JP2022019626A - System and method for determining parameter relevant to health condition of liver of subject - Google Patents

Abstract

To provide a system and a method for determining a parameter relevant to a health condition of the liver of a subject.SOLUTION: A system (1) includes: an elastography device (2); a blood-test reader (3); and a relevant blood-test disposable (10). The disposable includes a capillary tube, and includes reagents appropriate to detect at least one liver enzyme in a blood sample. The blood-test reader is operatively connected to the elastography device, and a control and processing system (4) are configured to control the elastography device and the blood-test reader, and to determine a parameter relevant to a health condition of the liver of a subject, taking account of both a value of at least one mechanical parameter measured by the elastography device and a value of concentration of the at least one liver enzyme.SELECTED DRAWING: Figure 1B

Description

The techniques of the present disclosure relate to systems and methods for determining parameters related to a subject's liver health, particularly based on both liver stiffness measurements and the concentration of liver enzymes in the subject's blood.

Liver hardness as measured by vibration-controlled transient elastography has been shown to be an efficient tool for diagnosing chronic liver diseases such as fibrosis. For example, "Transient elastography: a new non-invasive method for assistance fibrosis", Ultrasound in Medicine and Body, Volume 29, Volume 29, Vol. A paper by Sandrin et al. Shows that liver hardness correlates well with liver fibrosis. However, liver hardness is affected by other factors such as inflammation and congestion.

Interestingly, hepatitis can be assessed by measuring the concentration of one or more liver enzymes in the blood (because hepatitis usually leads to high levels of those enzymes). Therefore, "Live stiffness: a novel parameter for the disease of liver diagnosis", Hepatic Medicine: Evidence and Research, vol. 2, 49-67, 2010, S.A. Muller and L.M. Knowing the concentration of one or more liver enzymes in the blood, as shown in the paper by Sandrin, allows for a better interpretation of liver hardness measurements, potential inflammation / congestion and potential. To be able to obtain reliable information on both fibrosis / cirrhosis.

In order to take these two parameters (serological and mechanical) into account when determining the parameters related to the health of the subject's liver, both liver hardness and liver enzyme concentration should be taken into account. Dependent, the values of benchmark parameters specifically designed to allow identification of various health conditions (or medical conditions) of the subject's liver may be calculated. Such benchmark parameters may be referred to as "scores" in medical works. For example, "FibroScan-AST (FAST) score for the non-invasive identification of patients with non-alcoholic steatohepatitis with significant activity and fibrosis: a prospective derivation and global validation study", The Lancet, Gastroenterology and Hepatology, Volume 5, issue 4 , Pp. 362-373, April 1, 2020, P.M. The paper by Newsome et al. Takes into account the liver hardness "LSM", the ultrasonic attenuation parameter called "CAP" (Control Parameter), and the concentration of aspartate aminotransferase (AST) in the blood "FAST". The benchmark parameters called are shown. This benchmark parameter is calculated by the following equation F1 (where AST is expressed in IU / L):

In any case, in order to detect and characterize a subject's potential liver disease by taking into account both liver hardness and the concentration of one or more liver enzymes, first use those two amounts. Must be measured.

Liver hardness is usually measured by vibration-controlled transient elastography. Such measurements are completely non-invasive and are usually performed in a medical imaging lab environment. In addition, a trained medical professional to take a biological sample takes a venous blood sample using a syringe to determine the concentration of one or more liver enzymes in the subject's blood, and this blood sample Send the sample to the biomedical analysis laboratory to be analyzed. Once both liver hardness and liver enzyme concentration values are available, parameters related to the subject's liver health can be evaluated, for example by calculating benchmark parameters as shown above. ..

However, this processing method has some drawbacks.

First, with respect to test staff and the environment, the requirements for such venous blood-based serological analysis and liver hardness measurements differ significantly. Therefore, those two tests are generally performed in different locations or environments (and thus at different times), which complicates the overall procedure, is more costly, and transfers partial results of the test. Increases the likelihood of error.

Moreover, in this procedure, blood analysis results are not immediately available and a complete diagnosis can only be realized posteriorly (a posteriori). Therefore, when the combined test results (both physical and serological) indicate, for example, an abnormally high value of the FAST parameter and ultimately a potential liver disease, the medical professional will diagnose. You are no longer in a position to repeat one or both of these tests to confirm, which may increase the false positive detection rate.

Also, while this kind of independence of blood analysis for hardness and / or ultrasonic attenuation measurements leads to a systematic analysis of the subject's blood, such blood tests are wasted in some situations. For example, when the negative predictive value (NPV) of liver hardness is excellent, a very low liver hardness, usually less than 6-7 kPa, is present regardless of the concentration of liver enzymes in the blood of the subject (above). (As explained in the paper by Muller and Sandrin referred to in), it is shown that the subject under test is unlikely to have any fibrosis. Therefore, in such situations, testing the subject's blood as is normally done is a waste of resources.

Collecting venous blood allows a very large amount of blood to be freely used, thereby allowing precise and accurate blood analysis. However, instead, it increases the delay (and possibly travel time) between blood data collection and its analysis. Also, this delay will vary significantly from day to day between hospitals or medical testing centers, and even within the same hospital or medical testing center. Also, the chemical activity of liver enzymes in blood samples decreases significantly over time once the sample is taken. Therefore, in a given blood sample, the actually measured concentration of a given liver enzyme (ie, the apparent concentration of this enzyme) varies strongly with the delay between collection and analysis. Therefore, the variability of the delay from collection to analysis leads to the variability of the liver enzyme itself, which is one of the reasons for the normal value indication as well as the measured value.

Furthermore, in this measurement process, blood sampling and liver hardness measurement may be performed at significantly different time points. The lack of this adjustment may also affect the reproducibility of the final diagnosis, as the concentration of liver enzymes in the subject's blood changes over time due to circadian and other variability.

In this regard, blood samples can be analyzed in-situ and there is no need to send the samples to a central laboratory for testing at Point of Care (POC). Blood test devices called devices are known. Some of those POC devices, such as the Piccolo Xpress model by Avaxis (Piccolo Xpress is a registered trademark), provide sufficiently accurate results for further benchmarking and scoring. When using this device, about 100 microliters of blood sample is taken from the subject and then injected into a disc-shaped disposable using a micropipette to test the subject's blood. The disposable is then inserted into a blood test reader, where the disposable is rotated to centrifuge the blood sample in order to separate the cellular components of the blood from the other blood components. The sample then reacts with reagents contained in the disposable that are suitable for optically detecting various blood components such as liver enzymes.

Using a POC device instead of sending a blood sample to a central laboratory for testing makes it possible to improve the process described above.

However, this POC device requires a fairly large amount of blood (~ 0.1 mL). Therefore, the problems associated with collecting such blood samples are exactly the same as those with venous blood collection, and this collection often requires the intervention of a medical professional (especially the collected blood samples are disposable). Very difficult to inject into).

In any case, even if the measurement of the concentration of liver enzyme in the blood of the subject is made with the POC device, the serological and hardness measurements remain separate from each other and therefore the result of this treatment is It still suffers from the variations and variations mentioned above.

"Transient elastography: a new non-invasive method for assistance fibrosis", Ultrasound in Medicine and Body, Volume 29, Volume 29. Written by Sandrin et al. "Live stiffness: a novel parameter for the disease of liver diagnosis", Hepatic Medicine: Evidence and Research, vol. 2, 49-67, 2010, S.A. Muller and L.M. by sandrin "FibroScan-AST (FAST) score for the non-invasive identification of patients with non-alcoholic steatohepatitis with significant activity and fibrosis: a prospective derivation and global validation study", The Lancet, Gastroenterology and Hepatology, Volume 5, issue 4,362 -373, April 1, 2020, P.M. Written by Newsome et al. "Development of a simple non-invasive index to predict significant fibrosis patients with HIV / HCV co-infection", Starling R. et al. K. Etc., Hepatology 2006, vol. 43, 1317-1325

Therefore, it is desirable to further improve the process for determining parameters related to the subject's liver health based on both serological and hardness measurements. In particular, it is desirable to further improve their reproducibility in order to allow better control of test conditions and treatments, simplifying them and reducing associated costs.

In order to at least partially solve the problems described above, the techniques of the present disclosure are systems and methods for determining parameters related to a subject's liver health.
-In this method and system, the blood test reader is operably connected to elastography devices, for example allowing coordinated operation of those devices, for example simultaneous operation.
-In this method and system, the blood test disposables associated with blood test readers are configured to allow non-professional individuals to conveniently and easily collect capillary blood samples.

Specifically, the technique of the present disclosure is a system for determining parameters related to the health condition of a subject's liver:
-With an elastography device configured to measure at least one mechanical parameter associated with shear wave propagation in the liver.
-A blood test reader, and a blood test disposable associated with a blood test reader, the blood test disposable is configured to receive a capillary blood sample and be inserted into the blood test reader, the blood test reader. Includes a blood test reader and blood test disposable-a control and processing system that includes at least a processor and memory, which is configured to determine the concentration of at least one hepatic enzyme in a blood sample.
-Blood test disposable is
-Capillaries for collecting blood samples from fingers,
-Contains reagents suitable for detecting at least one liver enzyme in a blood sample.
-The blood test reader is operably connected to the elastography device
The control and processing system has the following steps:
-S1-Obtaining the value of at least one mechanical parameter measured by an elastography device,
-S2-Obtaining the value of the concentration of at least one liver enzyme in a blood sample taken from a subject as measured by a blood test reader,
-S3- Taking into account both the value of at least one mechanical parameter and the value of the concentration of at least one liver enzyme, the parameters related to the liver health of the subject were determined, and the parameters related to the health condition were determined. It is programmed to output and perform data that represents
The health-related parameters are the health or disease stages identified among the various stages of a given health classification, or the value of at least one mechanical parameter and the concentration of at least one liver enzyme. With respect to the system, which is a benchmark parameter that takes into account both the value of, or is represented by both the health stage or the illness stage and the value of the benchmark parameter.

The control and processing system is configured to control the elastography device and blood test reader. In particular, it is such that the measurement of at least one mechanical parameter and the measurement of the value of said concentration follow a predefined time sequence (eg, within a given time frame) and / or conditionally. It may be configured to control the elastography device and blood test reader as performed according to the measurement procedure (procedure as shown in FIGS. 11 to 15).

Collecting blood samples using capillaries, especially small amounts of capillary blood samples, is easily feasible even for non-medical professionals trained to collect biological samples. In addition, since the capillaries are disposablely integrated, careful injection of the collected blood sample into the previously described disc-shaped disposable is avoided.

Thanks to the specific structure of the blood test disposables described above, medical professionals specially trained to take biosamples and a suitable environment for biosampling are no longer needed. Therefore, thanks to this particular disposable (and even associated reader), the entire test procedure is thus a medical imaging test trained medical professional (two different medical professionals with different specialties). Achieved within a single test environment (under less stringent hygienic constraints than the environment for traditional biosampling), either by (without) or even by a single operator, such as the subject being tested. It is possible to be done. This simplifies the overall procedure and, more importantly, is very helpful in achieving simultaneous measurements of liver hardness and liver enzyme levels, providing reproducibility and certainty of the procedure, as detailed above. Significant improvement.

It is rather unusual to collect a small amount of capillary blood to measure the concentration of liver enzymes, especially in view of determining parameters related to the health of the subject's liver, especially by calculating benchmark parameters. The applicant emphasizes that it is an approach.

In fact, determining parameters related to the subject's liver health and calculating benchmark parameters requires accurate measurements of such concentrations, which is very much of such enzymes in the blood. Difficult due to low concentration. This is why venous blood samples, which allow extensive blood analysis, are usually collected and used for such decisions.

Surprisingly, however, liver enzyme concentrations can be determined very accurately from a limited amount of capillary blood samples, and the accuracy of the results thus obtained is, in fact, such a liver diagnosis. It turns out to be appropriate for. In fact, the use of such a collection method makes it possible to initiate blood analysis immediately after collection, thereby reducing the liver enzyme chemical activity that occurs between collection and analysis in conventional venous blood analysis methods ( Obvious decrease in concentration) is avoided. Therefore, the relative accuracy of the measurements when using a capillary blood sample may be lower than when using a venous blood sample, but the enzyme activity is higher, ultimately related to the health of the subject's liver. It leads to appropriate measurement accuracy to determine the parameters to be used.

It should be noted that the expression "liver enzyme concentration" may indicate a clear concentration of this enzyme as determined using a given detection reaction or set of reactions, or the chemical activity of the liver enzyme in question. You will understand that there is. In this document, the terms "enzyme concentration," "enzyme chemical activity," or "enzyme activity" are used interchangeably.

To allow accurate determination of the activity of one or more liver enzymes from a small amount of capillary blood, the inventor has developed a specially designed blood test disposable and reader. In particular, in order to reduce the amount of blood required to perform this analysis, only determination of liver enzyme activity (instead of the general purpose blood test disposable, which is configured to detect many types of enzymes or blood components) is made. Specific blood test disposables configured to enable it have been developed. This particular blood test disposable may be specifically configured to allow determination of AST and / or ALT activity only.

Moreover, as mentioned above, the fact that the blood test reader and elastography device are operably connected to each other allows for coordinated operation, especially simultaneous operation, of the two devices (more generally). Allows the operation of those two devices, realized according to a predefined time sequence or a predefined conditional measurement procedure). Also, achieving elastographic measurements and corresponding blood tests simultaneously or within a given limited time frame is a health-related parameter or other ultimately obtained, as described in detail above. Significantly improves the reproducibility and certainty of the results. In some embodiments, the parameter related to the subject's liver health may be this health itself (may directly indicate whether or not the liver is considered healthy with respect to some criterion). It may also be an intermediate parameter combined with other information about the subject's liver health or about the subject himself to characterize the liver condition.

Thanks to the above connections, the control and processing system, especially within a single time frame, the elastography device and the blood test reader, blood tests, including elastography measurement processing and blood sample collection and analysis, respectively. Controlling the elastography device and the blood test reader to initiate the process contributes to obtaining simultaneous or near simultaneous measurements (ie, simultaneous or near simultaneous measurements). The control and processing system may be programmed, in particular, so that the timeframe has a duration of up to 30 minutes.

In one embodiment, the elastographic device may be configured, for example, to encourage the operator to implement elastographic measurements of mechanical parameters when initiating the measurement process in question. The blood test reader may also be configured to encourage the operator to take a capillary blood sample from the subject when initiating a measurement process triggered by a control and processing system. This reminder can be communicated to the operator in a variety of ways, such as displaying information on the screen, turning on the indicator light, or opening the blood test disposable dispenser.

The control and processing system also manipulates both taking a capillary blood sample from the subject and achieving an elastographic measurement within a given time when the measurement process in question begins. It can be programmed to convey information that prompts a person (such prompting can be achieved, for example, by displaying a timer or time bar).

The control and processing system may also be programmed to check if elastographic and liver enzyme measurements were completed at the same time. For this purpose, it is examined whether the time gap between the measurement of at least one mechanical parameter and the measurement of the concentration of at least one liver enzyme in a blood sample exceeds a given duration threshold. The control unit may be programmed as such. In addition, the control and processing system relates to an error message indicating that the determination of parameters related to the subject's liver health failed when the above two measurements were not simultaneous, or to the subject's liver health. It may be programmed to output an error message stating that the parameters to be used may not be reliable.

In addition, because the blood test reader and elastography device are operably connected (instead of operating independently of each other), the system dynamically adapts the procedure according to the measurement results. It is possible to carry out diagnostic procedures and workflows in which the two devices interact with each other.

For example, in one embodiment, the control and processing system has the following steps:
-S10-Controlling the elastography device so that the elastography device initiates the elastography measurement process, and then-S1 and then-at least one mechanical parameter value acquired in step S1 is given. If the criteria are met:
-S3'-Regardless of the concentration of at least one liver enzyme in the subject's blood, taking into account the values of at least one mechanical parameter, determine the parameters related to the subject's liver health, said parameters. Outputting data representing, on the other hand
-If the value of at least one mechanical parameter does not meet the criteria:
-S20-Information clearly stating that a blood test is recommended for the subject's liver health assessment, and / or for the operator to take a capillary blood sample from the subject and begin analysis of this blood sample. It may be programmed to control the operator interface so that the operator interface sends prompting information, followed by-step S2 and then-step S3.

Thanks to these features, blood tests are performed only when it is expected to significantly improve the determination of parameters related to the subject's liver health, thereby saving time and resources.

In particular, the control and processing system will only test the blood if the value of at least one mechanical parameter obtained in step S1 indicates that the subject's liver hardness exceeds a given threshold, eg, 5-6 kPa. Can be programmed to control the blood test reader to start.

More generally, the control and processing system sets a threshold corresponding to the boundary between, on average, a value at which the liver does not suffer from health problems and a value at which the liver may have health problems. When the value of one mechanical parameter is exceeded, it may be programmed to determine that the value of at least one mechanical parameter does not meet the criteria (hence a blood test is recommended).

In other embodiments, a system for determining parameters related to a subject's liver health first initiates a liver enzyme measurement process and then (to determine if liver enzyme measurement is useful). It may be configured to start the elastography measurement process (instead of starting the elastography measurement).

Even with a blood test disposable and blood test reader optimized to allow rapid determination of liver enzyme levels, the determination is not immediate and usually takes several minutes (this duration is The detection reagent is required to react with the liver enzyme). Therefore, it is beneficial to start the liver enzyme measurement process as soon as possible and use the time required for blood sample analysis to make the elastographic measurement. In fact, proceeding in this order reduces the time required to complete the entire test procedure.

In this last embodiment, the control and processing system specifically:
-S20'-Once a capillary blood sample is taken from the subject and then the blood test disposable is inserted into the blood test reader, the operator interface sends information prompting the operator to start analyzing this blood sample. To control the operator interface to do
-S10-Controlling the elastography device so that the elastography device initiates the elastography measurement process, and then-steps S1 and S2, and then.
-It may be programmed to perform step S3 and.

In addition, in some embodiments,
-In the memory of the control and processing system, various formulas are associated with different ranges of values of at least one mechanical parameter, and each formula corresponds to a given benchmark parameter for liver health assessment, and each The formula allows the corresponding benchmark parameters to be calculated from at least one mechanical parameter and the concentration of at least one liver enzyme in the blood sample.
-The control and processing system compares the values of at least one mechanical parameter previously measured with respect to the subject's liver to the range of values associated with their various benchmark parameters, respectively, of those benchmark parameters. Programmed to select one of them,
-The control and processing system is programmed in step S3 to calculate the value of the previously selected benchmark parameter according to the equation associated with this benchmark parameter.

Next, with respect to the blood test disposable and related blood test readers, in one embodiment, the blood test disposable is a blood sample taken using a capillary tube at a maximum of 60 microliters, or at a maximum of 40 or 30 microliters. It is configured to have even a quantity.

The amount of capillary blood that can be collected with a single finger stab is typically 10 to 20 microliters. Thus, the blood samples described above can be taken by achieving a limited number of finger stabs, or even a single finger stab. Therefore, there is limited inconvenience to the subject (especially avoiding stinging the subject's fingers altogether). Also, if it is found that the liver enzyme measurement must be done again, another blood sample of this type can be taken again.

In some embodiments, the capillaries are fixed to a portion of the disposable, eg, irremovably fixed.

In particular, blood test disposables are:
-Cartridge containing reagents and
-May include removable plugs
Capillaries are secured to removable plugs or cartridges, and the plugs and cartridges
-The plug can be removed from the cartridge and then reinserted into the cartridge, and-When the plug is inserted into the cartridge, the capillaries are housed inside the disposable and collected by the capillaries. The blood sample is configured to come into contact with the reagent.

Therefore, in order to collect a blood sample, an operator (which may be the subject to be tested himself) pierces the skin of at least one finger of the subject, for example using a lancet. The operator also removes the plug from the cartridge and then uses a capillary tube to collect a small drop of capillary blood thus obtained. After that, the operator reinserts the plug into the cartridge. Therefore, the capillaries remain contained within the disposable for most of the time, which prevents possible contamination of the capillaries, or possible contamination of the capillaries after the blood sample is taken. Moreover, thanks to this structure, the collected blood samples are consistently and easily mixed with the reagents in a reproducible manner, avoiding careful infusion into conventional disposables.

According to the optional feature of the system described above, the plug is configured such that inserting the plug into the cartridge increases the pressure to push the blood sample collected using the capillaries into the fluid circuit. In addition, the cartridge may include a fluid circuit configured to contact at least a portion of the blood sample with the reagent. The plug may also be configured such that the capillaries fluidly connect to this fluid circuit when the plug is plugged into the cartridge.

In one embodiment of the system described above:
-At least one liver enzyme is one of: aspartate aminotransferase (hereinafter "AST"), alanine aminotransferase (hereinafter "ALT"), γ-glutamyl transferase (hereinafter "GGT").
-Reagents are suitable for optically detecting at least one liver enzyme.
-A blood test disposable is configured so that at least a portion of the blood sample mixes with the reagent in the first reaction zone.
-A blood test reader uses at least a light source and an optical sensor to determine the concentration of at least one liver enzyme in a blood sample using light reflectance and / or transmittance measurements in the first reaction zone. And include.

The blood test disposable may include a filter membrane configured to filter red blood cells from a blood sample taken from a subject to obtain a plasma sample, contacting at least a portion of the plasma sample with a reagent. It is configured to let you.

This method of filtration using this membrane does not have to be as complete as other filtration methods (such as centrifugation) and produces plasma samples rather than serum samples, but this filtration is sufficient for subsequent liver enzyme detection. It is efficient, faster, and causes a small amount of loss compared to previous methods (this volume loss is usually less than 60% of the original amount of blood sample collected) and is collected here. It is beneficial because the amount of plasma blood sample is small and the object of the technique of the present disclosure is to reduce the time required to determine the parameters related to the liver health of the subject. ..

In one embodiment of the system described above:
-At least one liver enzyme is AST or ALT,
-Reagents contain α-ketoglutaric acid and L-aspartic acid or L-alanine, respectively.
-The blood test disposable contains the following catalysts: pyruvate oxidase and peroxidase, and contains an indicator that is colored when at least one liver enzyme reacts with the product of the set of reactions that occur when mixed with the reagent.

In particular, the indicator may contain a MAOS Trisder reagent, the blood test disposable may contain a liquid buffer, and the buffer is configured to mix with the reagent when the blood test disposable is inserted into the blood test reader. This buffer is an aqueous solution containing tris (hydroxymethyl) aminomethane (hereinafter, TRIS).

The concentration of TRIS in the buffer may be, in particular, 0.05 to 1 mol / liter, or even 0.1 to 0.5 mol / liter.

The use of this indicator and buffer allows for rapid and accurate determination of the concentration of at least one liver enzyme in the collected blood sample. In fact, when this system is used, measurement results are usually obtained in 10 minutes or less (usually 5 minutes) after blood sampling, and the measurement accuracy is better than 20%, usually about 10 minutes. %, Or even less than about 10%.

The MAOS Trinder reagent represents N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3,5-dimethylaniline sodium salt. Further, tris (hydroxymethyl) aminomethane (TRIS) represents 2-amino-2-hydroxymethylpropane-1,3-diol.

In one embodiment, the above-mentioned reagents are contained in a dryness detection pad, while the catalyst and indicator are contained in a dryness detection substrate separate from the pad, and the substrate and pad are configured in the above-mentioned first reaction zone. By doing so, at least a portion of the plasma sample that may be mixed with the buffer penetrates the detection pad and substrate pad, thereby allowing liver enzyme detection.

The buffers described above may also contain phosphate and / or have a pH between 6.5 and 8, which is suitable for the detection of liver enzymes described above.

In one embodiment:
-Blood test disposable includes on-board control of catalytic activity, on-board control,
-A dry control board containing oxaloacetate and
-Contains a dry control pad containing a catalyst and an indicator, the control pad is separate from the control substrate,
-The blood test disposable is configured so that the liquid penetrates the control board and control pad when the blood test disposable is inserted into the blood test reader or when the blood sample is received in the blood test disposable. ..

In the presence of pyruvate oxidase and peroxidase, oxaloacetate reacts to produce hydrogen peroxide in particular. The MAOS is then oxidized by hydrogen peroxide and colored thereby, which is optically detected by a blood test reader, thereby confirming the integrity and activity of the catalysts (pyruvate oxidase and peroxidase). Therefore, this onboard control improves the certainty of the system (especially when low liver enzyme concentrations are measured, this control confirms that this result is not due to catalytic defects). ..

It should be noted that pyruvate oxidase and peroxidase allow detection of either AST or ALT (when combined with α-ketoglutaric acid and L-aspartate or L-alanine, respectively). Therefore, AST (in the first reaction zone) and ALT (in the first reaction zone) and ALT (in the first reaction zone) in a fairly complete manner, using a single onboard control such as those described above (rather than requiring two separate controls). The integrity of the disposable, which is configured to detect both (in the reaction zone of 2), is evaluable, which simplifies the structure of the disposable and the amount of liquid required for its operation.

The above-mentioned liquid that penetrates the control substrate and control pad when the blood test disposable is inserted into the blood test reader may be a buffer solution contained in a blister that can be broken. The blister may be configured to break and inject buffer into the control pad and substrate when the blood test disposable is inserted into the blood test reader. Using such buffers to achieve this completeness control instead of using plasma samples extracted from collected blood samples makes it possible to reduce the blood volume required for liver enzyme measurements. , Very informative.

In some embodiments, the system is further configured to be able to determine the platelet count in that blood sample or another blood sample taken from the subject, and the control and processing system also takes into account the platelet count. , Step S3 is programmed to determine parameters related to health status.

In some embodiments of the system:
-The elastography device is also configured to measure ultrasonic attenuation parameters in the liver.
-Control and processing system:
-In step S1, the value of the ultrasonic attenuation parameter measured by the elastography device is acquired.
-In step S3, the value of the ultrasonic attenuation parameter is also taken into account and further programmed to determine the parameter related to the health condition of the subject's liver.

In particular, the control and processing system takes into account the value of at least one mechanical parameter, the value of the ultrasonic attenuation parameter, and the value of the concentration of at least one liver enzyme in step S3, and the liver health of the subject. It may be programmed to calculate benchmark parameters that represent the state.

At least one mechanical parameter related to the propagation of shear waves in the liver may be liver hardness, the ultrasonic attenuation parameter may be an ultrasonic attenuation coefficient per unit length, and at least one liver enzyme may be AST. The control and processing system may also be programmed to calculate the benchmark parameters as equal to or proportional to the exponential function of X divided by the exponential function of 1 + X, where X is the logarithm of liver hardness. And the first-order coupling of the ultrasonic attenuation coefficient to the third power, minus the correction term that everything higher than the concentration of AST is small. This correction term may be proportional to, for example, the concentration of 1 / AST. In particular, the benchmark parameters described above FAST calculated according to equation F1 (corresponding to X = -1.65 + 1.07 x ln [LSM] + 2.66 x ^10-8 x CAP ^3-63.3 / AST). It can also be a parameter.

Using such benchmark parameters, especially when AST concentration measurements and liver enzyme measurements are simultaneous, makes it possible to very reliably discriminate between a subject's liver health status and poor / pathological status.

According to the techniques of the present disclosure, it is understood that the various embodiments described above can be combined according to all technically possible combinations.

The technique of the present disclosure also provides a method for determining parameters related to a subject's liver health using the system, the system:
-With an elastography device configured to measure at least one mechanical parameter associated with shear wave propagation in the liver.
-A blood test reader, and a blood test disposable associated with a blood test reader, the blood test disposable is configured to receive a capillary blood sample and be inserted into the blood test reader, the blood test reader. Is a blood test reader and blood test disposable, configured to determine the concentration of at least one liver enzyme in a blood sample.
-Includes a control and processing system that includes at least a processor and memory.
-Blood test disposable is
-Capillaries for collecting blood samples from fingers,
-Contains reagents suitable for detecting at least one liver enzyme in a blood sample.
-The blood test reader is operably connected to the elastography device
The method is as follows:
-S100-For the liver of the subject under test, the value of the mechanical parameter was measured using an elastography device.
-S1-The control and processing system acquires the value of at least one mechanical parameter measured by the elastography device.
-S200- Capillary blood samples were taken from the subject using capillaries, after which the blood test disposable was introduced into the blood test reader.
-S2-The control and processing system obtains the value of the concentration of at least one liver enzyme in the blood sample taken from the subject as measured by the blood test reader.
-S3-The control and processing system takes into account both the value of at least one mechanical parameter and the value of the concentration of at least one liver enzyme to determine the parameters related to the subject's liver health. Outputs data representing parameters related to health status,
Including
The health-related parameters are the health or disease stages identified among the various stages of a given health classification, or the value of at least one mechanical parameter and the concentration of at least one liver enzyme. It is a benchmark parameter that takes into account both the value of, or is represented by both the health stage or illness stage and the value of the benchmark parameter.

In this method, the control and processing system controls the elastography device and blood test reader. In particular, it is that the measurement of the at least one mechanical parameter and the measurement of the value of the concentration follow a predefined time sequence (eg, within a given time frame) and / or conditionally. The elastography device and the blood test reader may be controlled so as to be performed according to the measurement procedure (procedure as shown in FIGS. 11 to 15).

Steps S100 and S200 may be performed, in particular, within a single time frame with the maximum preset duration. This maximum duration may be 30 minutes, or even 15 or 10 minutes.

In the method described above, the control and processing system may output an error message when steps S100 and S200 are not performed within a single time frame with maximum duration.

The characteristics of various embodiments of the system described above may also be applied to the method for determining the parameters related to the liver health of the subject exactly presented.

Other properties and advantages of the techniques of the present disclosure will be apparent from the illustrations given below, exemplary and non-limitingly, with reference to the drawings.

FIG. 5 is a diagram schematically showing a first embodiment of a system for determining parameters related to the health condition of a subject's liver according to the technique of the present disclosure. It is a figure which shows the system of FIG. 1A more concretely. FIG. 5 is a diagram schematically showing a second embodiment of a system for determining parameters related to the health condition of a subject's liver according to the technique of the present disclosure. FIG. 5 is a diagram schematically showing a third embodiment of a system for determining parameters related to the health condition of a subject's liver according to the technique of the present disclosure. FIG. 3 is a schematic perspective view of a blood test disposable that may include any of the systems of FIGS. 1A-3. 5 and 6 are schematic bottom and top views of the blood test disposable of FIG. 5 and 6 are schematic bottom and top views of the blood test disposable of FIG. It is a schematic side view of the detection pad of the blood test disposable of FIG. FIG. 4 is a schematic side view of the blood test disposable onboard control of FIG. FIG. 4 is a diagram showing a chemical reaction associated with liver enzyme detection carried out in the blood test disposable of FIG. 4. It is a flowchart which shows the various steps of the above-mentioned liver enzyme detection. FIG. 6 illustrates the steps of a first embodiment of a method for determining parameters related to a subject's liver health, according to the techniques of the present disclosure, which can be achieved using any of the systems of FIGS. 1A-3. Is. FIG. 6 illustrates the steps of a first embodiment of a method for determining parameters related to a subject's liver health, according to the techniques of the present disclosure, which can be achieved using any of the systems of FIGS. 1A-3. Is. FIG. 11 is a diagram schematically showing how various benchmark parameters are selected and subsequently calculated in order to determine the parameters related to the liver health of the subject in the method of FIG. It is a figure which shows the calculation and inspection sequence performed in one example of realization of the method of FIG. 11 in more detail. FIG. 6 illustrates the steps of a second embodiment of a method for determining parameters related to a subject's liver health, according to the techniques of the present disclosure, which can be achieved using any of the systems of FIGS. 1A-3. Is.

1A, 2 and 3, respectively, show first, second, and third embodiments of System 1; 1'; 1 "for determining parameters related to a subject's liver health. ..

In each of these embodiments, the system 1; 1'; 1 "is:
-Elastography device 2; 2'; 2 ", which is configured to measure at least one mechanical parameter related to the propagation of shear waves in the liver, such as liver hardness.
-Includes a blood test reader 3 for determining the concentration of at least one liver enzyme in a blood sample taken from a subject under test, and a blood test disposable 10 associated with the blood test reader 3.

System 1; 1'; 1 "further controls the elastography device 2; 2'; 2" and the blood test reader 3 when testing the subject, eg, according to the test process shown in FIG. 11 or FIG. Control and processing system 4; 4'; 4 "included.

Control and processing system including at least processor and memory 4; 4'; 4 "is:
-With the value of at least one mechanical parameter measured by the elastography device 2'; 2 ";2" during the subject's test.
-Programmed to determine parameters related to the subject's liver health, taking into account both the value of the concentration of at least one liver enzyme measured by the blood test reader 3 during this test. To.

The main difference between the three embodiments of system 1; 1'and 1 "(shown in FIGS. 1A, 2 and 3, respectively) is that the control and processing system 4; 4'; 4" is system 1; 1'. 1 ”. For example, in the case of FIG. 1A, the control and processing system 4 is housed in the same casing 6 as the elastography device 2, and is substantially in the form of a control unit of the elastography device 2. On the other hand, in the case of FIG. 2, the control and processing system 4'is a separate electronic device outside both the casing 6'of the elastography device 2'and the casing of the corresponding blood test reader 3. Also, in FIG. 3, a part of the control and processing system 4 "is realized using a non-local and remote computing resource 7 (such as" cloud ").

In any case, these three embodiments of System 1; 1'; 1 "have many common features. Therefore, the same or corresponding elements of those different embodiments may be described only once. May be identified by the same reference symbol / number.

In each of these embodiments, the blood test reader 3 is operably connected to the elastography device 2; 2'; 2 ", which connection is directly feasible without an intermediate system. It can also be realized via an intermediate device, as in FIG. 2, where the elastography device 2'is operably connected to the blood test reader 3 via the control and processing system 4'.

In either case, the connection 9 between the elastography device 2; 2'; 2 "and the blood test reader 3 allows for coordinated operation, eg, simultaneous operation, between the two devices.

The connection 9 may be used, for example, to send a command or execution request from the elastography device 2 to the blood test reader 3. In this case, the elastography device 2 and the blood test reader 3 may be programmed to operate according to the master-slave model, and the elastography device 2 becomes the master and gives a command to the blood test reader 3 to perform a blood test. The reader 3 executes them and responds by sending positive response data, measurement results, data regarding the liver enzyme measurement process, and / or data representing the status of the blood test reader 3.

Also, the connection between the elastography device 2'and the blood test reader 3'is not directly determined by the elastography device 2'but is transmitted by the elastography device 2'(liver hardness measurement results or status data, etc.). It may be possible to send a command or execution request determined based on to the blood test reader. In particular, those commands or requests may be determined by the control and processing system 4'based on the data received from the elastography device 2', whereby the control and processing system 4'may control the elastography device 2'. It plays an active role in connecting to the blood test reader 3 (hence, this connection is some complex and positive connection). Conversely, the elastography device 2'may receive a command or execution request determined (by the control and processing system 4') based on the data received from the inspection reader 3.

The connection 9 between the elastography device 2; 2'; 2 "and the blood test reader 3 is wired or wireless according to USB, Wireless, Bluetooth, 6LoWPAN, ZigBee, Z-Wave, Sigfox, or another protocol. It may be realized by using a link.

As mentioned above, the connection 9 between the elastography device 2; 2'; 2 "and the blood test reader 3 allows for coordinated operation, eg, simultaneous operation, of the two devices, as well as control and control. The processing system 4; 4'; 4 "also performs elastography measurement processing and liver enzyme measurement processing in a coordinated manner, particularly simultaneously (possible by the connection described above), respectively. It may be programmed to control the device and the blood test reader.

As mentioned in the overview section above, achieving simultaneous measurement of mechanical parameters such as liver hardness and the concentration of at least one liver enzyme will further determine the parameters related to the health of the subject's liver. It should be more reliable and reproducible.

The specific blood test Disposable 10 and Reader 3 used to measure the concentration of at least one liver enzyme in the subject's blood participate in this improvement. These, in fact, accurately and quickly, in situ, determine the concentration of at least one liver enzyme in the subject's blood from a very limited amount of capillary blood taken from the subject. It is possible to do. These characteristics, especially the fact that the amount of blood that needs to be collected, is very helpful in simplifying the entire procedure and achieving simultaneous measurements of liver hardness and liver enzyme levels.

In the following, the overall structure of system 1; 1'; 1 "is shown in more detail with reference to FIGS. 1A to 3. This system 1; 1'; 1 with reference to FIGS. 4 to 10. The specific blood test disposable 10 and reader 3 used in "" are further presented. Also, with reference to FIGS. 11 and 15, respectively, two embodiments of methods for determining parameters related to the health of the subject's liver are presented. Each of those two methods is feasible using any of the three embodiments of system 1; 1'; 1 "described above, and any of those systems 1; 1'; 1". Control and processing system 4; 4'; 4 "may be programmed to perform one or the other of those methods.

In the system 1 of FIGS. 1A and 1B, the control and processing system 4 is a part of the elastography device 2 and acts as a control unit of the elastography device 2 (in addition, controls the blood test reader 3). ).

As mentioned above, the elastography device 2 is configured to measure at least one mechanical parameter associated with the propagation of shear waves in the liver. Here, this mechanical parameter is a quantity related to liver hardness, such as the propagation velocity Vs of the shear wave, the shear elastic modulus of the liver tissue or its Young's modulus E. This parameter is presented here as liver hardness or LSM (Live Stiffness Measurement). However, in other embodiments, the mechanical parameters described above can also be quantities related to low frequency (eg, less than 500 Hz) shear wave attenuation in the tissue, such as viscosity.

The elastography device 2 may be configured to measure at least one mechanical parameter, for example, by transient elastography.

Here, the elastography device 2 is further configured to measure the ultrasonic attenuation parameter per unit length, which is called the ultrasonic attenuation parameter in the liver, more precisely the control attenuation parameter or CAP. .. This ultrasonic attenuation coefficient represents the attenuation of high frequency ultrasonic waves in the organ under test (these ultrasonic waves usually have a central frequency of several MHz, for example 2 to 5 MHz).

The elastography device 2 includes an elastography module 21 configured to drive the probe 22 and receive a signal acquired by the probe 22.

The probe 22 sends and responds to at least a transducer to generate a shear wave and an ultrasonic shot to track how the shear wave generated by the transducer moves the subject's liver. Includes an ultrasonic transducer and an ultrasonic transducer for receiving an echo signal. The probe 22 is held against the subject's skin during liver hardness measurement.

The elastography module 21 includes an ultrasonic front end that includes an electronic module for generating the transmitted ultrasonic signal and acquiring and preprocessing the ultrasonic echo signal received by the ultrasonic transducer of the probe 22. The elastography module 21 further includes a motion actuator servo controller for driving the oscillator of the probe 22.

The elastography module 21 also has dedicated logic circuits (eg FPGA, ASIC (application specific integrated)) for processing the acquired echo signals in order to derive the values of the measured mechanical parameters (eg liver hardness). Circuits), or other types of programmable microcircuits). Further, this dedicated logic circuit may be included in the control and processing system 4 instead of being included in the elastography module 21.

The control and processing system 4 is connected to the elastography module 21 and an operator interface 5 such as a display screen. During the measurement of the mechanical parameters described above, the elastography module 21 and the operator interface 5 are controlled by the control and processing system 4. For example, the control and processing system 4 displays guidance information that helps the operator position the probe in front of the subject's liver and obtains measurement results (such as the final elastogram and hardness value). The operator interface 5 is controlled so as to be displayed.

As already mentioned, the control and processing system 4 includes at least a processor and memory coupled to the processor. More generally, it includes electrical circuits for processing data and for transmitting and receiving data. The memory includes a physical non-temporary (nonvolatile) memory module for storing machine executable instructions executed by the processor to perform the functions of the control and processing system 4. It may also include RAM memory for storing signal data and instructions during system operation.

In system 1 of FIG. 1A, the control and processing system 4 and the elastography module 21 are both housed in the casing 6 of the elastography device 2. The operator interface 5 can be integrated into this casing 6 or can be realized in the form of a separate remote device. Even if the elastography device 2 and the blood test reader 3 are placed in the same room to avoid moving the subject or blood sample from one place to another in order to avoid travel time or error. Well, or less than 15 meters may be separated from each other.

As already mentioned, the control and processing system 4 is also operably connected to the blood test reader 3 through the connection 9 described above. Also, the control and processing system 4 is not only programmed to manage the execution of the above-mentioned mechanical parameter measurement processing by the elastography device 2. It is also programmed to control the blood test reader 3 in conjunction with mechanical parameter measurements, eg, to trigger a liver enzyme measurement process at the same time. For example, the control and processing system 4 may be programmed to initiate a liver enzyme measurement process according to the initially obtained liver hardness value (liver enzyme measurement process immediately or nearly immediately after this hardness value is obtained). Can be started). Also, the liver enzyme measurement process and the mechanical parameter measurement process may be programmed to start in close parallel with only a short delay (eg, less than 5 minutes or less than 15 minutes) between the start of each.

The control and processing system 4 also, after the ex post facto, between the measurement of the mechanical parameters described above and the measurement of the concentration of one or more liver enzymes in the subject's sample, once those measurement processes are completed. It may be programmed to check that the time gap is less than a given duration threshold. If this time gap exceeds this duration threshold, the control and processing system 4 may have failed to determine a parameter related to the subject's liver health, or this parameter may not be certain. / Or may output an error message stating that this health determination or transmission may be skipped.

In either case, as already mentioned, the control and processing system 4 has the above-mentioned mechanical parameter values measured by the elastography device 2 and one or more liver enzymes measured by the blood test reader 3. It is programmed to determine parameters related to the subject's liver health, taking into account both the concentration value (eg, based on the results of prior hardness measurements, depending on some conditions). It is the same whether or not it is done).

In describing the methods of FIGS. 11 and 15, various methods for determining parameters related to a subject's liver health are presented from these two measurements, and perhaps from other measurements as well.

The control and processing system 4 of system 1 of FIG. 1A may be specifically programmed to control the blood test reader 3 and the elastography device 2 according to the method of FIG. 11 or the method of FIG. .. In particular, the control and processing system 4 may be programmed to perform steps S10, S1, S20, S2, and then S3 (or S3') of the method of FIG. Alternatively, it may be programmed to perform steps S20, S10, S1, S2, and then S3 of the method of FIG.

The system 1'of FIG. 2 is similar to that of FIG. 1A, but in this second embodiment, the control and processing system 4'is separate from the elastography device 2'.

The elastography device 2'includes the above-mentioned elastography module 21 and the probe 22. The elastography module 21 is housed in a casing 6'separate from the casing 41' of the control and processing system 4'to which it is connected. An operator interface 5'similar to the operator interface 5 described above is connected to the control and processing system 4'.

In the second embodiment, the control and processing system 4'is an electronic personal computer or smartphone, such as a notebook or tablet computer, operably connected to both the elastography device 2'and the blood test reader 3. It may be a logic circuit or system (with one or more processors and one or more memories).

The blood test reader 3 and the disposable 10 are the same as those of the system 1 of FIG. 1A, and the blood test reader 3 is connected to the control and processing system 4'as in the system 1 of FIG. 1A.

Also, an additional optional connection (not shown in FIG. 2) is a blood test directly through the elastography device 2'(specifically, its elastography module 21) (without passing through the control and processing system 4'). It is also possible to link to the reader 3.

The system 1 "of FIG. 3 is similar to that of FIG. 1A, but in this third embodiment, a part of the control and processing system 4" is a non-local remote computational resource 7 (such as a "cloud"). Realized using.

In the third embodiment, the control and processing system 4 "is:
-With a first portion 41 configured to control the "elastography device 2" and the blood test reader 3 and to control the measurement process,
-In particular, it includes a second part 42 configured to determine parameters related to the health of the subject's liver by calculating benchmark parameters (scores).

The first part 41 may be realized locally. Specifically, they are integrated into the elastography device 2 "and housed in the casing 6 of the elastography device 2". On the other hand, a second part 42 of the control and processing system, such as non-local computing services and databases, is remote and is implemented using remote, and perhaps distributed, computing resources 7. It means that at least some of these resources are at least one kilometer away from the elastography device).

The first part 41 and the second part 42 of the control and processing system 4 "are operably connected to each other and thus exchange data and instructions. Also, the blood test reader 3 is (elastography device). It may be directly connected to the second remote portion 42 of the control and processing system 4 "without passing through the first portion 41 (accommodated in), which data processing and data processing the liver enzyme measurement results. It is possible to transfer directly to the second part 42 of the control and processing system 4 "which is more specialized in arithmetic.

After the second part 42 of the control and processing system 4 "determines the data representing the parameters related to the liver health of the subject, this data is:
-To the first part 41 of the control and processing system 4 "and / or-remote, such as a personal computer or smartphone, so that this parameter can be transmitted to the operator using the operator interface 5. It can be programmed to send to device 8 and / or to a remote database for storage.

Except for the differences described above, the third embodiment of the system 1 "is the same as, or at least similar to, the first embodiment. In particular, the elastography device 2" is an elastography module in the same casing 6. 21 (connected to the probe 22) and a first portion 41 (control unit) of the control and processing system 4 ". This first portion 41 of the control and processing system 4" is connected by a connection 9. , Connected to a blood test reader 3 (which may be the same as that of FIGS. 1A and 2).

Next, with reference to FIGS. 4 to 10, the blood test disposable 10 of the system 1 of FIG. 1A is shown in detail. As mentioned above, this blood test disposable 10 can be used without distinction in any of the three embodiments of System 1; 1'; 1 "described above.

The blood test disposable 10 includes a capillary tube 13 for collecting a blood sample from a subject. For this purpose, the skin of the subject's fingers, or perhaps two or three finger skins, is stabbed, for example, with a lancet. Then, the capillary blood droplets thus obtained are collected using capillaries. Here, the capillaries 13 are fixed to a part of the blood test disposable (not a removable and replaceable part), more strictly irremovably fixed.

As shown in FIG. 5, the blood test disposable 10 has two separate parts, namely:
-A cartridge 11 containing a reagent suitable for detecting at least one liver enzyme, and
-Includes a removable plug 12 (in FIG. 5, the plug 12 is shown removed from the cartridge 11).

The capillary tube 13 is fixed to a removable plug 12. However, in other embodiments not shown, the capillaries can also be fixed to the cartridge instead of being fixed to the plug.

The plug 12 and the cartridge 11 are configured so that the plug 12 can be removed from the cartridge 11 and then reinserted into the cartridge 11.

For this purpose, the plug 12 and the cartridge 11 are such as ribs (plug side) and grooves (cartridge side) having complementary shapes, or indentations or holes corresponding to elastic teeth or clips, respectively. It may be provided with male and female fixing elements (or vice versa, female and male elements).

Here, the plug 12 can be removed from the end face 121 of the cartridge, i.e., the rear surface (the front surface of the cartridge is the front surface that is first introduced into the reader 3 when the disposable 10 is introduced into the reader 3). It is a kind of cap that is fixed to.

Capillaries 13 project from the plug 12 to allow blood to be drawn from the subject. It also extends inside the plug 12 to provide a receiving volume large enough to receive blood samples between 20 and 50 microliters, for example 25 +/- 10 microliters. ing.

When the plug 12 is inserted into the cartridge 11 (as in FIG. 6), the outer portion 130 of the capillary tube 13, that is, a part of the capillary tube 13 protruding from the plug 12, enters the cartridge 11. Therefore, the capillaries 13 extend entirely within the blood test disposable 10 (ie, thus the capillaries 13 are entirely contained within the blood test disposable 10).

To collect a blood sample, the operator (which may also be the subject under test) stabs the skin of at least one finger of the subject, for example with a lancet. Further, after removing the plug 12 from the cartridge 11, the operator uses the end 131 of the capillary tube 13 to collect a small drop of capillary blood thus obtained. Then, when the capillary tube 13 (and possibly the auxiliary reservoir) is filled with capillary blood, the operator reinserts the plug 12 into the cartridge 11. In order to collect an appropriate blood volume, that is, here, to fill the capillaries 13, the operator may stab one or more fingers of the subject, for example two or three fingers.

The blood test disposable 10 is configured such that the blood sample collected using the capillary tube 13 in this manner comes into contact with the reagent contained in the cartridge 11.

In the illustrated embodiment, the blood sample collected using the capillary tube 13 exits this tube by the same tube end 131 as that used to collect the blood sample (flows into the cartridge and mixes with the reagent). To do). Further, here, the plug 12 is configured to be inserted into the cartridge 11 to increase the pressure inside the cartridge 11 to push the blood sample contained in the capillaries toward the cartridge 11. For this purpose, the plug 12 may have a variant that is pushed in when the operator pushes the plug 12, for example to insert it into a cartridge.

Upon exiting the capillary 13, the blood sample may be brought into contact with the reagents described above, more generally with a fluid circuit (such as a microfluidic circuit) containing one or more pipes and / or junctions. It may be widely routed through various parts of the cartridge used for enzyme detection and control. It may also reach these reagents directly or by penetrating and traversing one or more membranes or substrates (without going through pipes or other circuits).

As shown in FIG. 6, when the plug 12 is inserted into the cartridge 11, the output end 131 of the capillary tube 13 is arranged directly above the filtration membrane 110. The filtration membrane 110 is suitable for filtering red blood cells from a blood sample in order to obtain a plasma sample.

The cartridge 11 is also in dry form in the first reaction zone Z1 located below the filtration membrane 110 (on the other side of the membrane different from the capillary output 131), the above-mentioned reagents (one or more livers). Includes a first detection pad 111 that includes (suitable for optically detecting enzymes) (FIGS. 5 and 7). Here, the first detection pad 111 contains reagents particularly suitable for detecting AST. The cartridge 11 also includes a second detection pad 112 containing another liver enzyme, a reagent suitable for detecting ALT, in a second reaction zone Z2 located below the filtration membrane 110. The first detection pad 111 and the second detection pad 112 are arranged side by side below the filtration membrane 110, for example in the form of two separate reagent dots (FIG. 5).

Therefore, after passing through the filtration membrane 110, a part of the blood sample that is effectively formed into the plasma sample due to the filtration reaches the first detection pad 111 and permeates. Another portion of the filtered blood sample reaches and penetrates the second detection pad 112.

Upon contact with AST, the reagent contained in the first detection pad 111 produces a product (hydrogen peroxide in this case) that reacts with an indicator (here, MAOS Trender reagent) as a result of one or more chemical reactions. Produce and this indicator is colored if this product is present. Therefore, thanks to this reaction in the presence of AST or these reactions, this indicator is colored and allows liver enzyme detection.

The color change of the indicator is optically detected by a blood test reader 3 configured to measure the light reflectance, specifically the diffuse light reflectance, in the first reaction zone Z1. For this purpose, the blood test reader 3 includes a light source and a photodetector. Here, a light source, such as a light emitting diode, provides an emission spectrum suitable for detecting the color change of the indicator, even without spectral filtering of the light reflected by the first reaction zone Z1 of the blood test disposable 10. Have. For example, if the indicator in the colored form selectively absorbs yellow light, the light source may emit predominantly yellow light (ie, the emission spectrum of the light source is at least a portion of the absorption spectrum of the indicator in the colored form. May match).

The expression "light reflectance" is used to indicate any amount of light intensity or spectral intensity or power reflected by an object or surface as compared to the power or intensity or spectral intensity illuminated by the object or surface. You should be careful. In particular, the expression "light reflectance" may refer to the reflectance, reflectance, spectral reflectance, or incident-to-reflectance ratio of light whose spectrum is predominantly contained in a given limited wavelength range. Also, the light reflectance of reaction zone Z1 is measured or estimated by comparing the luminosity of the light reflected by this reaction zone with the luminosity of the light reflected by another reference zone (used as a "blank"). May be done.

The blood test reader 3 is calibrated and programmed to determine the AST concentration in a blood sample taken from a subject from the light reflectance measurements described above.

Similarly, the reagent contained in the second detection pad 112 upon contact with the ALT is a product (again, hydrogen peroxide) that reacts with the indicator (again, the MAOS Trender reagent) as a result of one or more chemical reactions. Hydrogen peroxide) is produced and this indicator is colored in the presence of this product.

The blood test reader 3 also measures the light reflectance in the second reaction zone Z2 (eg, in the same manner as the measurement of the light reflectance in the first reaction zone Z1), and from this measurement in the blood sample. It is configured to determine the ALT concentration.

Here, the types of reagents and the chemical reactions associated with the detection of this enzyme are shown in detail with reference to FIG.

The first detection pad 111 (for AST detection) contains L-aspartic acid and α-ketoglutaric acid. When a portion of the filtered blood sample containing AST comes into contact with this pad, L-aspartic acid and α-ketoglutaric acid react together, and AST acts as a catalyst (FIG. 9). This first reaction R1 produces oxaloacetate, which is then decomposed into pyruvic acid and carbon dioxide in reaction R2.

The first detection pad 111 further comprises a catalyst, namely pyruvate oxidase and peroxidase. The pyruvate produced by reaction R2 reacts with phosphate, oxygen and water contained in the filtered blood sample (a sample that may be mixed with a buffer) and acetyls during the reaction called R3. Produces phosphoric acid, carbon dioxide and hydrogen. Reaction R3 is catalyzed by pyruvate oxidase.

Here, the indicator contained in the first detection pad 111 is colored when oxidized. Therefore, in the presence of hydrogen peroxide produced by reaction R3, the indicator is oxidized and colored during reaction R4. Reaction R4 is catalyzed by peroxidase. The indicator may be, for example, the MAOS Trinder reagent (detailed formula above). In fact, this indicator has been shown to enable sensitive and reliable enzyme concentration measurements. In this case, the emission spectrum of the light source used to detect the MAOS state change may contain predominantly orange light (this spectrum extends predominantly around a wavelength of 610 nanometers) or may be filtered. Thereby, reflectance measurements are made, for example, in a wavelength range that extends around a wavelength of predominantly 610 nanometers, including between predominantly 550 nanometers and 650 nanometers.

Here, the first detection pad 111 comprises two separate sub-pads, an upper pad containing reagents (L-aspartic acid and α-ketoglutaric acid) and a lower pad containing catalysts (pyruvic acid oxidase and peroxidase). include. The lower pad is located below the upper pad. The filtered blood sample (ie, plasma sample) first reaches the upper reagent pad and then flows or moves to reach the lower catalyst and indicator pad when pyruvic acid is produced. The upper pad may be implemented in the form of a substrate that extends over the lower pad (covers a larger area than the area of the lower pad). However, in some embodiments, reagents, catalysts and indicators can also be mixed in a single dry detection pad rather than being separated from each other (for better storage), or as described above. It is also possible to disperse to different pads in a different way than the method.

The second detection pad 112 (for ALT detection) contains L-alanine (instead of L-aspartic acid) and α-ketoglutaric acid. It also contains the same catalysts and indicators as the first detection pad 111, namely pyruvate oxidase and peroxidase and MAOS Trinder reagent. Here, the second detection pad 112, like the first detection pad 111, includes an upper pad containing reagents (L-alanine and α-ketoglutaric acid), a catalyst (pyruvic acid oxidase and peroxidase), and an indicator (MAOS). ) Including the lower pad and including.

The reaction that occurs in the second detection pad 112 when a portion of the filtered blood sample penetrates the second detection pad 112 is as follows. First, L-alanine and α-ketoglutaric acid react together, and ALT acts as a catalyst (reaction R5). This reaction R5 produces pyruvate and glutamate. The pyruvate then reacts with the phosphate, oxygen and water contained in the filtered blood sample (which may be mixed with the buffer) and the reaction described above (with the detection of AST). Produces acetyl phosphate, carbon dioxide and hydrogen hydrogen during the same reaction as R3. The hydrogen peroxide produced by reaction R3 then oxidizes the indicator during the same reaction as reaction R4 described above, and the indicator is colored.

Optionally, the blood test disposable 10 is also placed between the filter membrane and the first reaction zone along a path followed by a filtered blood sample, as schematically shown in FIG. A hemolysis check optical port may be included (in FIG. 10 this optional element is identified by reference number 122). This optical port may be implemented in the form of a window with a passage for the filtered blood sample behind. To check if the blood sample is significantly hemolyzed, the blood test reader is therefore configured to measure light reflectance and / or transmittance in a wavelength range suitable for detecting hemoglobin. In some cases (here, the light reflectance at the hemolysis check optical port). For example, this light reflectance may be measured, for example, in a wavelength range that extends primarily around 410 nanometers, including between 390 and 430 nanometers. This light reflectance measurement may be realized by using a light emitting diode that emits blue light and using a photodetector such as a photodiode.

The blood test reader 3 uses the reflectance and / or transmittance measurements to check, for example, whether the light reflectance in the hemolytic check optical port exceeds a given threshold, so that the filtered blood sample has the above-mentioned wavelengths. It may be programmed to test for significant absorption of light in the range (indicating significant hemolysis). Also, when it exceeds the threshold, the blood test reader may issue an error message indicating that the liver enzyme concentration measurement has failed. This error message may be transmitted by the blood test reader, for example by making an audible beep, turning on a particular optical indicator, or displaying an error message on the display screen of the blood test reader. Thereby, the operator can directly recognize this failure. The blood test reader may also be configured to send the above error message to the control and processing system 4. In any case, when the blood test reader 3 determines that the filtered blood sample significantly absorbs light in the wavelength range described above, it prohibits the transmission of liver enzyme concentration measurement results (results of those measurements). Do not send or output). In fact, high absorption in the wavelength range described above indicates that the blood sample can significantly hemolyze and nearly change its color, thus causing liver enzyme measurement errors (liver enzymes are colored indicators). Because it is detected using). Thereby, thanks to the hemolysis check port of the blood test disposable 10 and the associated detection system of the blood test reader 3, the certainty of liver enzyme measurement is improved.

The blood test disposable 10 also includes on-board control 119 (FIGS. 5 and 8) of catalytic activity described above. This activity control is based on liver enzyme detection, i.e. the same colorimetric detection scheme for reactions R3 and R4 described above. To check the activity and integrity of the catalysts and indicators associated with their detection reactions, onboard control 119 comprises oxaloacetate predicted to be detected by those catalysts and indicators (see Reactions R2 through R4). ..

More specifically, the onboard control 119 is:
-A dry control substrate 116 containing oxaloacetate and
-Includes a dry control pad 117 containing the catalysts described above (ie, pyruvate oxidase and peroxidase) and an indicator (here MAOS).

Since the control substrate 116 and the control pad 117 are separate from each other, the oxaloacetate and the catalyst / indicator do not mix with each other (and therefore do not react with each other) during storage of the disposable 10. Here, the control pad 117 is arranged below the control board 116.

The cartridge 11 further includes a breakable blister 114 configured to break when the blood test disposable 10 is inserted into the blood test reader 3. The blister 114 is fluid-connected to the onboard control 119 here using a supply pipe 115. The supply tube 115 extends from the blister 114 to the output end of the tube located directly above the control board 116 (on the other side of the control board different from the control pad 117). When the blister 114 breaks due to the insertion of the disposable 10 into the reader 3, the buffer flows into the supply tube 115 to reach the onboard control, where it penetrates the control board 116 and then the control pad 117. The oxaloacetate of the control substrate 116 is mixed with the catalyst and indicator of the control pad 117. Then, unless the catalyst or indicator is defective, reactions R2 to R4 occur and the color of the indicator changes. This color change is optically detected by the blood test reader 3.

For more reliable and sensitive detection of this color change, the onboard control 119 is used to record a "blank", which is a reference value for light reflectance, without changing color in the absence of oxaloacetate. Includes pad 118. The light reflection characteristics of the reference pad 118 are close to those of the control pad 117, eg, within 20%, and even within 10% (at least in the visible range), when the color of the indicator has not changed yet. Or in part of the visible range). For example, the reference pad 118 may be identical to the control pad 117, except that it lacks the catalysts (pyruvate oxidase and peroxidase) and / or indicators described above.

The buffer (initially contained in Blister 114) is an aqueous solution containing phosphate. It may have a pH contained between 6 and 8, or even between 6.5 and 7.5. It may include TRIS, where the concentration of TRIS is between 0.05 mol / liter and 1 mol / liter, or even between 0.1 mol / liter and 0.5 mol / liter.

The cartridge 11 is configured such that if the blister breaks, the buffer not only flows through the control substrate 116, but also the buffer mixes with the plasma sample after reaching the detection area where the plasma sample mixes with the reagents and catalyst. May be done. This is beneficial because the above-mentioned types of buffers (TRIS buffers with the above-mentioned concentrations and pH) contribute to fast and accurate detection of AST and ALT when mixed with plasma samples. In particular, the cartridge is:
-The front of the cartridge, which includes the blister 114 and the onboard control 119,
-It may include detection pads 111 and 112, and a permeable membrane 123 that separates from the rear of the cartridge into which the capillaries 13 enter when the plug 12 is inserted into the cartridge 11.

FIG. 10 outlines the various steps involved in detecting liver enzymes and controlling the activity of catalysts and indicators as a flow chart.

The blood test disposable 10 and associated reader 3 described above are specifically configured to detect AST and ALT.

Similarly, in other embodiments (not shown), blood test disposables and readers are such to measure the chemical activity of only one (but not both) of those two enzymes, eg, AST alone. Can be configured in. Alternatively, they may be configured to measure the chemical activity of another liver enzyme such as GGT, or to measure the chemical activity of AST and GGT, or AST, ALT and GGT.

In some embodiments, the blood test disposable and reader can be configured to measure the number of platelets in a collected blood sample in addition to the chemical activity of AST (and ALT). To measure the platelet count, some of the blood sample taken by the capillary may remain unfiltered and longer cartridges are used with the capillary output and filter membrane for the platelet measurement element. It is also possible to leave a space between them.

In some embodiments, in addition to the blood test reader 3 and associated disposable 10 (configured to determine the AST / ALT concentration in the subject's blood), the system was also taken from the subject. It may include another clinical site immediate device configured to determine platelet concentration in a blood sample.

FIG. 11 shows, as a flow chart, a first embodiment of a method for determining parameters related to a subject's liver health. FIG. 12 simplifies the same method as in FIG. 11, but shows some more concrete and exemplary.

In this method, the control and processing system 4 controls the elastography device 2 and the blood test reader 3 so that the mechanical parameters are measured first (steps S10 to S1). Blood samples are then taken from the subject and liver enzyme concentrations are measured (steps S20-S2) or not, depending on the values of the mechanical parameters so measured. In this first embodiment, blood tests are performed only when it is expected to improve the determination of parameters related to the health of the subject's liver.

In FIG. 11, various steps of this method are shown for time t. In this figure, the direction in which the time t elapses corresponds to the vertical downward direction. The step in question is aligned perpendicular to the entity performing the step being considered. For example, step S1 is performed by the control and processing system 4, while step S100 is performed by the operator 100.

The method starts from step S10. In step S10, the control and processing system 4 controls the elastography device 2, in particular the elastography module 21, so as to initiate the elastography measurement process.

In response, in step S11, the elastography module 21 displaces a portion of the subject's body on which the probe 22 is placed to guide the operator 100 and place it on the probe 22 in front of the subject's liver. Generate signals suitable for exploration (especially ultrasonic signals). The operator interface 5 provides guidance information thus obtained in step S12, such as A-mode ultrasound images and other information useful for elastographic measurements (eg, contact force acting on the tip of probe 22). Here, by displaying it, the operator 100 is informed. In step S12, the operator interface 5 may prompt the operator to make an elastographic measurement.

Then, in step S100, the operator 100 triggers an elastography measurement, here a transient elastography measurement. More precisely, he has low-frequency transient elastic waves (eg, shear waves) and ultrasonic shots sent to track how the subject's tissue is moved by the elastic waves. Trigger radiation. In step S13, the elastography module 21 acquires and processes the received signal in response to the signal, and determines the value of the above-mentioned mechanical parameter (for example, liver hardness). This transient elastography measurement can be repeated several times. Further, in step S13, another ultrasonic echo signal obtained (based on the ultrasonic echo signal obtained during the transient elastography measurement and / or, for example, in the interval between two consecutive transient elastography measurements). The CAP value is determined (by either).

The values of the mechanical parameters and the CAP values so measured are then sent to the control and processing system 4, which acquires them in step S1.

Then, in step _ST , the control and processing system 4 tests whether the value of the mechanical parameter meets a given criterion.

Here, in the control and processing system 4, the values of the mechanical parameters (for example, liver hardness) acquired in step S1 may be a value in which the liver does not suffer from a health disorder on average and a value in which the liver suffers from a health disorder. When the threshold corresponding to the boundary between a possible value is exceeded, it is determined that the value does not meet the criteria.

When the mechanical parameter is Young's modulus E, this threshold value may be in the range of, for example, 6-7 kPa. In fact, a Young's modulus of less than 6-7 kPa indicates that the subject under test does not appear to suffer from liver fibrosis, regardless of the concentration of liver enzymes in the subject's blood.

When the above criteria are met, the control and processing system 4 executes S3'. In step S3', the control and processing system 4 takes into account the value of at least one mechanical parameter, regardless of the concentration of liver enzymes (ie, AST, ALT or GGT) in the subject's blood, in the subject's liver. Determine the parameters related to your health. Also, when this criterion is met, no blood sample is taken from the subject and is not analyzed.

Conversely, if the criteria described above are not met, after step _ST , in step S20, the control and processing system 4 initiates the liver enzyme measurement process. In particular, in step S20, the control and processing system 4 controls the operator interface 5 to collect information and / or a capillary blood sample from the subject indicating that liver enzyme concentration measurements are recommended and this blood. It conveys information prompting the operator 100 to start analyzing the sample.

Then, in step S200, the operator 100 collects a capillary blood sample from the subject by directly using the capillary tube 13 fixed to the blood test disposable 10. After that, the operator introduces the blood test disposable 10 filled with the blood sample into the blood test reader 3.

In step S23, the blood test reader 3 determines the concentration of at least one liver enzyme in this blood sample. Here, the blood test reader 3 specifically determines the concentrations of AST and ALT using the above-mentioned colorimetric detection method carried out in the blood test disposable 10.

The blood test disposable 3 then sends one or more values of the liver enzyme concentration measured in step S23 to the control and processing system 4, which or the control and processing system 4 obtains it or them in step S2.

Then, in step S3, the control and processing system 4 takes into account at least one of the mechanical parameter values obtained in step S1 and the liver enzyme concentration value obtained in step S2. Determine parameters related to the subject's health. The control and processing system 4 then outputs data representing the parameters thus determined. These data may be transmitted to the operator 100 by, for example, the operator interface 5. They may also be sent to a personal electronic health card (for storing the above data in this microcircuit card), or a storage device or system such as a non-local / distributed health data storage system.

In the method of FIG. 11, the operation of the elastography device 2 and the operation of the blood test device 3 are clearly coordinated with each other (because, depending on the mechanical parameter value, the liver enzyme measurement is started after the mechanical parameter measurement. ). Also, the operation of the elastography device 2 and the operation of the blood test device 3 are simultaneous, that is, they occur during a given limited time frame.

More specifically, in the method of FIG. 11, the startup time lag _TS between the start of step S10 and the start of step S20 is limited (due to the way the control and processing system is programmed). Is clear. In practice, a major part of this duration is the operator performing the necessary operations (probe positioning, measurement triggering, possible iterations of elastographic measurements) to explore the subject's liver hardness. Corresponds to measurement step S100. In practice, _TS is usually between 2 and 5 minutes and is generally less than 10 minutes (or at least less than 20 minutes).

Further, in the method of FIG. 11, the time lag Tm between the measurement of the mechanical parameter (at the end of step S100) and the measurement of the liver enzyme concentration (at the end of step S23) is also limited. In fact, the main part of this duration is:
-Operations performed by the operator 100 (capillary blood sampling, insertion of the blood test disposable 10 into the blood test reader, which usually takes 1-3 minutes).
-Here corresponds to the time required to complete the chemical reaction detection, which is usually less than 5 minutes (or at least less than 10 minutes), thanks to the particular characteristics of the blood test disposable 10 described above.

Thus, in practice, Tm is typically between 6 and 15 minutes (and generally less than 30 minutes).

Therefore, the total time _TT required to perform the entire method of FIG. 11 is typically between 7 and 30 minutes, often between 10 and 20 minutes (in any case). Also less than an hour).

In the following, the determination of the parameters related to the health condition made in step S3 is shown in detail.

Whether the health-related parameters determined in step S3 are considered to be healthy in the subject's liver, or conversely suffer from a given disease or inflammation such as fibrosis or steatosis. It may be in the form of information that clearly indicates.

In addition, the health state-related parameters determined in step S3 may be more stepwise clarified whether or not the subject's liver is considered to be healthy, for example, in the form of a disease stage. For example, this disease stage is the widely used fibrosis stages F0 to F4 (F0: no fibrosis; F1: minimal fibrosis; F2: moderate fibrosis; F3: severe fibrosis; F4 :. It can also be the most severe fibrosis / cirrhosis).

The health-related parameters determined in step S3 may be related to fibrosis, but steatosis (CAP values have been found to be very useful) and / or hepatitis (liver enzymes in this regard). The value of the concentration of may be related to (providing very useful information).

In step S3, the parameters related to the liver health of the subject are:
-Calculate the value of the benchmark parameter (score) depending on at least the mechanical parameter and the concentration of liver enzyme described above, and then-the value of the benchmark parameter thus obtained-one or more thresholds (eg, for example). Negative predictive value, and positive predictive value)
-It may be determined by determining the parameters related to the liver health of the subject from the results of one or more comparisons in question.

Also, in some embodiments, different disease stages can be associated with different value ranges of the benchmark parameters described above, respectively.

Also, this health-related parameter determined in step S3 depends on at least the mechanical parameters and the concentration of liver enzymes described above, and is the value of the benchmark parameter representing the fact that the subject's liver is to some extent healthy. It can also take a form.

In other words, the health-related parameters do not correspond to the final diagnosis itself, but may also be intermediate benchmark parameter values useful for the medical professional to make a diagnosis of the subject's liver.

The benchmark parameters described above can also be, for example, the "FAST" parameters described above calculated according to equation F1. As mentioned above, this benchmark parameter depends on the liver hardness LSM (ie, Young's modulus E), CAP, and the concentration of AST in the subject's blood.

Benchmark parameters, or more generally parameters related to the subject's liver health, can be determined taking into account the concentration of ALT in the subject's blood. Therefore, the benchmark parameters in question may depend on the liver hardness LSM, optionally CAP, the concentration of AST in the subject's blood, and the concentration of ALT in the subject's blood.

Other characteristics of the subject's blood, such as platelet concentration, may also be taken into account when determining parameters related to the subject's liver health.

Other clinical parameters relating to the subject, such as age and gender, may also be taken into account when calculating the benchmark parameters (scores) described above (score formulas may include such additional clinical parameters). ..

For example, parameters related to the health of the subject's liver can be determined taking into account the concentration of AST in the subject's blood, the concentration of ALT and the concentration of platelets, the age of the subject, and the liver hardness LSM.

To this end, in one embodiment, the control and processing system 4 is:
-The values of the "FIB-4" benchmark parameters can be calculated and then-the parameters related to the subject's health can be determined based on the FIB-4 values and the LSM values obtained in step S1.

The FIB-4 benchmark parameters are defined in the abstract of the following paper: "Development of a simple noninactive index to predict fibrosis patients with HIV / HCV co-inject". K. Etc., Hepatology 2006, vol. 43, pp. 1317-1325.

In such embodiments, the FIB-4 value may be used, for example, to confirm or, conversely, invalidate the health condition initially identified based on the LSM value.

The FIB-4 value may also be used to improve prior health identification based on the LSM value, in particular in the LSM "gray range" where the LSM value is between the negative predictive value and the positive predictive value. When included, it helps to distinguish between health conditions.

The FIB-4 value may also be taken into account by calculating the overall score according to both the FIB-4 value and the LSM value.

In step S3, various benchmark parameters (various scores) can be used to determine parameters related to the health of the subject, as shown in FIG. This is useful because given benchmark parameters may be more appropriate than others in order to infer reasonable information related to the subject's liver health, depending on the health status identified. be.

For example, if benchmark parameters based on logistic regression are used, each of those benchmark parameters will be able to make a binary decision. For example, it may be possible to determine whether the subject's liver suffers from F4 fibrosis with the first benchmark parameter, while the subject's fibrosis stage is F0-F1 or higher. It can be determined by benchmark parameters. Subject by using benchmark parameters selected from a variety of benchmark parameters, depending on the health condition identified and / or such prior characterization of the subject's liver condition. More detailed or more complete determination of parameters related to liver health can be achieved. Since the subject's health or physical condition is not known in advance, the selection of more appropriate benchmark parameters is the value of the mechanical parameter (eg liver hardness) and / or the value of the ultrasonic attenuation parameter obtained in step S1. It is possible to base on the results of the preliminary stage consisting of.

For this purpose, in some embodiments:
-In the memory of the control and processing system 4, various formulas are associated with different ranges of values of at least one mechanical parameter, and each formula corresponds to a given benchmark parameter for liver health assessment. Each formula allows the corresponding benchmark parameters to be calculated from at least one mechanical parameter and from the concentration of at least one liver enzyme in the blood sample.
-The control and processing system 4 compares the values of at least one mechanical parameter previously measured with respect to the subject's liver to the range of values associated with each of these various benchmark parameters, thereby the above benchmark parameters. Programmed to select one of them,
-The control and processing system 4 is programmed in step S3 to calculate the value of the previously selected benchmark parameter according to the equation associated with this benchmark parameter.

This method of determining the parameters related to the health condition of the subject is shown as an example in FIG.

In this example, when the Young's modulus E of the subject's liver is less than a given threshold equal to 6.1 kPa here, a first benchmark parameter called score a is selected. Then, the value of the score a is calculated in consideration of the value of E and the value of the concentration of AST and ALT in the blood of the subject. When the value of the score a is less than the first threshold t1, the control and processing system 4 determines that the fibrosis stage of the subject's liver is F0 or F1. Also, when the value of score a is above the second threshold t2, the control and processing system 4 determines that the fibrosis stage of the subject's liver is F2, F3 or F4 (included between F2 and F4). decide. When the value of the score a is between t1 and t2 (“gray zone”), the control and processing system 4 outputs data indicating that the parameters related to the health condition of the subject's liver are not decidable.

Conversely, when the Young's modulus E of the subject's liver is above the threshold above equal to 6.1 kPa, a second benchmark parameter called score b is selected. The score b is then calculated taking into account the value of E and the values of AST and ALT concentrations in the subject's blood. When the value of the score b is less than the third threshold t3, the control and processing system 4 determines that the fibrosis stage of the subject's liver is F0 or F1. Also, when the value of the score b is above the fourth threshold t4, the control and processing system 4 determines that the fibrosis stage of the subject's liver is F2, F3 or F4. Also, when the value of score b is between t3 and t4 (“gray zone”), the control and processing system 4 outputs data demonstrating that parameters related to the health of the subject's liver are not determinable. do.

FIG. 14 shows a computational and inspection sequence performed in a particular example of the realization of the method of FIG. In this example, the method for determining the parameters related to the subject's liver health is to determine whether the subject's liver fibrosis stage is F4 or less (F0 to F3). Especially intended. Therefore, the threshold of E involved in step _ST is relatively high, for example equal to 10.9 kPa.

When E is less than 10.9 kPa, after step _ST , the control and processing system 4 indicates that the subject's liver fibrosis stage is included between F0 and F3 (equivalent to F0, F1, F2 or F3). , Perform step S3'to determine directly, without further calculation (in the case of this particular example).

Conversely, if E is above 10.9 kPa, after step _ST , the control and processing system 4 initiates liver enzyme measurements (steps S20-S2). Also, in step S3, it then calculates the score value taking into account the values of E and the concentrations of AST and ALT. When the score value is less than the threshold t1', the control and processing system 4 determines that the subject's liver fibrosis stage is included between F0 and F3. When the score value is above the threshold t2', the control and processing system 4 determines that the subject's liver fibrosis stage is F4. Otherwise, the control and processing system 4 outputs data indicating that the parameters related to the health condition of the subject's liver are not decidable (“gray zone”).

FIG. 15 schematically illustrates a second embodiment of a method for determining parameters related to a subject's liver health according to the techniques of the present disclosure.

In this figure, the method is shown in the form of a flow chart having the same rules as in FIG. 11 (particularly the direction in which the time t elapses corresponds to the vertical downward direction).

In the second embodiment, the liver enzyme measurement process and the mechanical parameter measurement process are started in substantially parallel with a short delay _TS'between their respective initiations. The liver enzyme measurement process is first initiated (in step S20'). Then, when a blood sample is taken from the subject (in step S200) and when the blood test disposable 10 is introduced into the reader 3 (at the beginning of step S23), a mechanical parameter measurement process (in step S10). Can be started without delay.

Therefore, the time required for the detection chemical reaction to occur, usually between 5 and 10 minutes, is used to measure the mechanical parameters described above in parallel. This makes it possible to reduce the total time _TT'required to determine parameters related to the subject's liver health.

Therefore, the method of FIG. 15 begins in step S20'during which the control and processing system 4 initiates the liver enzyme measurement process. In particular, in step S20', the control and processing system 4 is directed to send information prompting the operator 100 to take a capillary blood sample from the subject and start analyzing the blood sample. Control the interface 5. In step S22, by displaying here, the operator interface 5 conveys the above information to the operator 100. Then, in step S200, the operator collects a capillary blood sample from the subject and introduces the blood test disposable 10 filled with the blood sample into the blood test reader 3 (similar to the method of FIG. 11). Then, step S23 for analyzing the blood sample is started. At the start of step S23 immediately after the blood test disposable 10 is inserted into the blood test reader 3, the blood test reader 3 is inserted into the control and processing system 4, and the blood test disposable 10 is inserted into the blood test reader 3. Send information that clearly indicates that. In step S24, this information is received by the control and processing system 4. Upon receiving this information confirming that the blood sample analysis has started, the control and processing system 4 initiates the mechanical parameter measurement process in step S10. Here, this measurement process includes steps S10, S11, S12, S100, S13, and subsequent S1 already described above with reference to FIG.

During the execution of steps S10 to S1, the chemical analysis of the blood sample continues, and at the end of step S23, it finally leads to the determination of the respective values of the AST concentration and the ALT concentration in the blood of the subject. These values are then transmitted to the control and processing system 4 which will acquire them in subsequent step S2. Then, step S3 is executed as described above with reference to FIG.

In the method of FIG. 12, the time _TS'is usually between 1 minute and 5 minutes. Also, at the beginning of the liver enzyme measurement (ie, step S200) and at the end of all serological or mechanical measurements (here, depending on the latest, at the end of step S23 or in step S13). The total time Tm'with (corresponding at the end) is usually in the range of 4 to 10 minutes. The total time _TT'required to perform this method is shorter than the total time _TT required to perform the method of FIG. 11, typically between 5 and 20 minutes, 7 minutes. Often between and 15 minutes (less than an hour anyway).

1, 1', 1 "system 2, 2', 2" elastography device 3 blood test reader 4, 4', 4 "control and processing system 5, 5'operator interface 6, 6'casing 7 computational resource 8 Remote device 9 Connection 10 Blood test Disposable 11 Cartridge 12 Plug 13 Capillary 21 Elastography module 22 Probe 41 First part 42 Second part 100 Operator 110 Filter membrane 111 First detection pad 112 Second detection pad 114 Blister 115 Supply tube 116 Control board 117 Control pad 118 Reference pad 119 Onboard control 121 Cartridge end face 123 Permeable film

Claims (21)

A system (1; 1'; 1 ") for determining parameters related to the health condition of the subject's liver.
-With an elastography device (2; 2'; 2 ") configured to measure at least one mechanical parameter associated with shear wave propagation in the liver.
-A blood test reader (3) and a blood test disposable (10) associated with a blood test reader (3), the blood test disposable receives a capillary blood sample and puts it in the blood test reader (3). A blood test reader and a blood test disposable-at least a processor and a memory are configured to be inserted and the blood test reader is configured to determine the concentration of at least one liver enzyme in the blood sample. Including control and processing system (4; 4'; 4 ") including
-Blood test disposable (10) is
-A capillary tube (13) for collecting the blood sample from the finger, and
-Contains reagents suitable for detecting the at least one liver enzyme in the blood sample.
-The blood test reader (3) is operably connected to the elastography device (2; 2'; 2 ").
The control and processing system has the following steps,
-S1-Obtaining the value of the at least one mechanical parameter measured by an elastography device (2; 2'; 2 "),
-S2- Obtaining the value of the concentration of the at least one liver enzyme in the blood sample collected from the subject as measured by the blood test reader (3).
-S3- Taking into account both the value of the at least one mechanical parameter and the value of the concentration of the at least one liver enzyme, the parameters related to the health condition of the subject's liver are determined, and the value of the subject's liver is determined. It is programmed to output and perform data representing the parameters related to the health condition.
The parameter related to health status is the health stage or illness stage identified among the various stages (F1, F2, F3, F4) of a given health status classification, or at least one mechanically said. It is a benchmark parameter that takes into account both the value of the parameter and the value of the concentration of at least one liver enzyme, or is represented by both the health stage or disease stage and the value of the benchmark parameter.
The control and processing system (4; 4'; 4 ") allows the measurement of the at least one mechanical parameter and the measurement of the value of the concentration to be performed according to a predefined time sequence or with a predefined condition. A system (1; 1'; 1 ") configured to control an elastography device (2; 2';2") and a blood test reader (3) as performed according to the measurement procedure. The control and processing system (4; 4'; 4 ") has an elastography device (2; 2'; 2") and a blood test reader (3), respectively, within a single time frame. It is programmed to control the elastography device (2; 2'; 2 ") and the blood test reader (3) to initiate the process and the blood test process, including the collection and analysis of the blood sample. , The system according to claim 1 (1; 1'; 1 "). The system according to claim 2, wherein the control and processing system (4; 4'; 4 ") is programmed so that the time frame has a duration of up to 30 minutes ( _TS ; _TS '). 1; 1'; 1 "). The invention according to any one of claims 1 to 3, wherein the blood test disposable (10) is configured such that the blood sample collected using the capillary tube (13) has a maximum amount of 60 microliters. System (1; 1'; 1 "). The control and processing system (4; 4'; 4 ") is performed in the following steps,
-S10-Controlling the elastography device so that the elastography device (2; 2'; 2 ") starts the elastography measurement process, and then-the above-mentioned step S1 and then-the above-mentioned acquired in step S1. If the value of at least one mechanical parameter meets a given criterion
-S3'-Regardless of the concentration of the at least one liver enzyme in the subject's blood, taking into account the values of the at least one mechanical parameter, determine the parameters related to the subject's liver health. Outputting data representing the above parameters, while
-If the value of at least one mechanical parameter does not meet the criteria
-S20-Information clearly stating that a blood test is recommended for the subject's liver health assessment, and / or collecting a capillary blood sample from the subject for the operator (100) and analyzing this blood sample. Controlling the operator interface (5; 5') so that the operator interface (5; 5') sends information prompting it to start, and then-the step S2 and then-the step S3. Programmed to run,
The system according to any one of claims 1 to 4 (1; 1'; 1 "). The control and processing system (4; 4'; 4 ") addresses the boundary between values on average that the liver does not have health problems and values that the liver may have health problems. The system according to claim 5, wherein when the value of the at least one mechanical parameter exceeds the threshold value, it is programmed to determine that the value of the at least one mechanical parameter does not meet the criteria. 1; 1'; 1 "). The control and processing system (4; 4'; 4 ") is performed in the following steps,
-S20'-Once a capillary blood sample is taken from the subject and then the blood test disposable (10) is inserted into the blood test reader (3), the operator (100) can start the analysis of this blood sample. To control the operator interface (5; 5') so that the operator interface (5; 5') transmits the information prompting the operator.
-S10-Controlling the elastography device (2; 2'; 2 ") so that the elastography device (2; 2';2") initiates the elastography measurement process, and then-steps S1 and S2. And then
-Programmed to perform step S3 and above.
The system according to any one of claims 1 to 4 (1; 1'; 1 "). -In the memory of the control and processing system (4; 4'; 4 "), various formulas are associated with different ranges of the values of the at least one mechanical parameter, and each formula is for liver health assessment. Corresponding to a given benchmark parameter, each formula allows the corresponding benchmark parameter to be calculated from the at least one mechanical parameter and the concentration of the at least one hepatic enzyme in the blood sample.
-The control and processing system (4; 4'; 4 ") sets the values of the at least one mechanical parameter previously measured with respect to the subject's liver to the range of values associated with their various benchmark parameters, respectively. Programmed to select one of those benchmark parameters by comparison,
-The control and processing system (4; 4'; 4 ") is programmed in step S3 to calculate the value of the previously selected benchmark parameter according to the equation associated with this benchmark parameter.
The system according to any one of claims 1 to 7 (1; 1'; 1 "). The system (1; 1'; 1 ") according to any one of claims 1 to 8, wherein the capillaries (13) are fixed to a part (12) of the disposable (10). Blood test disposable (10),
-Cartridge (11) containing the reagent and
-Including removable plug (12)
Capillaries (13) are secured to removable plugs (12) or cartridges, and the plugs (12) and cartridges (11)
-So that the plug (12) can be removed from the cartridge (11) and then reinserted into the cartridge, and-when the plug (12) is inserted into the cartridge (11), the capillaries The system (1; 1'; 1 ") according to claim 9, wherein (13) is housed inside a disposable (10) and is configured such that a blood sample collected by a capillary is in contact with the reagent. The at least one liver enzyme is one of aspartate aminotransferase (hereinafter "AST"), alanine aminotransferase (hereinafter "ALT"), and γ-glutamyl transferase (hereinafter "GGT").
The reagent is suitable for optically detecting the at least one liver enzyme.
A blood test disposable (10) is configured such that at least a portion of the blood sample mixes with the reagent in the first reaction zone (Z1).
A blood test reader (3) uses light reflectance and / or transmittance measurements in a first reaction zone (Z1) to determine the concentration of said at least one liver enzyme in the blood sample. , At least including a light source and an optical sensor,
The system according to any one of claims 1 to 10 (1; 1'; 1 "). The blood test disposable (10) comprises a filtration membrane (110) configured to filter red blood cells from a blood sample taken from a subject to obtain a plasma sample, at least a portion of the plasma sample with said reagent. The system according to any one of claims 1 to 11 (1; 1'; 1 ") configured to be in contact. -The at least one liver enzyme is AST or ALT,
-The reagents contain α-ketoglutaric acid and L-aspartic acid or L-alanine, respectively.
-The blood test disposable (10) contains the following catalysts, pyruvate oxidase and peroxidase, and is colored when the at least one liver enzyme reacts with the product of a set of reactions that occur when mixed with the reagent. Including the indicator,
The system according to any one of claims 1 to 12 (1; 1'; 1 "). -The blood test disposable (10) comprises an onboard control (119) of the activity of the catalyst, the onboard control (119).
-A dry control substrate (116) containing oxaloacetate, and
-Includes a drying control pad (117) containing the catalyst and the indicator, which is separate from the control substrate (116).
-When the blood test disposable (10) is inserted into the blood test reader (3), or when the blood sample is received in the blood test disposable (10), the liquid is controlled substrate (116) and control pad. The blood test disposable (10) is configured to penetrate (117).
The system according to claim 13 (1; 1'; 1 "). 14. The liquid according to claim 14, wherein the liquid is a buffer contained in a destructible blister (114) configured to be destructible when the blood test disposable (10) is inserted into the blood test reader (3). System (1; 1'; 1 "). Further configured to determine the number of platelets in the blood sample or another blood sample taken from the subject, the control and processing system (4; 4'; 4 ") also takes into account the platelet count. , The system according to any one of claims 1 to 15 (1; 1'; 1 "), programmed to determine said parameters relating to the health of the subject's liver in step S3. A method for determining parameters related to a subject's liver health using the system (1; 1'; 1 "), wherein the system (1; 1';1") is used.
-With an elastography device (2; 2'; 2 ") configured to measure at least one mechanical parameter associated with shear wave propagation in the liver.
-A blood test reader (3) and a blood test disposable (10) associated with a blood test reader (3), the blood test disposable receives a capillary blood sample and feeds it to the blood test reader (3). A blood test reader (3) and a blood test disposable (10), configured to be inserted and configured to determine the concentration of at least one hepatic enzyme in the blood sample.
-Includes a control and processing system (4; 4'; 4 ") that includes at least a processor and memory.
-Blood test disposable (10) is
-A capillary tube (13) for collecting the blood sample from the finger, and
-Contains reagents suitable for detecting the at least one liver enzyme in the blood sample.
-The blood test reader (3) is operably connected to the elastography device (2; 2'; 2 ").
The method is the following steps,
-S100-For the liver of the subject to be tested, the value of the mechanical parameter was measured using an elastography device (2; 2'; 2 ").
-S1- Acquiring the value of the at least one mechanical parameter measured by the elastography device by the control and processing system (4; 4'; 4 "), and
-S200- Capillary blood samples are collected from the subject using the capillary tube (13), and then the blood test disposable (10) is introduced into the blood test reader (3).
-S2-A value of the concentration of at least one liver enzyme in a blood sample taken from a subject as measured by a blood test reader (3) by a control and processing system (4; 4'; 4 "). To get and
-S3-The control and processing system determines the parameters related to the liver health of the subject, taking into account both the value of the at least one mechanical parameter and the value of the concentration of the at least one liver enzyme. And including outputting data representing the above parameters related to the health condition of the subject's liver.
The parameter may be a health stage or disease stage identified among the various stages of a given health status classification (F1, F2, F3, F4), or the value of the at least one mechanical parameter and said. A benchmark parameter that takes into account both the value of the concentration of at least one liver enzyme, or is represented by both the health or disease stage and the value of the benchmark parameter.
The control and processing system (4; 4'; 4 ") allows the measurement of the at least one mechanical parameter and the measurement of the value of the concentration to be performed according to a predefined time sequence or with a predefined condition. A method of controlling an elastography device (2; 2'; 2 ") and a blood test reader (3) so that it is performed according to a measurement procedure. 17. The method of claim 17, wherein steps S100 and S200 are performed within a single time frame having the maximum preset duration (Tm; Tm'). 17. The method of claim 17, wherein the control and processing system outputs an error message when steps S100 and S200 are not performed within a single time frame having the maximum preset duration (Tm; Tm'). Steps S100 and S1 are executed first, and then the following steps,
-If the value of at least one mechanical parameter obtained in step S1 meets a given criterion.
-S3'-Control and processing system (4; 4'; 4 ") takes into account the value of the at least one mechanical parameter regardless of the concentration of the at least one liver enzyme in the subject's blood. , To determine the parameters related to the health condition of the subject's liver and output the data representing the said parameters related to the health condition, on the other hand.
-If the value of at least one mechanical parameter does not meet the criteria
-S20-The control and processing system (4; 4'; 4 ") clearly indicates that liver enzyme concentration measurement is recommended for the subject's liver health assessment, and / or to the operator (100). The operator interface (5; 5') is controlled so that the operator interface (5; 5') sends information prompting the subject to take a capillary blood sample and start the analysis of the blood sample. The method of any one of claims 17-19, comprising: The control and processing system (4; 4'4 ") may have values of the at least one mechanical parameter, on average, values that the liver does not suffer from health problems and the liver suffers from health problems. 20. The method of claim 20, wherein when the threshold corresponding to the boundary between the values is exceeded, it is determined that the value of the at least one mechanical parameter does not meet the criteria.

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