposite surface 116 '. After reading, when moving the tip 48A from the slot 115 in the direction of the arrow 120, the detent 210 is pressed against the surface 116 by the tip. As stated in the previous embodiment, chip 48A transmits the NIR and near visible wavelengths used.

  Alternatively, the portion 112B shown in FIG. 4B can be movably attached to the plate 122B to block light leakage. Parts similar to those previously described are given the same reference numbers, and a suffix “B” is attached to distinguish them from these reference numbers. Accordingly, inspection station 82B includes a plate 122B that forms a U-shaped slot with surfaces 116B and 116'B, which U-shaped slot is shown by arrow 120B at the same time that chip 48B is supported on top surface 117B. Makes it possible to slip through. Optical fiber 98B transmits light through stationary portion 130B, and sensor 114B on stationary surface 116B transmits light to a spectrophotometer not shown.

  To block possible light leakage through the U-shaped slot consisting of plate 122B and faces 116B and 116'B, portion 112B is mounted to slide on plate 122B and is driven by arrow 162 168 by rack 162 and drive pinion 164. Open or close the slot. When closed, face 116B and tip 48B occupy space 172 in portion 112B and wall portion 169 closes slot 115B.

  In addition to examining patient sample characteristics, any target substance that can be spectrophotometrically analyzed using NIR and near visible wavelengths is analyzed by the spectrophotometer 110 while the patient sample is on chip 48A. Can do. These target substances include, among others, hemoglobin, albumin, and glucose. By inspecting these target substances in the chip, but not necessarily, in practice, when the sample is placed on the slide test element E, preferably the analyzer is further further on those target substances. Skip the test. Since the spectrophotometric detection through the chip requires only 4 seconds for all the target substances to be analyzed compared to 4 seconds for each individual assay performed on the slide test element E, this means that Increase throughput significantly. The “time to produce results” is further dramatically improved by spectrophotometric analysis through the chip, which is 4 seconds through the chip compared to 5 minutes with the slide test element.

  As an example of improved throughput, the following is an estimate of the benefits that can be achieved with an analyzer such as that sold by Johnson & Johnson Clinical Diagnostics under the trademark “VITROS950” analyzer. This estimation is based on the fact that dispensing of the sample liquid to the slide test element is a limiting step in the analysis, and the dispensing step is 8 seconds for aspiration, dispensing the test element to the VITROS950 (trademark) analyzer. It is assumed that 4 seconds or the like is required for loading into the distributor, and that the colorimetric analysis is performed in the chip according to the present invention.

  If the mixing ratio of the chemical properties to be performed is a potential difference test 0, a colorimetric test 7, and a differential test 0, the throughput is 300 inspection elements per hour when the present invention is not used. With the present invention, the throughput is shown to be 2100 inspections per hour, a 7-fold increase. On the other hand, if only the colorimetric inspection 5 is performed, either the differential inspection 2 or the potential difference inspection 2, the throughput without using the present invention is 420 inspections per hour, and with the present invention, the throughput is 1050 inspections per hour. , An increase of 2.5 times. Furthermore, if the mixing ratio of the seven chemical properties is performed only in the colorimetric test 3 and in the potential difference test 4, there is no increase in throughput obtained by carrying out the present invention (both in 525 tests per hour in both cases). is there).

  Preferably, such an inspection of such analyte while in the chip is performed with some temperature control of the sample liquid. This not only needs to be done by controlling the temperature at the inspection station 82, but can also be done by heating or cooling the sample liquid in the container 19 of FIG. Can be done by heating or cooling while in the chip 48 or the like, but at the inspection station 82 no heating or cooling is done.

  However, there are still some assays that require the use of slide test element E. The process is shown schematically in FIG. Tip 48 is inserted into holder 117 and a portion of the patient sample is dispensed into slide test element E. Thereafter, the dispenser 30 is rotated in arrow 140 to a position where the test element E is linearly transferred to the incubator (not shown) in arrow 142 and the test element E is read or detected in the test station 146 in the incubator. Until it is rotated to arrow 144, all of which are known and conventional. The inspection station 146 is conventionally provided with a colorimetric or potentiometric detector as opposed to the spectrophotometer 110 used with the chips 48, 48A.

As described above, preferably, the test performed at the test station 146 skips the test performed through the chip, but such a spectrophotometric assay is used to “confirm” the accuracy of the test performed within the chip. It is also possible to repeat at the inspection station 146.
It is also conceivable to reverse the order of inspection, i.e., a portion of the sample solution can be placed on the inspection slide as described above before taking measurements through the chip at NIR and near visible wavelengths.

To illustrate the present invention, the following non-consumable inspection is performed.
The device of FIG. 2 is used, sold by Johnson & Johnson Clinical Dyganotics under the trademark “Vitros”, formerly known as the “Ektachem” disposable chip. A disposable tip was used. The optical fiber is a 0.2 mm single fiber, which connects inspection station 82 to a “TC2000” dual-beam synchronous spectrophotometer via fibers 98 and 98 ′, which is a CME telemetry ( A linear diode array detector using a tungsten-halogen incandescent light source 110 is used as detector 112 sold by Telemetrix). In order to be able to detect only radiation from 580 nm to 1100 nm, a diffraction grating was used in the detector 112. (The reference beam portion of the spectrophotometer is omitted for clarity). The amount of liquid sucked into the chip 48 was 50 μL so that the height of the liquid was completely above the passing arrow 200 in FIG. Only 30 μL of proven need be tested.

  The liquid to be tested is primarily calibrated as hemoglobin, Intralipid (trademark) sold by Pharmacia (fat emulsion that mimics naturally occurring milky fat granules), biliverdin, Random combinations of known amounts of fluids containing all of these fluids added to human serum substrates.

  Table 1 below shows Hb, IL, and BV in the serum after addition. “Hb” means hemoglobin, “IL” means Intralipid ™, “BV” means biliverdin, and “BR” means bilirubin.

  A second combination of 21 liquids similarly prepared was prepared to have the components in Table 2 and treated as unknown.

The first combination of liquids was irradiated as described above to create a calibration algorithm using a conventional spectrophotometric technique, and the Hb value detected in this measurement is shown in FIG. 6 to obtain a regression graph. The actual values were filled in the graph. Various calibration algorithms are available, such as selecting the formula by first and second derivatives. The following equation is merely exemplary.
1) Hb (g / L) = C ₁ (dA ₆₀₀ / dλ ₆₀₀ ) −C ₂ (dA ₆₆₃ / dλ ₆₆₃ ) −C ₃
2) IL (g / L) = C ₄ (dA ₈₇₄ / dλ ₈₇₄ ) + C ₅
3) BV (mg / dL) = C ₆ (dA ₇₂₄ / dλ ₇₂₄ ) −C (dA ₈₀₃ / dλ ₈₀₃ ) + C ₈
Where A ₆₀₀ is the absorbance at ₆₀₀ nm, λ ₆₀₀ is the wavelength of ₆₀₀ nm, and other values of A and λ are the same, and (dA _i / dλ _i ) is the first derivative of absorbance with respect to wavelength, C ₁ , ... C ₉ is preferably a constant having the following values:

C ₁ = 15.892 C ₅ = 0.244
C ₂ = 15.882 C ₆ = 98.068
C ₃ = 0.21 C ₇ = 122.732
C ₄ = 252.155 C ₈ = 0.0685
The regression correlation coefficient R ^{2 in} the case of FIG. 6 was 0.991.

Next, the second combination of liquids is irradiated as described above, and using the calibration algorithm of FIG. 6 derived from the first combination of liquids, the predicted values become known results as in FIG. On the other hand, it was filled in the graph. An R ² value of 0.982 is excellent. This accuracy is suitable to allow this result to be reliably applied to clinical assays for Hb in unknown samples, instead of testing on slide test elements.

Similarly, the spectral values detected as described above were evaluated for IL. The result of calibration is shown in FIG. 8, and the result of prediction is shown in FIG. R ^{2 in} this case is 0.9941 and 0.9878, respectively.
Also, the described spectrum was evaluated, but this time the analysis was for BV. FIG. 10 shows the result of the calibration, and FIG. 11 shows the result of the prediction such that the value of R ² is shown.

  A new, third liquid combination was prepared to illustrate the present invention with respect to bilirubin detection, and the calibration liquid format for that combination was configured as shown in the table below.

The calibration algorithm used for this test was as follows.
4) BR (mg / dL) = C ₉ (dA ₄₉₅ / dλ ₄₉₅ ) + C ₁₀ (dA ₅₁₂ / dλ ₅₁₂ ) + C ₁₁ (dA ₅₇₈ / dλ ₅₇₈ ) −C ₁₂
The constants are as follows:
C ₉ = −24.878
C ₁₀ = 201.61
C ₁₁ = 44.98
C ₁₂ = 6.475
A fourth combination of liquids was similarly prepared for confirmation regarding the prediction of bilirubin values, and the combinations were organized as in the table below.

The spectrum was evaluated as in the previous example. FIG. 15 shows the result of the calibration, and FIG. 16 shows the result of the prediction such that the value of R ² is shown.
For all four experiments (Hb, IL, BV, and BR), the results are excellently correlated, and the results are sufficient to be used in place of testing with the slide test element, any of which is desired Should be regarded as a test of. In any case, these results clearly make it possible to confirm the characteristics of a sample of biological fluid and fail if it is determined that the sample is outside the range of acceptable characteristics. To be able to.

As examples of other calibration algorithms that can be used, the following for IL is an alternative to equation # 2 above:
2 ′) IL (g / L) = C ₁₃ (dA ₉₉₉ / dλ ₉₉₉ ) + C ₁₄ (dA ₁₀₅₁ / dλ ₁₀₅₁ ) −C ₁₅
Here, C ₁₃ = 166.068, C ₁₄ = 92.352, and C ₁₅ = 0.693. If this calibration algorithm is used for the first and second liquid combinations described above, R ² will be 0.988 for calibration and 0.984 for prediction. (The actual plot is not shown).

  Referring again to FIG. 7, it becomes clear that the method for confirming the presence of hemoglobin described above showed linearity, ie, measurement accuracy up to 5 g / L. If it exceeds 5 g / L, the data tends to move away from the “true value”. This is most clearly shown in FIG. Therefore, if an amount greater than 5 g / L is contained, another combination of algorithms is preferred.

  More specifically, if two different algorithms are used to cover the entire range, better followability with the true curve is achieved, and these algorithms produce continuous data at the overlap. Selected as shown. Preferably, the algorithm used from 3 g / L up is the natural logarithm of hemoglobin concentration in g / L units. Most preferably, from 3 g / L to 35 g / L, the algorithm used is of the form:

Here, detailed examples of C ₁₆ , C ₁₇ and C ₁₈ are C ₁₆ = −17.38 and C ₁₇ = + 9.96 + C ₁₈ = + 0.37.
Below 3 g / L, various algorithms of a type similar to Equation 1) shown above are effective. In order to mesh with formula 5) above 3 g / L, a very preferred formula is:
1 ′) Hb (g / L) = C ₁₉ (dA ₅₉₃ / dλ ₅₉₃ ) + C ₂₀ (dA ₆₀₈ / dλ ₆₀₈ ) + C ₂₁
Here, C ₁₉ is preferably -66.689, C ₂₀ is preferably 81.081, and C ₂₁ is preferably -0.25.

  When the hemoglobin levels shown in FIG. 17 were recalculated using the above-described new algorithms 5) and 1 ') above 3 g / L and below 3 g / L, the plot of FIG. 18 resulted. This plot shows that the predicted value is closer to the “true” straight line at 45 ° than that obtained at the data points in FIG.

Although the data in FIG. 18 only extends to 10 g / L, it was shown that using the no dilution step, Equation 5) and Equation 1 ′) are valid up to 35 g / L for hemoglobin values.
FIG. 12 shows that the values of the first derivative of the absorbance in the NIR and near-visible spectra are sufficiently separated to allow independent detection of samples where IL, BV, or Hb components are present at the effective wavelength. It is a plot which shows having done. That is, curve 200 is a sample without any of those components, curve 202 is a sample with only 1.79 g / L Hb, curve 204 is a sample with only 2.38 g / L IL, curve 206 is 3. Sample with only 95 mg / dL BV. Accordingly, Hb mainly contributes to the NIR region of 580 to 605 nm, IL contributes to the region of 896 to 1051 nm, preferably 896 to 939 nm, and BV contributes to the region of 680 to 750 nm.

  In yet another embodiment, the chip is not altered from a conventional chip, but before the absorption spectrum of FIG. 13 is received, more than one NIR and near visible radiation chip is passed by the spectrophotometer. Passing is achieved. The aforementioned parts are referred to by the same reference numerals, and a suffix “D” is attached to distinguish the reference numerals.

  Accordingly, the chip 48D is fitted into the cavity 84D as described above so that the irradiation with the NIR and near-visible radiation emitted from the optical fiber 98D is received by the processing optical fiber 98'D. However, unlike the previous embodiment, the light receiving optical fiber 98'D is not in a position to receive "first pass" radiation unless it directly faces the light emitting optical fiber 98D. Instead, at least one and preferably three pairs of mirrors (230, 232; 240, 242; 250, 252) are arranged to re-pass the tip 48D through the radiation as many times as the mirrors. (Three pairs of six mirrors allow the tip to penetrate the radiation six times).

  In yet another embodiment, the optical fibers 98, 98 '(or other types of optical fibers disclosed above) need not allow NIR and near visible light to pass through only the thickest portion of the chip. Instead, light can be transmitted through a narrower neck. (Similar parts to those previously described bear the same reference numbers and are appended with a distinguishing suffix “E”).

  Accordingly, the irradiating optical fiber 98E of FIG. 14 is placed in the block of the inspection station 82E to irradiate the conical neck portion 300 of the tip 48E, which is compared with the diameter “d” of the body portion 224E. Then the diameter becomes smaller. The light that is transmitted through the tip to the receiving optical fiber 98'E then passes through a much narrower portion of the sample. This is desirable if the analyte to be detected is of high density or has a high extinction coefficient for the NIR and near visible wavelengths. In the most extreme case, the optical fibers 98E and 98'E are moved down to the virtual position 302 for reading through the narrowest portion 304 of the chip 48E.

  Alternatively, the passage of NIR and near-visible radiation through a narrower portion of the tip simply raises the tip (and probe) sufficiently within the inspection station 82 and irradiates with NIR radiation using the previous embodiment. Can also be achieved. More preferably, the order of steps is in a light-tight enclosure including NIR and a near-visible radiation source as shown in any of FIGS. 2 (A) (B), 4 (A) (B). Lowering the chip until the chip is seated; scanning the chip and its contents with NIR and near-visible radiation emitted from the radiation source; if the contents have a density above a predetermined threshold, then the radiation source The sequence is to lift the chip in the enclosure until it is arranged to scan a narrower part of the chip.

It is an advantageous feature of the present invention to improve the throughput of the target substance assay in the analyzer.
Yet another advantageous feature is that the results are obtained in a shorter time since no incubation time is required for the spectrophotometric analysis.

12 Analyzer 18 Dispensing Station 19 Primary Collection Container 46 Aspirator Probe 48 Tip 71 Pump 82 Inspection Station

Claims (3)

A method for improving throughput in a clinical analyzer comprising a dispensing station and at least one examination station for detecting a target substance in a patient sample, comprising:
The dispensing station is
An aspirator probe;
A tip attached to the probe for collecting biological sample liquid from a primary collection container and dispensing at least a portion of the collected sample liquid onto or into a test element;
Means for creating a partial pressure or a partial vacuum in the probe and the tip;
The method comprising:
a) aspirating biological fluid into one of the tips attached to the probe;
b) transmitting the light at near infrared and near visible radiation wavelengths at a plurality of wavelengths while the biological fluid is in the chip and the chip is attached to the probe, Permeate the chip by detecting one or more target substances in the biological fluid and correlating the concentration of the one or more target substances in the biological fluid with the transmitted light. Spectrophotometric analysis of a portion of the light at a plurality of wavelengths, wherein the correlation includes calculating a value of a first derivative of absorbance as a function of wavelength;
c) dispensing a portion of the biological fluid from the tip into a test element prior to performing the spectrophotometric analysis;
d) inspecting the test element with the biological fluid added at the test station for target substances other than the one or more target substances;
Increasing the throughput by the amount of time that is no longer required to inspect the one or more target substances at the inspection station;
The correlating step comes to cover the entire range using two calibration algorithms, one calibration algorithm used from 0 to about 3 g / L and another calibration algorithm used at 3 g / L and above. A method for improving throughput in a clinical analyzer.   The method of improving throughput in a clinical analyzer according to claim 1, wherein the other calibration algorithm obtains the natural logarithm of hemoglobin concentration from the value of the first derivative of absorbance with respect to wavelength. The two calibration algorithms are:
An equation for hemoglobin concentration in the range of 0 to about 3 g / L;
Hb (g / L) = − 66.689 (dA ₅₉₃ / dλ ₅₉₃ ) +81.081 (dA ₆₀₈ / dλ ₆₀₈ ) −0.25
Formula for hemoglobin concentration in the range of 3 g / L or more,
Hb (g / L) = e ^{[-17.38 (dA593 / dλ593) +9.96 (dA608 / dλ608) +0.37]}
A method for improving throughput in a clinical analyzer according to claim 2 comprising:

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