Device for spreading or staining and determining sedimentation rate

Abstract

Device for spreading or staining and determining sedimentation rate A device for spreading or staining and determining sedimentation rate comprises a first group (4) arranged to take a blood sample from a tube and to carry out a smear on this sample, and a second group (6) arranged to take a blood sample from a tube and to carry out a determination of sedimentation rate.The device comprises at least one sampling member (8) controllable for operation by the first group (4) and operation by the second group (6) for sampling a blood sample such that a sample taken for the first group (4) is not used by the second group (6), and a sample taken for the second group (6) is not used by the first group (4), the second group (6) being provided with a sensor (20) comprising an infrared light source (12) and an optical sensor (14) arranged substantially opposite each other around a tube (16) connected to an outlet end of the at least one sampling member (8) such that the light emitted by the infrared light source (12) reaches the optical sensor (14) after passing through said tube (16).The device further comprises a converter (10) arranged to receive one or more light transmission measurements from the optical sensor (14) and to determine a sedimentation rate. Fig.1.

Description

Title of the invention: Device for spreading or coloring and determining sedimentation rate

[0001] The invention relates to the field of hematology, and more particularly to the determination of the sedimentation rate (hereinafter ESR, also called "erythrocyte sedimentation rate" or "ESR" in English).

[0002] The sedimentation rate is part of routine examinations with the complete blood count (hereinafter CBC or NFS for "Completed Blood Count" in English) carried out during a blood test and allows the presence of inflammatory or infectious pathologies to be detected. For example, rheumatism, cancer and other conditions causing changes in the protein concentration in the blood. A high ESR indicates an inflammatory state without prejudging its nature. However, it happens that the ESR remains low while there is an inflammatory syndrome. A high ESR therefore generally indicates the presence of a pathology. The combination of this non-specific test with additional examinations allows for a more precise diagnosis.

[0003] The reference method for measuring ESR is the Westergren method. A narrow tube of standardized dimensions is filled with a blood sample taken with an anticoagulant, for example EDTA, and diluted with sodium citrate and placed vertically. The ESR measurement is the height, in millimeters, of the plasma column after one hour of sedimentation. For example, a height of less than 10 mm after the first hour of sedimentation is considered a normal value for an adult male. The limit of normality is not an absolute value but depends on the age and sex of the patient.

[0004] The ESR is the result of three stages: aggregation of red blood cells, sedimentation, and compaction of the rouleaux.

[0005] The phenomenon of aggregation of red blood cells is produced in particular by blood proteins with a force that is all the greater as the concentrations of the latter are high. The formation of aggregates is the stacking of red blood cells into "rolls", then into three-dimensional structures. The sedimentation rate depends on the dimensions of the aggregates and the viscosity of the plasma.

[0006] During the last stages, the aggregates will sediment, that is to say they will gradually fall to the bottom of the tube, then compact, which will have the effect of separating the sample into a clear translucent portion (plasma), at the top, and a much darker portion (red blood cells) at the bottom. The ESR is measured by the height of the translucent portion.

[0007] The most relevant inflammatory parameter is the aggregation phenomenon which is directly dependent on the concentrations of plasma proteins. It is this phenomenon which is measured by the present invention to return an ESR value. It is also a non-specific parameter, but which eliminates interference resulting from the last two stages of the ESR. There is no standardized parameter allowing this speed or dynamics / kinetics of aggregation to be expressed. It is therefore relevant to transpose aggregation measurement quantities into ESR.

[0008] Due to the quantity of blood (1.6 ml) and the time required to perform it (1 hour), the Westergren method is not compatible with automated hematology analyzers for CBC analysis. To circumvent these problems, research has been conducted to use optical extinction measurements (absorption and diffusion) to determine the sedimentation rate, as in US patent 6,632,679.

[0009] Certain patents, as described in EP 2 921 862 have proposed to integrate a modified ESR measurement into a blood testing device which takes a quantity of blood and separates it into two portions which are respectively subject to a count (NFS) and a measurement of sedimentation rate.

[0010] This device therefore links the counting measurement and that of the sedimentation rate measurement, which is disadvantageous because it requires complex equipment to distribute, via a single sample, two portions of blood samples to the counting section on one side and to the sedimentation rate measurement section on the other, especially since the CBC and ESR measurement operations do not present the same needs in terms of sample preparation.

[0011] More generally, in automated laboratories of the TLA type ("Total Laboratory Automation" in English), the tube flow is managed in a completely automatic manner, from the most frequent tests to the less frequent ones. The equipment necessary to supply the sample to the measuring cell is by far the most expensive and complex part, and includes: - interconnection to a chain, - rack management, - tube agitation, - tube management, - sampling and transfer to the measuring cell, - rinsing of the sampling device, and - the monitoring and transmission devices for results.

[0012] Also, it results in a very high ratio (greater than 100) between the cost of the measurement system and that of the infrastructure necessary to manage the sample. In comparison, this ratio is much lower (2 to 5) for a CBC / DIF / RET measurement system or an automated spreading or staining system commonly used in a hematology laboratory.

[0013] On the other hand, laboratories processing a large number of tubes daily require that the processing time of newly arrived tubes be limited (in English "Turn around time" or TAT) and this directly constrains the sizing of the CBC / DIF / RET measuring systems, and therefore their material cost.

[0014] On the other hand, as a small proportion of the tubes treated require spreading or coloring (only 5 to 20%), this system is commonly oversized by all manufacturers.

[0015] No known device makes it possible to efficiently perform a determination of the sedimentation rate measurement in a rapid and integrated manner in the context of the requirements of a modern laboratory.

[0016] The invention improves the situation. To this end, it proposes a device for spreading or staining and determining the sedimentation rate which comprises a first group arranged to take a blood sample in a tube and to carry out a smear on this sample, and a second group arranged to take a blood sample in a tube and to carry out a determination of the sedimentation rate.The device comprises at least one sampling member controllable for operation by the first group and operation by the second group for sampling a blood sample such that a sample taken for the first group is not used by the second group, and a sample taken for the second group is not used by the first group, the second group being provided with a sensor comprising an infrared light source and an optical sensor arranged substantially opposite each other around a tube connected to an outlet end of the at least one sampling member such that light emitted by the infrared light source reaches the optical sensor after passing through said tube. The device further comprises a converter arranged to receive one or more light transmission measurements from the optical sensor and to determine a sedimentation rate.

[0017] This device is particularly advantageous because it allows the measurement of sedimentation rate to be carried out independently of the blood count. Thus, the measurement of sedimentation rate, which remains a less systematic test, does not interfere with the architecture of the device in a way that could penalize the other functionalities.

[0018] According to various embodiments, the invention may have one or more of the following characteristics:

[0019] - the infrared light source and the optical sensor are arranged at a distance from the sampling end of the sampling organ less than 10 cm, - the second group is placed at an input of the device upstream of the first group, or at the output of the device for carrying out additional measurements of the spreading or coloring type downstream of the first group, - the second group is arranged to carry out rinsing of the sampling organ and the tube between two determinations of sedimentation rate measurements, - the optical sensor is arranged to carry out a blank measurement during a rinsing operation, and in which the converter is arranged to determine the sedimentation rate from the ratio between the blank measurement and the one or more light transmission measurements, - the converter is arranged to determine a lowest light transmission measurement instant, and a final light transmission measurement instant, - the optical sensor is arranged to implement a maximum gain between the lowest light transmission measurement instant and the final light transmission measurement instant, and to implement a minimum gain the rest of the time, - the converter is arranged to calculate the sedimentation rate from the ratio between, on the one hand, the ratio between the blank measurement and the measurement at the final transmission measurement instant, and on the other hand, the ratio between the blank measurement and the measurement at the lowest light transmission measurement instant, - the optical sensor is controlled with a low gain before blood passes into the substantially transparent portion, and with a high gain afterward, and - the sampling organ is a needle which can be controlled for the collection of a blood sample to which is connected a tube in which the substantially transparent portion is made.

[0020] The invention also relates to a method for spreading or coloring and determining sedimentation speed, characterized in that it comprises the use of a device according to the invention, and in that carrying out a spreading or coloring on the one hand and determining a sedimentation speed on the other hand comprise taking two separate samples.

[0021] Other characteristics and advantages of the invention will appear more clearly on reading the following description, taken from examples given for illustrative and non-limiting purposes, taken from the drawings in which: - [Fig.l] represents a general schematic view of a device according to the invention, - [Fig.2] represents a detail of the embodiment of an element of [Fig.l], - [Fig.3] represents a measurement diagram by the device of [Fig.l], and - [Fig.4] represents a schematic view of an embodiment of the device of [Fig.l].

[0022] The drawings and the description below contain, for the most part, elements of a certain character. They may therefore not only serve to better understand the present invention, but also contribute to its definition, where appropriate.

[0023] [Fig. 1] represents a general schematic view of a device 2 for counting blood formula and determining the measurement of sedimentation rate according to the invention. The device 2 comprises a first group 4, a second group 6, a sampling member 8 and a converter 10.

[0024] The first group 4 is arranged to carry out a blood smear. Devices carrying out blood smears automatically are very commonly implemented in laboratories having to process a large number of tubes daily. They are generally largely oversized in terms of processing rate with regard to the needs of the laboratory.

[0025] This oversizing is directly linked to the technical complexity of the different stages necessary for the preparation of the slide of each of the devices (sampling, depositing a drop of aliquot on a glass slide, spreading, identification of the slide, vital staining, etc.): the high rate constitutes the only way for manufacturers to optimize the marginal costs.

[0026] Since blood smears are less systematic tests than the CBC, the Applicant discovered that it was interesting to pool the resources of the first group 4 to add a determination of the ESR measurement using group 6.

[0027] The second group 6 is arranged to determine a measurement of VS. It is this second group which is the main subject of the invention. Indeed, as described in the introduction, this test, whether carried out according to the conventional method or by measuring the change in light absorbance, is either tedious or coupled with a CBC measurement, which slows down the entire device.

[0028] The implementation of the second group 6 according to the invention makes it possible to extrapolate a measurement of sedimentation rate by changing the light absorbance in a manner decoupled from any CBC measurement or other blood measurements. For this, the second group 6 is arranged to control the sampling member 8 which is here a needle independently of the first group 4, that is to say that the samples taken by the first group 4 are only used to carry out a blood smear, and the samples taken by the second group 6 are only used for an ESR measurement.

[0029] Thus, as can be seen in [Fig.l], the first group 4 or the second group 6 controls the needle 8 to take a blood sample from a blood tube 11. Then, this sample is brought to the first group 4 to make a blood smear, or to the second group 6 to make a measurement of change in light absorbance. In the latter case, the measurement is transmitted to the converter 10 which returns an ESR signal and / or a VS measurement value.

[0030] [Fig. 2] represents an exemplary embodiment of the second measuring group 6. According to this example, the second measuring group 6 is produced by means of an optical source 12 and an optical sensor 14 which are assembled facing each other around a tube 16 which serves to transport the blood sample taken by the needle 8 from a blood tube 11 towards the interior of the device 2.

[0031] In the example described here, the light source 12 is of the LED type with an infrared wavelength, preferably in the range 700-980 nm and in particular 800 nm, which is an isosbestic point between oxyhemoglobin and deoxyhemoglobin, which makes the measurement insensitive to the level of oxygen saturation of the blood. In the example described here, the optical sensor 14 is of the photodiode type and can be chosen from PMT, PDA, CMOS sensors, etc. In the example described here, the tube 16 is made of Teflon and is connected to the needle 8. Alternatively, the tube 16 can be made of glass or plastic and must be chosen so as to offer good transparency to the wavelength of the light source 12.

[0032] The light source 12 and optical sensor 14 assembly can be seen as a single sensor 20 of the second group 6. As in the example described here, it can be in two parts which assemble to enclose the needle 8 (for example metallic) and the tube 16. Alternatively, the light source 12 and the optical sensor 14 can be made in one piece. Preferably, the sensor 20 is arranged fairly close to the end of the needle 8, less than 10 cm from it, in order to optimize the shearing of the red blood cells. Preferably, this distance is approximately 5 cm to obtain the best results. In a variant, the sensor 20 can be placed directly at the outlet of the needle 8.

[0033] Whatever the configuration, the second group 6 makes it possible to advantageously use the diameter of the needle 8 which is conventionally of the order of 1 mm. Indeed, during a sample collection by the needle 8, the red blood cells undergo successive deformations which are described below and which are represented in [Fig.3].

[0034] Thus, once the needle 8 is immersed in the blood tube 11, the second group 6 controls aspiration by the needle 8 already primed with diluent in order to draw blood. This is achieved by means of automation control, preparation tanks and solenoid valves and syringes known elsewhere and not shown for reasons of simplification. These means cause the displacement of the diluent which occupies the tube 16. At this stage, due to the translucent nature of the tube 16 and the diluent it contains, the sensor 20 measures a constant optical signal.

[0035] Then, the aliquot of blood is moved to the sensor 20 by suction. The red blood cells undergo shearing, which breaks the aggregation. At this stage, the optical signal remains maximal since the blood is not yet facing the sensor 20. In [Fig. 3], this is represented with the reference 30.

[0036] When the blood arrives in the sensor, the optical signal represents the sheared state of the blood. At this stage of blood shearing, the red blood cells have an elongated shape, as visible with reference 31.

[0037] When the suction stops in order to stop the shearing, the optical transmission signal decreases because the red blood cells return to their relaxed biconcave disc shape. In [Fig. 3], this is represented with the reference 32. Then, the optical signal increases according to a pseudo-logarithmic progression. The progressive increase in the optical signal measured by the optical sensor 14 is linked to the progressive aggregation of the free red blood cells. The aggregation is done by the stacking of the red blood cells forming rolls then three-dimensional structures which is represented in [Fig. 3] with the references 34 and 36.

[0038] Beyond about 40s depending on the blood, aggregation is greatly slowed down and sedimentation begins. This start of sedimentation interferes with the measurement of interest which is aggregation and its correlation with the sedimentation rate. After t4 the aliquot of blood is evacuated, for example by moving the needle 8 above a tank. The blood is thus evacuated and the tube 16 is rinsed with diluent, the tube 16 being at the end of this step entirely filled with diluent as at the start of the procedure described above.

[0039] Four moments are represented in [Fig.3]:

[0040] - before time tl, tube 16 is filled with diluent,

[0041] - the instant tl marks the instant at which the blood arrives at the level of the sensor 20 by suction,

[0042] - the instant t2 marks the instant at which the suction ceases,

[0043] - time t3 marks the time from which the red blood cells have resumed their shape relaxed, and the aggregation begins,

[0044] - time t4 marks the end time of the procedure, with the evacuation of the blood which is again replaced by thinner, as before time tl.

[0045] Observation of [Fig. 3] shows that the optical signal is less significant after time t4, when the tube 16 is filled with diluent, than when the tube 16 is filled with blood, which may seem paradoxical. This is explained by the fact that in the example described here the optical sensor 14 is conditioned differently before time t1 and after time t4 of [Fig. 3].

[0046] Thus, before time t1 and after time t4, it is determined that the measurement corresponds to a "white" measurement. For this entire period, the optical sensor 14 is conditioned by the converter 10 with a minimal measurement gain.

[0047] The instant t3 corresponds to the lowest point of the period t1-t4, from which the optical signal becomes increasing, which marks, as described above, the beginning of the phenomenon that the invention aims to measure. For this reason, and since the variation of the optical signal remains low between the instant t0 and the instant t1, in the example described here, the optical sensor 14 is conditioned with a minimum gain until the instant t1, then with a maximum gain between the instant t1 and t4, then again with a minimum gain after the instant t4 for the next measurement.

[0048] This is all the more advantageous since, in the embodiment described here, the converter 10 is arranged to determine a measurement based on the optical density of the signal measured by the optical sensor 14. As a reminder, the optical density is defined by the formula DO(t) = log^KIQ / 1(0 is the measurement of the optical sensor at the instant t and K the maximum gain / minimum gain ratio. More precisely, the converter 10 is arranged to return a sedimentation velocity measurement based on the DO(t3) / DO(t4) ratio. The Applicant has carried out numerous sedimentation velocity measurements using the Westergren reference method which allows the converter 10 to associate the measurements thus calculated with a VS value.

[0049] The Applicant has discovered that it is particularly advantageous to use optical density, which makes it possible not to depend on possible variations in the transmittance measurement. The Applicant has also discovered that the converter 10 can also operate from the ratio I(t4) / I(t3), without going through the optical density in the Beer-Lambert sense and by using 10 differently in the calculation.

[0050] The converter 10 can be implemented in various ways, for example in the form of appropriate computer code executed on one or more processors. By processors, it is meant any processor suitable for the calculations described below. Such a processor can be implemented in any known way, in the form of a microprocessor for a personal computer, laptop, tablet or smartphone, a dedicated chip of the FPGA or SoC type, a computing resource on a grid or in the cloud, a cluster of graphics processing units (GPUs), a microcontroller, or any other form suitable for providing the computing power necessary for the implementation described below. One or more of these elements can also be implemented in the form of specialized electronic circuits such as an ASIC. A combination of processor and electronic circuits can also be envisaged.In the case of the gradient boosting machine learning unit, dedicated machine learning processors could also be considered. Alternatively, the . converter 10 can be an analog calculator without any programming or computer code per se.

[0051] Still as a variant, the converter 10 could use a machine learning algorithm, involving or not a neural network (deep or not). This involves associating the intensity measurements of the optical sensor 14 with a sedimentation rate value. This variant can be particularly useful for doing without optical density. Still as a variant, the gain of the optical sensor 14 could be the same for all times of the measurement.

[0052] [Fig.4] represents a schematic view of an embodiment of the device of [Fig.l]. As can be seen in this figure, the device 2 is a conventional hematology apparatus comprising a syringe 40 of diameter 1.5mm, and a syringe 42 of diameter 16mm which are both connected to the needle 8. The sensor 20 has been integrated on the needle 8, so that it covers the end of the tube 16 connected to the needle 8.

[0053] The device 2 of [Fig.4] is suitable for carrying out a smear, an ESR measurement or both.

[0054] To carry out a smear by the first group 4, the entire circuit is primed with diluent, then the needle 8 hits the blood tube 11 and takes approximately 75 μl with the syringe. As it rises, the outside of the needle 8 is progressively rinsed by a flow of liquid pushed on one side and continuously sucked and evacuated to the waste on the other.

[0055] The syringe then withdraws the liquid column in order to transfer the aliquot to the Y of an open clamp valve 44 on the side of the needle 8. The clamp valve is activated to close the channel coming from the needle 8 and open the channel to a dropper needle 46. The syringe then pushes which allows the sample to be transferred to the dropper needle. A sequence of peripheral rinsing of the dropper needle and then drying ensures the quality of the initial aliquot front. Finally, a drop of a few microliters is deposited on a slide 48 by pushing the syringe.

[0056] For the ESR measurement, the procedure is as described above with reference to Figures 1 to 3. The entire circuit is primed with diluent drawn from a reservoir 44, then an air bubble of a few pL is created at the end of the needle 8. The needle 8 is lowered into the blood tube 11 and an aliquot of 50 pL to 100 pL is taken by the syringe 42. The needle 8 is raised, and the aliquot transferred to the sensor 20 and the optical measurement of the aggregation is carried out. Finally, the internal and external rinsing of the needle 8 in a tank is carried out, with disposal in the waste. The blank measurement (10) can be carried out before the sample is taken or at the end of rinsing. The optional comparison of the before and after white measurements allow for exception management, for example rinsing control.

[0057] The smear and ESR cycles are carried out independently of each other, and in particular are the subject of a separate sample taken by needle 8: the aliquot for the smear cannot be used for the ESR measurement and vice versa.

[0058] This makes it possible to decouple the two operations without problematic blood sampling when both measurements are requested. This independence makes the ESR measurement much less disruptive to the throughput of the device 2 and allows integration at lower cost, both materially and humanly.

[0059] The device of the invention is much more efficient than all known systems, which offer either a specific module for measuring the sedimentation rate, in addition to the spreading or coloring module, or to tap the CBC / DIF / RET treatment time on a general module.

[0060] Alternatively, the second group 6 can be integrated on the sampling needle of a high-end device which includes a specific sampling module for subsequent spreading or coloring measurements.

[0061] The invention therefore makes it possible to efficiently integrate the measurement of sedimentation rate into existing devices, without impacting their work rates or their architecture.

Claims

Claims

1. A device for spreading or staining and for determining sedimentation rate, comprising a first group (4) arranged to take a blood sample from a tube and to perform a smear on this sample, characterized in that it comprises a second group (6) arranged to take a blood sample from a tube and to perform a determination of sedimentation rate, the device comprising at least one sampling member (8) capable of being controlled for an operation by the first group (4) and an operation by the second group (6) for the collection of a blood sample so that a sample taken for the first group (4) is not used by the second group (6), and a sample taken for the second group (6) is not used by the first group (4),the second group (6) being provided with a sensor (20) comprising an infrared light source (12) and an optical sensor (14) arranged substantially opposite each other around a tube (16) connected to an outlet end of the at least one sampling member (8) so that the light emitted by the infrared light source (12) reaches the optical sensor (14) after passing through said tube (16), the second group (6) being further arranged to carry out a rinsing of the sampling member (8) and of the tube (16) between two determinations of sedimentation rate measurements and the optical sensor (14) being arranged to carry out a blank measurement after a rinsing operation,the device further comprising a converter (10) arranged to receive a white measurement and one or more light transmission measurements from the optical sensor (14) and to determine a sedimentation rate from the ratio between the white measurement and the one or more light transmission measurements.,

2. Device according to claim 1, wherein the infrared light source (12) and the optical sensor (14) are arranged at a distance from the sampling end of the sampling member (8) of less than 10 cm.

3. Device according to one of the preceding claims, in which the second group (6) is arranged at an inlet of the device (2) upstream of the first group (4), or at the outlet of the device (2) for the performance of additional spreading or coloring type measurements downstream of the first group (4).

4. Device according to one of the preceding claims, wherein the converter (10) is arranged to determine a lowest light transmission measurement instant (t3), and a final light transmission measurement instant (t4).

5. Device according to claim 4, wherein the optical sensor (14) is arranged to implement a maximum gain between the lowest light transmission measurement instant (t3) and the final light transmission measurement instant (t4), and to implement a minimum gain the rest of the time.

6. Device according to claim 4 or 5, in which the converter (10) is arranged to calculate the sedimentation rate from the ratio between on the one hand the ratio between the white measurement and the measurement at the final transmission measurement instant (t4), and on the other hand the ratio between the white measurement and the measurement at the lowest light transmission measurement instant (t3).

7. Device according to one of claims 1 to 4, in which the optical sensor (14) is controlled with a first gain before blood passes into the substantially transparent portion, and with a second gain after, the second gain being greater than the first gain.

8. Device according to one of the preceding claims, in which the sampling member is a needle (8) which can be controlled for the sampling of a blood sample to which is connected a tube (16) in which the substantially transparent portion (16) is made.

9. Method for spreading or coloring and determining sedimentation rate, characterized in that it comprises the use of a device according to one of claims 1 to 8, and in that carrying out a spreading or coloring on the one hand and determining a sedimentation rate on the other hand comprise taking two separate samples.

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