Method, device and computer readable medium for identifying hooking effect in immunonephelometry

Abstract

A method, device and computer readable medium for identifying hook effect in immunoturbidimetry. The method generates a reaction curve by measuring optical signals of an immune reaction of a to-be-measured substance in a sample within a predetermined time period, and determines whether the sample has hook effect by using distribution information of the reaction curve within the predetermined time period. The method only uses measurement information of a small predetermined time period in the entire reaction time to identify samples with hook effect, and terminates the detection in time, thereby accelerating the speed of immunoturbidimetry detection of samples with hook effect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technology of sample detection by immunoturbidimetry, and in particular to a method, device and computer readable medium for identifying the case of hook effect caused by too high concentration of the protein to be detected in the sample. BACKGROUND

[0002] In biochemical immunoassay, the ratio of antigen and antibody is often not appropriate, resulting in low detection results, or even false negative. Taking the case of excessive antigen as an example, when the amount of antibody-antigen precipitation is plotted against the amount of antigen, a hook-shaped (HOOK) curve (A) deviating from the normal curve (B) appears at the end of increasing antigen amount, which is called hook effect (see Figure 1 Hook effect is caused by excessive amount of antigen or antibody to be detected, resulting in soluble complex and deviation of detection results.

[0003] Turbidimetric inhibition immunoassay is a dynamic determination method of antigen and antibody binding. Turbidimetric inhibition immunoassay is divided into immunotransmission turbidimetry and immunoscattering turbidimetry. Antibody-antigen complex has scattering and shielding effect on light. Therefore, the amount of antibody-antigen complex is proportional to the change value of the intensity of transmitted light or scattered light. When the amount of antibody is constant, the change of light intensity is proportional to the content of antigen. Under certain conditions, the content of antigen in the sample can be obtained by detecting the intensity change of transmitted light or scattered light.

[0004] When the concentration of antibody is fixed, the amount of immune complex increases with the increase of the amount of antigen in the sample, and the turbidity of the reaction solution also increases. However, when the amount of the substance to be detected (i.e., antigen) in the sample is too large, hook effect occurs, i.e., the detection result is low, or even false negative.

[0005] At present, immunoturbidimetry is widely used in automatic biochemical analyzers. In order to obtain accurate detection results, it is necessary to identify the case of too high concentration of the substance to be detected in the sample to cause hook effect, and to detect the sample with hook effect again after appropriate dilution to obtain correct detection results. For example, Chinese patent application CN105339794A discloses a method of detecting and analyzing the light signal after complete reaction time by using two specific wavelengths, and judging whether the sample has hook effect by calculating the reaction rate. This method needs to detect at the same time by using two wavelengths, and can only make a judgment after the sample is completely detected.

[0006] Blood biochemistry tests, such as those for C-reactive protein (CRP) and serum amyloid (SAA), are widely used in clinical testing. Especially during seasons with a high incidence of respiratory diseases, CRP testing is often performed alongside complete blood counts (CBCs) to facilitate accurate clinical diagnosis. However, CRP testing typically takes longer than CBCs, resulting in longer waiting times for patients. If a sample exhibits a hook effect, retesting is required, further extending the waiting time. Therefore, there is a need for rapid identification of hook effects in samples to obtain accurate results quickly. Summary of the Invention

[0007] The purpose of this invention is to provide a method for identifying hook effects in immunoturbidimetry, so as to obtain judgment results more quickly and shorten the detection time of samples with hook effects.

[0008] Therefore, according to a first aspect of the present invention, a method for identifying the hook effect in immunoturbidimetry is provided. The method includes:

[0009] a) After the sample and reaction reagents are mixed to initiate the reaction of the analyte in the sample, the time intervals from T1 to T2 after the reaction begins are obtained. n T within the scheduled time period i Measurement value A at time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n To generate the reaction curve for that time period;

[0010] b) Estimate the concentration C of the analyte based on the reaction curve during this time period. e ;

[0011] c) Based on the estimated concentration C of the analyte e Obtain a pre-stored reference curve for this concentration;

[0012] d) Compare the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period to determine whether the sample has a hook effect.

[0013] In step a) of the method, T n The time T when the reaction has proceeded to 90% of its total reaction time is less than or equal to the time T. 90 Preferably, the time T is less than or equal to 70% of the total reaction time. 70 More preferably, the time T is less than or equal to 50% of the total reaction time. 50 .

[0014] or in step a) of the method, T n the corresponding time point at which the slope trend of the reference curve changes.

[0015] According to one embodiment, the predetermined time period is 10-70%, preferably 15-50%, more preferably 15-40% of the entire reaction time.

[0016] According to one specific embodiment, the time period for generating the analyte reaction curve is the time period from the start of the reaction to the time when the reaction has proceeded 10-70%, preferably 10-50%, more preferably 10-40%. That is, the analyte reaction curve is generated using the measurement values detected during the first 10-70%, preferably the first 10-50%, more preferably the first 10-40% of the reaction.

[0017] The distribution information in the method can be at least one selected from the group consisting of a measurement value feature, a reaction speed feature and a reaction acceleration feature.

[0018] According to the present application, if the distribution information of the reaction curve and the distribution information of the reference curve differ according to a predetermined feature, it is identified that the sample has a hook effect, and the detection is terminated.

[0019] According to one embodiment, the step of comparing the distribution information of the reaction curve in the predetermined time period with the distribution information of the reference curve in the corresponding time period comprises comparing the measurement values of the reaction curve in the predetermined time period with the measurement values of the reference curve in the corresponding time period.

[0020] According to another embodiment, the step of comparing the distribution information of the reaction curve in the predetermined time period with the distribution information of the reference curve in the corresponding time period comprises first-order differentiating the reaction curve and the reference curve to obtain a reaction speed curve and a reference speed curve, respectively, and comparing the reaction speed curve with the reference speed curve.

[0021] According to still another embodiment, the step of comparing the distribution information of the reaction curve in the predetermined time period with the distribution information of the reference curve in the corresponding time period comprises second-order differentiating the reaction curve and the reference curve to obtain a reaction acceleration curve and a reference acceleration curve, respectively, and comparing the reaction acceleration curve with the reference acceleration curve.

[0022] According to one embodiment, the step of comparing the distribution information of the reaction curve in the predetermined time period with the distribution information of the reference curve in the corresponding time period comprises comparing the values of the corresponding distribution information of the reaction curve and the reference curve at the same reaction time point, and / or comparing the average values of the plurality of corresponding distribution information of the reaction curve and the reference curve in the same time period.

[0023] According to one specific embodiment, in the above step d), the reaction curve is compared with the reference curve at T iMeasurement value A at time i Reference value A at the corresponding moment of the reference curve i(f) The response difference δ is calculated using the following formula. i %:

[0024] δ i % = [(A i -A i(f) ) / A i(f) ]×100%

[0025] When δ i When the percentage is greater than or equal to a predetermined threshold δ1, the sample is determined to have a hook effect.

[0026] or,

[0027] The reaction curve was measured between T1 and T2. n Average measurement over the time period Average reference value within the corresponding time period of the reference curve The average response difference Δ% over this time period is calculated using the following formula:

[0028]

[0029] When Δ% is greater than or equal to a predetermined threshold Δ1, the sample is determined to have a hook effect.

[0030] In this specific embodiment, alternatively, in step d) above, the reaction curve is plotted at T... i Measurement value A at time i Reference value A at the corresponding time of the reference curve i(f) The response difference δ is calculated using the following formula. i %:

[0031] δ i % = [(A i -A i(f) ) / A i(f) ]×100%

[0032] When δ i When the percentage is greater than or equal to a predetermined threshold δ2, the sample is determined to have a hook effect, and

[0033] The reaction curve was measured between T1 and T2. n Average measurement over the time period Average reference value within the corresponding time period of the reference curve The average response difference Δ% over this time period is calculated using the following formula:

[0034] Δ% = [(AA (f) ) / A (f) ]×100%

[0035] When Δ% is greater than or equal to a predetermined threshold Δ2, the sample is determined to have a hook effect.

[0036] According to yet another specific implementation, step d) above includes:

[0037] For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) Taking the first derivative with respect to time, we obtain the curves of reaction rate changing with time: v = A' = f'(T) and v' = A (f) '=f'(T (f) );as well as

[0038] In T i The reaction rate value v at time i Compared with the reference reaction rate value v at the corresponding time i(f) The reaction rate difference δv is calculated using the following formula. i %:

[0039] δv i % = (|v i -v i(f) | / v i(f) )×100%

[0040] When δv i When % is greater than or equal to a predetermined threshold δv1, the sample is determined to have a hook effect.

[0041] or,

[0042] In T1~T n Average reaction rate over the time period Compared with the average reference reaction rate value within the corresponding time period The average reaction rate difference Δv% over this time period is calculated using the following formula:

[0043]

[0044] When Δv% is greater than or equal to a predetermined threshold Δv1, the sample is determined to have a hook effect.

[0045] In this specific implementation, step d) may alternatively include:

[0046] For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) Taking the first derivative with respect to time, we obtain the curves of reaction rate changing with time: v = A' = f'(T) and v' = A (f) '=f'(T (f) );as well as

[0047] In T i The reaction rate value v at time i Compared with the reference reaction rate value v at the corresponding time i(f) The reaction rate difference δv is calculated using the following formula. i %:

[0048] δv i % = (|v i -v i(f) | / v i(f) )×100%

[0049] When δv i When % is greater than or equal to a predetermined threshold δv2, the sample is determined to have a hook effect, and

[0050] In T1~T n Average reaction rate over the time period Compared with the average reference reaction rate value within the corresponding time period The average reaction rate difference Δv% over this time period is calculated using the following formula:

[0051]

[0052] When Δv% is greater than or equal to a predetermined threshold Δv2, the sample is determined to have a hook effect.

[0053] In implementations involving first-order differentiation, the actual reaction rate curve may approach or intersect with the reference reaction rate curve. In such cases, the approaching or intersecting regions and their vicinity can be avoided when selecting a timeframe for determining the presence of a hook effect to calculate δv. i %. In one specific implementation, T n Less than the difference δv i The time when % is approximately zero.

[0054] In this specific implementation, step d) above may alternatively include:

[0055] For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) Taking the first derivative with respect to time, we obtain the curves of reaction rate changing with time: v = A' = f'(T) and v' = A (f) '=f'(T (f) );as well as

[0056] When the conditions in T1 to T are simultaneously met nhas an overall tendency of the reaction speed continuously decreasing with time, and the reference reaction speed curve v' = A (f) ' = f'(T (f) ) has an overall tendency of the reference reaction speed first increasing and then decreasing with time, it is determined that the sample has a hook effect.

[0057] According to still another specific embodiment, the step d) comprises:

[0058] deriving the reaction curve A = f(T) and the reference curve A (f) = f(T (f) ) respectively, to obtain curves of reaction acceleration varying with time a = A (f) = A (f) " = f"(T (f) ); and

[0059] the reaction acceleration value a i at T i and the reference reaction acceleration value a i(f) at the corresponding time are calculated to obtain a difference degree of reaction acceleration δa i % according to the following formula:

[0060] δa i % = |(a i - a i(f) ) / a i(f) | x 100%

[0061] When δa i % is greater than or equal to a predetermined threshold value δa1, it is determined that the sample has a hook effect,

[0062] or,

[0063] the average reaction acceleration value a in the time period T1~T n and the average reference average reaction acceleration value a in the corresponding time period are calculated to obtain a difference degree of average reaction acceleration Δa% in the time period according to the following formula:

[0064]

[0065] When Δa% is greater than or equal to a predetermined threshold value Δa1, it is determined that the sample has a hook effect.

[0066] In this specific embodiment, alternatively, the step d) comprises:

[0067] deriving the reaction curve A = f(T) and the reference curve A (f) = f(T(f) By performing second-order differentiation, we obtain the curves of reaction acceleration as a function of time: a = A” = f (T) and a (f) =A (f) =f(T) (f) );and

[0068] In T i The reaction acceleration value a at time i Compared with the reference reaction acceleration value a at the corresponding time i(f) The reaction acceleration difference δa is calculated using the following formula. i %:

[0069] δa i % = |(a i -a i(f) ) / a i(f) |×100%

[0070] When δa i When the percentage is greater than or equal to the predetermined threshold δa2, the sample is determined to have a hook effect, and

[0071] In T1~T n The average reaction acceleration value over the time period Compared with the average reference average reaction acceleration value within the corresponding time period The difference in average reaction acceleration Δa% over this time period is calculated using the following formula:

[0072]

[0073] When Δa% is greater than or equal to the predetermined threshold Δa2, the sample is determined to have a hook effect.

[0074] In this embodiment, T n The value a is less than or equal to the reference reaction acceleration, which reaches a essentially constant value. fconst The moment.

[0075] In the specific implementation of the second-order derivative method described above, the actual reaction acceleration and the reference reaction acceleration may tend to approach each other after a period of reaction, and in some cases, they may even overlap. When selecting the moment for comparing the degree of difference, a time within the time interval before the two acceleration values ​​approach each other can be chosen empirically.

[0076] In this specific implementation, step d) may alternatively include:

[0077] For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) By performing second-order differentiation, we obtain the curves of reaction acceleration as a function of time: a = A” = f (T) and a(f) =A (f) =f(T) (f) );and

[0078] When the conditions in T1 to T are simultaneously met n The reaction acceleration curve a = A” = f (T) over the time period shows an overall trend of increasing reaction acceleration with time, and the reference reaction acceleration curve a (f) =A (f) =f(T) (f) When the overall trend of the reference reaction acceleration decreasing over time is observed, the sample is determined to have a hook effect.

[0079] Furthermore, according to a first aspect of the present invention, a method for identifying the hook effect in immunoturbidimetry is also provided, the method comprising:

[0080] a') After the sample and reaction reagents are mixed to initiate the reaction of the analyte in the sample, the time intervals from T1 to T2 after the reaction begins are obtained. n T over a period of time i Acquire measurement value A at any time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n To generate the reaction curve for that time period;

[0081] b') Differentiate the reaction curve A = f(T) over the time period with respect to time T, and determine whether the sample has a hook effect based on the profile of the differentiated curve.

[0082] According to one implementation, in step a'), T n The time T when the reaction has proceeded to 90% of its total reaction time is less than or equal to the time T. 90 Preferably, the time T is less than or equal to 70% of the total reaction time. 70 More preferably, the time T is less than or equal to 50% of the total reaction time. 50 .

[0083] In this invention, the predetermined time period accounts for 10-70% of the total reaction time, preferably 15-50%, and more preferably 15-40%.

[0084] According to one specific embodiment, the time period for generating the reaction curve of the analyte is from the start of the reaction until the reaction has proceeded to 10-70%, preferably 10-50%, and more preferably 10-40%.

[0085] In this embodiment, the differentiation is either first-order differentiation or second-order differentiation.

[0086] According to a specific embodiment, in step b') the reaction curve A = f(T) is first-order differentiated with respect to time T to obtain a curve of reaction speed as a function of time v = A' = f'(T), and

[0087] When the condition that the reaction speed curve v = A' = f'(T) has an overall tendency of decreasing reaction speed as a function of time is met within the time period T1~T n , it is determined that the sample has a hook effect.

[0088] According to another specific embodiment, in step b') the reaction curve A = f(T) is second-order differentiated with respect to time T to obtain a curve of reaction acceleration as a function of time a = A" = f"(T), and

[0089] When the condition that the reaction acceleration curve a = A" = f"(T) has an overall tendency of increasing reaction acceleration as a function of time is met within the time period T1~T n , it is determined that the sample has a hook effect.

[0090] According to the method for identifying hook effect of the present application, when the profile of the curve after differentiation meets the predetermined characteristics, it is identified that the sample has a hook effect, and the detection is terminated.

[0091] In the method of the present application, the sample is whole blood sample of a mammal, preferably a human.

[0092] In the method of the present application, the analyte in the sample is a protein, preferably C-reactive protein or serum amyloid protein.

[0093] The method of the present application further comprises, before step a) or a'), a step of drawing the sample into a reaction chamber and adding a reaction reagent mixture, the sample reacts with the reagent, and the red blood cells in the sample are lysed.

[0094] In the method, the reaction reagent comprises a substance capable of specifically reacting with the analyte, preferably a latex coated with the specific reaction substance.

[0095] According to an embodiment, before step a) or a'), the sample is aliquoted into at least two aliquots, one of which is subjected to routine blood test, and the other of which is subjected to the analyte test and subjected to the steps of the aforementioned method.

[0096] According to a second aspect of the present application, a blood analysis system is provided, the blood analysis system comprising:

[0097] a sampling unit for obtaining a blood sample and delivering the blood sample to the reaction unit;

[0098] A reagent supply unit is used to store the first reaction reagent and supply it to the reaction unit as needed;

[0099] The reaction section includes a first reaction chamber for mixing the blood sample with the first reaction reagent to prepare a first test solution;

[0100] The detection system includes a light source and a detector for detecting the first test solution, used to acquire the measured value of the test solution;

[0101] A controller is coupled to the sampling unit, reagent supply unit, reaction unit, and detection system, and controls the operation of the sampling unit, reagent supply unit, reaction unit, and detection system;

[0102] The processor, which is coupled to the detection system,

[0103] The processor receives the reaction start time T1~T1 from the detection system. n T within the scheduled time period i Measurement value A at time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n To generate a reaction curve for that time period; to estimate the concentration Ce of the analyte based on the reaction curve for that time period and a pre-stored calibration curve; and to determine the concentration Ce of the analyte based on the estimated concentration Ce. e Obtain a pre-stored reference curve for that concentration; and compare the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period to determine whether the sample exhibits a hook effect.

[0104] Alternatively, the processor obtains the reaction start time T1 to T2 from the detection system. n T over a period of time i Acquire measurement value A at any time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n The processor generates a response curve for that time period; it differentiates the response curve A = f(T) with respect to time T, and determines whether the sample has a hook effect based on the curve profile after differentiation. The processor then outputs the determination result to the controller.

[0105] According to one embodiment, the step of the processor comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period includes: comparing the measured value of the reaction curve for the predetermined time period with the measured value of the reference curve for the corresponding time period.

[0106] According to another embodiment, the step of comparing the distribution information of the reaction curve of the predetermined time period with the distribution information of the reference curve of the corresponding time period by the processor comprises: first-order derivation of the reaction curve and the reference curve to obtain a reaction velocity curve and a reference velocity curve, and comparison of the reaction velocity curve and the reference velocity curve.

[0107] According to another embodiment, the step of comparing the distribution information of the reaction curve of the predetermined time period with the distribution information of the reference curve of the corresponding time period by the processor comprises: second-order derivation of the reaction curve and the reference curve to obtain a reaction acceleration curve and a reference acceleration curve, and comparison of the reaction acceleration curve and the reference acceleration curve.

[0108] According to another embodiment, the step of comparing the distribution information of the reaction curve of the predetermined time period with the distribution information of the reference curve of the corresponding time period by the processor comprises: second-order derivation of the reaction curve and the reference curve to obtain a reaction acceleration curve and a reference acceleration curve, and comparison of the reaction acceleration curve and the reference acceleration curve.

[0109] The blood analysis system of the present application, when the controller receives the result that the current detection sample has hook effect, controls the detection system to stop detection;

[0110] The controller controls the sampling unit to sample the sample again and deliver the sample to the first reaction chamber of the reaction unit;

[0111] The controller controls the reagent supply unit to provide the first reaction reagent to the first reaction chamber to prepare a second test solution, and the dilution multiple of the sample in the second test solution is greater than that in the first test solution; and

[0112] The controller controls the detection system to detect the second test solution.

[0113] The detector can include a photometer, specifically a turbidimeter and / or a nephelometer.

[0114] The blood analysis system of the present application, wherein the sample is a whole blood sample, and the first reagent includes a hemolytic agent for lysing red blood cells in the sample and a latex reagent for immunoturbidimetric reaction with the analyte in the sample.

[0115] The blood analysis system further comprises a second detection system, and the reaction unit further comprises a second reaction chamber; the second detection system comprises a light source, a flow chamber for cell-by-cell queuing, a liquid path system, and a second detector.

[0116] The controller controls the sampling unit to divide the sample into two parts, which are respectively delivered to the first reaction chamber and the second reaction chamber; controls the reagent supply unit to deliver the second reagent to the second reaction chamber, and the sample reacts with the second reagent in the second reaction chamber to obtain a third test solution; controls the second detection system, under the driving of the liquid path system, the third test solution is delivered to the flow chamber, the light source irradiates the flow chamber, and the second detector collects the scattering light signal generated by the cells.

[0117] The processor obtains the scattering light signal, and divides the white blood cells in the sample into at least three types of lymphocytes, monocytes and neutrophils according to the scattering light signal.

[0118] Further, the second detection system further comprises a third detector, which collects a fluorescence signal generated by the cells, and divides the white blood cells into at least four types of lymphocytes, monocytes, neutrophils and eosinophils according to the scattering light signal and the fluorescence signal.

[0119] According to a specific embodiment, the second reagent comprises a hemolytic agent and a staining agent. According to a third aspect of the present application, a computer readable medium is provided, which stores executable instructions, wherein the computer readable storage medium is configured to cause a processor to execute the executable instructions to implement the hook effect identification method for immunoturbidimetry.

[0120] The present application utilizes the high dependence of the reaction degree of the antigen-antibody initial reaction stage on the reactants, acquires the reaction information of a certain stage, especially the initial stage, and utilizes the reaction information of the initial stage to judge whether the sample has hook effect, which greatly improves the alarm speed of the abnormal sample with hook effect, thereby shortening the detection time of the sample with hook effect. It has positive significance for improving the speed of outpatient examination, especially the speed of protein detection in blood at the same time as blood routine. BRIEF DESCRIPTION OF DRAWINGS

[0121] Figure 1 A schematic diagram for plotting the amount of antibody-antigen precipitation relative to the amount of antigen to show hook effect;

[0122] Figure 2 A flowchart according to the first embodiment of the present application;

[0123] Figure 3 A comparison diagram of the reaction curve with hook effect and the reference curve according to an embodiment of the present application;

[0124] Figure 4 A comparison diagram of the reaction curve with hook effect and the reference curve according to an embodiment of the present application; Figure 3A comparison of the reaction rate versus time curves obtained by taking the first derivative of the reaction curve and the reference curve with respect to time.

[0125] Figure 5 According to one embodiment of the present invention, for Figure 3 A comparison of the reaction acceleration versus time curves obtained by taking the second derivative of the reaction curve and the reference curve with respect to time.

[0126] Figure 6 A flowchart of the second embodiment of the present invention;

[0127] Figure 7 This is a schematic diagram of a blood analysis system according to one embodiment;

[0128] Figure 8 The actual reaction curves and corresponding reference reaction curves for the five samples tested according to Example 1 are shown in the figure.

[0129] Figure 9 The graph shows the changes in actual reaction rate and reference reaction rate after taking the first derivative of the actual reaction curves and the corresponding reference reaction curves for the five samples tested according to Example 1; and

[0130] Figure 10 The graph shows the changes in actual reaction acceleration and reference reaction acceleration after taking the derivatives of the actual reaction curves and the corresponding reference reaction curves of the five samples tested according to Example 1. Detailed Implementation

[0131] The technical solutions of the present invention will be clearly and completely described below with reference to specific embodiments and accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0132] Throughout this specification, unless otherwise specified, the terminology used herein should be understood as having the meaning commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of any conflict, the definitions set forth herein shall prevail.

[0133] The term "immunoturbidimetric assay" is a method for determining the concentration of an analyte by measuring the change in intensity of transmitted light (i.e., transmission method) or by measuring the change in intensity of scattered light at a predetermined angle to the incident light (i.e., scattering method) based on particles suspended in the reaction system formed by an immunoagglutination reaction.

[0134] The method of the present application for recognizing the hook effect is applicable to any immunonephelometry. A particularly preferred method is latex-enhanced nephelometry.

[0135] In latex-enhanced nephelometry, the antibody corresponding to the analyte is coated on latex particles of about 15-60 nm in size to increase the volume of the antigen-antibody conjugate, thereby enhancing the change in the intensity of transmitted or scattered light to increase the detection sensitivity. Commonly used latex particles are polymer particles such as polystyrene.

[0136] The specific assay types of immunonephelometry are generally end-point assay and rate assay, or a modified method combining both. The method of the present application for recognizing the hook effect is applicable to any type of assay method without particular limitation.

[0137] As will be described in detail below, the method of the present application is suitable for quickly determining whether a sample has the hook effect in any immunonephelometry and promptly terminating the detection of a sample having the hook effect. For a sample without the hook effect, the method of the present application does not affect or change the normal detection procedure.

[0138] The hook effect recognition method of the present application generates a reaction curve using the light signal (e.g., light intensity or absorbance) measured in a predetermined time period of the immunoreaction of the analyte in the sample, and determines whether the sample has the hook effect (HOOK effect) using the distribution information of the reaction curve in the predetermined time period. In the method of the present application, only the measurement information in a small predetermined time period of the entire reaction time is used to determine whether the hook effect exists.

[0139] According to a first embodiment of the present application, after obtaining the reaction curve in a predetermined time period, the concentration of the analyte is estimated, the reaction curve corresponding to the estimated analyte concentration in the normal reaction state is obtained as a reference curve, and whether the sample has the hook effect is determined by comparing the distribution information of the reaction curve of the analyte in the predetermined time period with that of the reference curve.

[0140] The "reference curve" in the present context means the reaction curve of the analyte in a normal sample without the hook effect at the estimated concentration of the analyte from the measurement values in the predetermined time period.

[0141] According to one embodiment, the period of time is a predetermined time period from T1 to T2 after the start of the reaction. According to a specific embodiment, T1 = 0, i.e., the period of time is from the start of the reaction to a certain time T2. According to another specific embodiment, T1 > 0, i.e., the period of time is from a certain time T1 after the start of the reaction to another time T2. n n n

[0142] ​​​According to the present application, the predetermined time period is a time period in the first 90% of the reaction. That is, the time corresponding to the point at which the reaction has proceeded to 90% of the total reaction time is T 90 , then T n ≤ T 90 . For example, the predetermined time period can be a time period in the first 80%, first 70%, first 50%, first 30%, or even first 10% of the total reaction.

[0143] When the reaction signal data of the immune reaction or the set obtained by derivation of the reaction signal data has a maximum value in the middle of the curve of the reaction time (i.e., when the trend of the slope of the reaction curve changes), for example, when the maximum value appears in the middle of the reaction rate reference curve, the predetermined time period is a time period before the maximum value of the total reaction. The portion of the total reaction process selected as the predetermined time period varies depending on the analyte. It is generally determined empirically according to the specific circumstances of the determination. According to the present application, the predetermined time period is preferably a time period at the beginning of the reaction, i.e., T1= 0. According to this preferred mode, it is advantageous to identify the sample with a hook effect as soon as possible and to stop the detection.

[0144] In general, the predetermined time period for generating the reaction curve of the analyte is 10% to 70% of the total reaction time, such as 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or 60% of the total reaction time. According to a preferred embodiment, the time period for generating the reaction curve of the analyte is 15% to 50%, more preferably 15% to 40% of the total reaction time.

[0145] According to the most preferred embodiment, the time period for generating the reaction curve of the analyte is the reaction curve in the first period after the beginning of the reaction (i.e., T1= 0), and the end point Tn can be a time period of 10-70% of the reaction (i.e., T n = T 10-70 , preferably 10-50% (i.e., T n = T 10-50 , and more preferably 10-40% (i.e., T n = T 10-40 .

[0146] According to the method of the present application, within the above-mentioned time period, the actual reaction curve is obtained based on the actually detected information, such as absorbance, and the estimated analyte concentration is obtained based on the stored standard curve. The method of estimating the analyte concentration is the same as the conventional method of obtaining the analyte concentration, with the difference being that in the method of the present application, only the detection information of a portion of the reaction, particularly the beginning of the reaction, is used to estimate the concentration of the analyte.

[0147] Further, the method of the present application utilizes the estimated analyte concentration to obtain a corresponding reference curve. As mentioned before, by comparing the distribution information of the actual reaction curve with the reference reaction curve, it is possible to quickly determine whether the sample has a hook effect or not.

[0148] According to the method of the present application, the identification of the hook effect is performed by means of the reaction curve obtained within a relatively short predetermined time period, so that it is possible to identify whether the hook effect exists or not within a short time after the start of the reaction, and further, it is possible to terminate the detection as early as possible in the case where the hook effect exists.

[0149] In the case where it is identified that the sample has a hook effect, the detection is terminated and the reaction chamber is cleaned or the sample is re-prepared or taken in by using another reaction chamber and re-detected after being appropriately diluted.

[0150] In the case where the sample does not have a hook effect, the reaction is continued to be completed and the detection result is reported.

[0151] The method of the present application is particularly advantageous in obtaining the detection result as early as possible in the case of whole blood detection. In the case of clinical blood detection in a hospital, blood routine detection is the most common detection item. A conventional fully automatic blood detection apparatus can quickly complete each detection item such as the determination of red blood cells, white blood cells and platelets, and white blood cell classification in blood routine detection. However, the detection of specific proteins in blood is relatively slow. In most cases, the detection time of CRP, for example, determines the speed of the examination in the clinic.

[0152] In the conventional method, if the sample has a hook effect (which is often the case in patients), it is not possible to determine this until the first detection is completed, and the sample is re-processed and re-detected. This further affects the speed of the examination in the clinic. Therefore, the method of the present application is advantageous in speeding up the examination in the clinic. For example, if the determination time of CRP is 60 s, the method of the present application can complete the identification within 10 to 20 s.

[0153] The detection apparatus for the immunoturbidimetric method can be a turbidimeter or a nephelometer, specifically, a scatterometer or a transmittance detector. The measurement can be performed in the range of ultraviolet or near ultraviolet to visible light (typically, 300 to 800 nm). The temperature of the reaction system during the measurement is typically ambient temperature, for example, 20 to 40°C, and preferably about 37°C.

[0154] The term "sample" or "analyte sample" generally refers to a blood sample, particularly a whole blood sample, in the context. In this context, the analyte sample is typically a peripheral blood or venous blood sample derived from a mammal, and particularly a human blood sample. The sample has been subjected to necessary treatment before the immunoreaction. The treatment includes, but is not limited to, anticoagulation treatment, dilution treatment, hemolysis treatment and the like.

[0155] When the immunoturbidimetry is performed, the sample is first sucked from the sample tube into the reaction chamber, and the reaction reagents are added in sequence or simultaneously, the reaction is started, and the detection is started simultaneously. The reaction reagents are reagents that can react with the analyte in the art, such as latex coated with antibodies or antigens, and reagents for aggregation of immune complexes, such as polyethylene glycol. Details are not repeated here. The reaction reagent can be a solution, or a plurality of solutions, such as a hemolytic agent for dissolving red blood cells in a whole blood sample, a polyethylene microsphere latex reagent coated with antibodies, a buffer for diluting the whole blood sample, etc.

[0156] The term "analyte" herein refers to an antigen, an antibody, a protein, a polypeptide, etc. in a sample, especially a blood sample. Specifically, it can be, but is not limited to, C-reactive protein (CRP), serum amyloid A (SAA), procalcitonin (PCT), interleukin-6 (IL-6), human chorionic gonadotropin, growth hormone, luteinizing hormone, alpha-fetoprotein, and carcinoembryonic antigen, etc.

[0157] The following references are incorporated by reference in their entirety: Figure 2 The first embodiment of the present application is further described in detail. The method of the present application comprises the following steps. In step S101, the sample is first mixed with the reaction reagent to start the reaction of the analyte in the sample; then, the measurement value A n at time T i in a predetermined time period T i is obtained, where i is an integer from 1 to n, thereby obtaining a plurality of measurement values A1-A n to generate a reaction curve of the time period. Next, in step S102, the concentration C e of the analyte is estimated based on the reaction curve of the time period and the pre-stored calibration curve. In step S103, the reference curve of the concentration C e is obtained according to the estimated concentration C d of the analyte. Finally, in step S104, the distribution information of the reaction curve of the time period is compared with the distribution information of the reference curve of the corresponding time period. In step S105, it is determined whether the predetermined characteristics are met (e.g., the difference between the distribution information of the reaction curve and the reference curve is higher than a predetermined threshold, or the change trend of the distribution information of the two curves meets a certain rule), and if so, the hook effect is identified, and the detection is terminated in step S106; otherwise, the detection is continued in step S107.

[0158] Specifically, the reaction rate R d is calculated based on the reaction curve of the predetermined time period, the concentration C d of the analyte is estimated according to the reaction rate R e , and the reaction rate R d is the reaction rate at the end of the time period (Tn ) the measured value (A n ) and the measured value (A1) at the start of the time period (T1) (A n -A1) : R d = A n -A1.

[0159] Applicants have found that for samples without hook effect, the reaction rate in the initial period of the reaction is consistent with the calibration curve, and thus the estimated concentration of the analyte is close to the concentration measured at the end of the reaction. The reference curve obtained from the estimated concentration in the normal reaction state (without hook effect) is consistent with or close to the reaction curve of the initial reaction of the sample.

[0160] On the contrary, for samples with hook effect, the concentration of the analyte in the sample exceeds the linear part of the calibration curve. Thus the estimated concentration of the analyte from the reaction curve at the initial reaction is significantly higher, and thus the reference curve further obtained is deviated from the reaction curve of the sample with hook effect (see Figure 3 , where a comparison chart of the reaction curve of the sample with hook effect and the reference curve in the initial period of the reaction is shown).

[0161] Similarly, the reaction rate and acceleration of the sample with hook effect are significantly different from those of the normal reaction represented by the reference curve (see Figure 4 and Figure 5 ).

[0162] According to one specific embodiment of the present embodiment, when the difference between the distribution information of the reaction curve and the reference curve in the time period of T1~T n is below a predetermined threshold, it is identified as no hook effect; and when the difference between the distribution information of the reaction curve and the reference curve in the time period of T1~T n is above a predetermined threshold, it is identified as hook effect.

[0163] In the present application, the reaction time and reaction distribution information under the determination conditions are applied to the method for identifying hook effect. The reaction distribution information includes the characteristics of the measured value (such as absorbance), the characteristics of the reaction rate and / or the characteristics of the reaction acceleration.

[0164] The characteristics of the measured value can be directly the optical measurement value (such as an absorbance value) in a predetermined time period, or the average measured value in a predetermined time period, or both.

[0165] According to one specific embodiment, in the judging step, the optical measurement value A i of the reaction curve at a time T ithe reference value A at the corresponding time of the reference curve i(f) the reaction difference δ% calculated according to the following formula i

[0166] δ i % = [(A i -A i(f) ) / A i(f) ] x 100%.

[0167] When the reaction difference δ i % is greater than or equal to a predetermined threshold value δ1, it is determined that the sample has a hook effect.

[0168] According to another specific embodiment, the average measurement value of the reaction curve A = f(T) in the time period T1~T n the average reference value A of the reference curve in the corresponding time period the average reaction difference Δ% in the time period is calculated according to the following formula:

[0169]

[0170] When the average reaction difference Δ% is greater than or equal to a predetermined threshold value Δ1, it is determined that the sample has a hook effect.

[0171] According to still another embodiment, the reaction difference δ i % and the average reaction difference Δ% are calculated according to the above embodiments. When the reaction difference δ i % is greater than or equal to a predetermined threshold value δ2, and the average reaction difference Δ% is greater than or equal to a predetermined threshold value Δ2, it is determined that the sample has a hook effect.

[0172] The distribution information can also be a value of a reaction speed or a reaction acceleration in a predetermined time period, or a value of an average reaction speed or an average reaction acceleration in a predetermined time period.

[0173] Specifically, the reaction curve A = f(T) and the reference curve A f = f(T f ) are respectively first-order differentiated with respect to time, to obtain the curves of the reaction speed v = A' = f'(T) and v' = A (f) ' = f'(T (f) ) changing with time.

[0174] the reaction speed value v i at T i and the reference reaction speed value v fi(f) are calculated according to the following formula to obtain the reaction speed difference δv i %:

[0175] δv​​i % = (|v i -v i(f) | / v i(f) )×100%.

[0176] When δv i When the percentage is greater than or equal to the predetermined threshold δv1, the sample is determined to have a hook effect.

[0177] Or, in T1 to T n Average reaction rate over the time period Compared with the average reference reaction rate value within the corresponding time period The average reaction rate difference Δv% over this time period is calculated using the following formula:

[0178]

[0179] When Δv% is greater than or equal to a predetermined threshold Δv1, the sample is determined to have a hook effect.

[0180] According to another embodiment, the reaction difference δv is calculated according to the above embodiment. i % and average reaction difference Δv%. When the reaction difference δv i When the percentage of the mean response difference Δv% is greater than or equal to the predetermined threshold δv2, the sample is determined to have a hook effect.

[0181] In the reaction rate reference curve, the reaction rate between the analyte and the detection reagent initially increases and then decreases, with a maximum value point on the curve. That is, during the detection process, a maximum reaction rate occurs in the middle of the reaction (see...). Figure 4 ).

[0182] However, in samples exhibiting the hook effect, the reaction rate between the analyte and the detection reagent generally decreases continuously. The actual reaction rate is much greater than the reference reaction rate in the early stage of the reaction, but often less than the reference reaction rate in the later stage.

[0183] Based on a specific example, the time T selected for comparing the degree of difference... i Or a certain time period T1 to T n Preferably, at the time point corresponding to the occurrence of the maximum reference reaction rate on the reference reaction rate curve (i.e., T... max Selected beforehand.

[0184] Furthermore, the study found that in samples exhibiting the hook effect, the reaction rate curve intersects with the reference reaction rate curve. In this case, a specific time T is selected for comparing the degree of difference. i, the time point corresponding to the intersection point (i.e. actual reaction rate = reference reaction rate) should be avoided, and preferably, a time point before the corresponding time point is selected. When a time period T1~T n may also be selected.

[0185] The shapes of the reaction curves and their reference curves of different samples may be different, and those skilled in the art can understand that samples known to have no hook effect and samples known to have hook effect can be counted to obtain appropriate values of T i or T1, T n according to the above principles.

[0186] The reaction acceleration can be obtained by second-order derivation. Specifically, the second-order derivation is performed on the reaction curve A = f(T) and the reference curve A (f) = f(T (f) ) respectively, to obtain the curves of the reaction acceleration changing with time a = A” = f”(T) and a (f) = A (f) ” = f”(T (f) ).

[0187] The reaction acceleration value a i at T i and the corresponding reference reaction acceleration value a i(f) are calculated to obtain the reaction acceleration difference degree δa i % according to the following formula:

[0188] δa i % = |(a i -a i(f) ) / a i(f) | × 100%

[0189] When δa i % is higher than a predetermined threshold δa1, it is determined that the sample has hook effect.

[0190] Alternatively, the average reaction acceleration value in the time period T1~T n and the average reference average reaction acceleration value a (f) in the corresponding time period are calculated to obtain the average reaction acceleration difference degree Δa% in the time period according to the following formula:

[0191]

[0192] When Δa% is higher than a predetermined threshold Δa1, it is determined that the sample has hook effect.

[0193] According to still another embodiment, the reaction difference degree δa i% and average reaction difference Δa%. When the reaction difference δa i When the percentage of response difference Δa% is greater than or equal to a predetermined threshold δa2, and the average response difference Δa% is greater than or equal to the predetermined threshold Δa2, the sample is determined to have a hook effect. In this example, the predetermined time period is preferably selected as the plateau period a of the response acceleration of the reference curve. const(f) (If it exists) the previous time period (see Figure 5 ).

[0194] The aforementioned predetermined thresholds can be the same or different. They vary depending on the detection equipment, detection method, sample, and analyte used. They are usually statistical or empirical values, and are T. n The function.

[0195] According to another embodiment, the distribution information may also be a contour line showing the change of reaction speed or reaction acceleration over time within a predetermined time period, and the presence of a hook effect in the sample may be determined based on the trend of the change of the contour line.

[0196] Specifically, see Figure 4 In T1 to T n Within a given time period, the reaction rate curve shows an overall trend of decreasing reaction rate over time, while the reference reaction rate curve shows an overall trend of first increasing and then decreasing the reference reaction rate over time. If the above characteristic profile exists, it can be determined that the sample exhibits a hook effect.

[0197] Or, in other words, when simultaneously satisfying conditions from T1 to T... n The smoothed reaction rate curve at two different times T within the time period i and T j The reaction rate value satisfies v i >v j (where j is an integer from 2 to n, and j > i), and there exists a maximum value v on the fitted reference reaction rate curve within the corresponding time period. max(f) When the sample is found to have a hook effect, it is determined that the sample has a hook effect.

[0198] Or see Figure 5 When simultaneously satisfying conditions T1 to T n Within the time period, most of the two adjacent times T i and T j The reaction acceleration value satisfies a i <a j And in the corresponding T f1 ~T fn Within the time period, most adjacent time intervals T fi and T fj The reference reaction acceleration value satisfies a fi >afj When j is an integer from 2 to n and j > i, it is determined that the sample has a hook effect.

[0199] According to the second embodiment of the present application, after obtaining the reaction curve of the analyte in the sample under test within a predetermined time period from the start of the reaction, the profile of the reaction rate or the reaction acceleration varying with time within the predetermined time period is further obtained from the reaction curve, and it is determined whether the sample has a hook effect according to the variation trend of the profile. In this embodiment, the analyte concentration does not need to be estimated, and the reference curve is obtained.

[0200] In this embodiment, the sample, the analyte and the predetermined time period are defined as described above. The reaction curve within the predetermined time period is obtained in the same way.

[0201] Reference Figure 6 The second embodiment is described in detail. According to this embodiment, in step S201, after the sample is mixed with the reaction reagent to start the reaction of the analyte in the sample, the measurement value A is obtained at time T n from T i to T i , where i is an integer from 1 to n, so as to obtain a plurality of measurement values A1 to An, to generate the reaction curve of the time period. In step S202, the reaction curve A = f(T) within the time period is differentiated with respect to time T, where the first-order differentiation obtains the reaction rate curve, and the second-order differentiation obtains the reaction acceleration curve. Then, the judgment step S203 is performed to determine whether the sample has a hook effect according to the profile of the curve after differentiation. When there is a hook effect, step S204 is performed to terminate the detection; otherwise, step S205 is performed to continue the current detection.

[0202] Specifically, the reaction curve A = f(T) is first-order differentiated with respect to time T to obtain the curve v = A' = f'(T) of the reaction rate varying with time, and when it is satisfied that, within the time period from T n to T i , most of the reaction rate values of adjacent two time points T j and T i satisfy v j > v n , where j is an integer from 2 to n and j > i, it is determined that the sample has a hook effect.

[0203] According to another specific embodiment, the reaction curve A = f(T) is second-order differentiated with respect to time T to obtain the curve a = A" = f"(T) of the reaction acceleration varying with time, and when it is satisfied that, within the time period from T i to T jthe reaction acceleration value of the sample satisfies a i <a j wherein j is an integer from 2 to n, and i < j, it is determined that the sample has hook effect.

[0204] When it is identified that the sample has hook effect, the detection is terminated; otherwise, the detection is continued.

[0205] Optionally, the sample is re-prepared or re-pipetted into the reaction chamber, and is re-measured after being properly diluted at an increased dilution ratio.

[0206] The present application also relates to a blood analysis system capable of implementing the above-identified method.

[0207] The blood analysis system comprises a sampling part, a reagent supply part, a reaction part comprising a reaction chamber, a data processing module and a controller.

[0208] Referring to Figure 7 wherein a specific blood analysis system, a fully automatic blood analyzer 1 is schematically shown. The blood analyzer 1 has a first casing 11, a second casing 12, a sampling part 20, a reaction part 30, a reagent supply part (not shown), a detection system 40, a main system 50 and an output part 60. In actual application, the output part 60 can be a user interface. In the present embodiment, the detection system 40 and the main system 50 are arranged inside the second casing 12. The reaction part 30 is arranged inside the first casing 11. The detection system is close to the reaction chamber (not shown) of the reaction part 30 for facilitating detection. The output part 60 and the sampling part 20 are on the outer surface of the first casing 11.

[0209] The sampling part 20 has a sampling needle, which collects a blood sample and delivers the collected blood sample to the reaction chamber of the reaction part 30.

[0210] The reagent supply part stores the first reaction reagent for reacting with the blood sample as described above and other necessary reagents, and supplies the corresponding reagents to the reaction part as needed.

[0211] The reaction part 30 can comprise a first reaction chamber, which is configured to allow the blood sample from the sampling part and the first reaction reagent from the reagent supply part to react in the reaction chamber to generate antibody-antigen complex particles.

[0212] The detection system 40 comprises a light source and a detector for detecting the reaction system in the reaction chamber to obtain a measurement value of the reaction system. The detector can comprise a turbidimeter and / or a nephelometer.

[0213] The host system 50 comprises a processor, a memory and a controller. The controller is coupled with the sampling unit, the reagent supply unit, the reaction unit and the detection system, and controls the actions of the sampling unit, the reagent supply unit, the reaction unit and the detection system. The processor is coupled with the detection system 50. The memory is a non-transitory computer readable storage medium, and stores a computer program therein. When the computer program is executed by the processor, the above-mentioned method for identifying the hook effect is implemented. The controller is also operatively connected with the processor, and is configured to receive the detection result from the processor.

[0214] In the present application, when the processor determines that the sample has the hook effect, the controller controls the detection system to stop the detection.

[0215] Further, the system of the present application can also automatically perform re-detection. The controller further sends a re-detection instruction. Under the control of the controller, the liquid in the reaction chamber is removed and the reaction chamber is cleaned; the sampling unit 20, the reaction unit 30, the reagent supply unit and the detection system 40 are again matched to perform re-detection. The sampling unit is controlled to sample the sample again, and the blood sample is transported to the first reaction chamber of the reaction unit; the reagent supply unit is controlled to provide the first reaction reagent to the first reaction chamber to prepare a second test solution, the dilution multiple of the sample in the second test solution being greater than that in the first test solution; and the detection system is controlled to perform re-detection on the second test solution.

[0216] The output unit 60 is configured to output the detection result of the analyte in the sample according to the instruction sent by the controller when the processor determines that the sample does not have the hook effect.

[0217] The blood analysis system of the present application can also have an alarm device (not shown) for alarming when the data processing module determines that the sample has the hook effect.

[0218] According to a further embodiment, the blood analysis system of the present application further comprises a second detection system, and the reaction unit further comprises a second reaction chamber.

[0219] The second detection system comprises a light source, a flow chamber for the cells to pass through one by one, a liquid path system and a second detector.

[0220] According to this embodiment, the blood analysis system of the present application can simultaneously perform routine blood test on the whole blood sample and perform immunoturbidimetry test on specific proteins (e.g., C-reactive protein).

[0221] In this embodiment, the controller controls the sampling unit to divide the sample into two portions, which are transported to the first reaction chamber and the second reaction chamber, respectively.

[0222] The detection by the immunoturbidimetry is as described above. In another detection, such as the detection in blood routine test, the controller controls the reagent supply part to deliver the second reagent to the second reaction chamber. The sample reacts with the second reagent in the second reaction chamber to obtain a third test solution.

[0223] An example of the second reagent can include a hemolytic agent and a staining agent, but is not limited thereto.

[0224] The second detection system is controlled, under the driving of the liquid path system, to deliver the third test solution to the flow chamber, the light source irradiates the flow chamber, and the second detector collects the scattering light signal generated by the cells.

[0225] The processor obtains the scattering light signal and classifies the white blood cells in the sample into at least three types of lymphocytes, monocytes and neutrophils according to the scattering light signal.

[0226] The second detection system further includes a third detector that collects a fluorescent signal generated by the cells, and classifies the white blood cells into at least four types of lymphocytes, monocytes, neutrophils and eosinophils according to the scattering light signal and the fluorescent signal.

[0227] According to the scattering light and the fluorescent signal collected by the second detection system, the counts and classifications of red blood cells, white blood cells, platelets and the like can be further performed.

[0228] The application further provides a computer readable storage medium. The computer readable storage medium stores executable instructions, which, when executed by the processor, implement the steps of the aforementioned method for recognizing the hook effect of the immunoturbidimetry. The computer readable storage medium can be the aforementioned memory or a component thereof, which stores the computer program and is executed by the processor of the blood analysis system to complete the aforementioned method steps.

[0229] The computer readable storage medium can be an FRAM, a ROM, a PROM, an EPROM, an EEPROM, a Flash Memory, a magnetic surface memory, an optical disc or a CD-ROM, etc., or can be various devices including one or any combination of the above storage media.

[0230] The application will be further described below through specific examples.

[0231] Example 1

[0232] Five whole blood samples known to have hook effect were detected on Mindray BC-5390 CRP in the detection of serum amyloid A (SAA). The longest detection time of each sample was 80 s. The signal per second was obtained to draw the original reaction curve (i.e. absorbance curve, see Figure 8 ).

[0233] Obtain the signal values A1 and A2 at T1=5s and T2=20s on the original reaction curve. Calculate the signal mean value using the signal value (i.e. absorbance) per second between T1=5s and T2=20s Obtain the signal values A1 and A2 at T1=5s and T2=20s on the original reaction curve. Calculate the signal mean value using the signal value (i.e. absorbance) per second between T1=5s and T2=20s e (see Table 1 below). Then, according to the estimated concentration C e , find the corresponding curve from the stored reference curves, and take this curve as the reference reaction curve (see e Figure 8 ).

[0234] Similarly, obtain the signal values A 1(f) and A 2(f) at T1 and T2 on the reference reaction curve. Calculate the signal mean value using the signal value per second between T1 and T2 (see Table 2 below).

[0235] Table 1: Original measurement results of SAA in samples and estimated concentrations

[0236]

[0237] Table 2: Corresponding signal values and signal mean values at T1=5s and T2=20s according to the reference reaction curve

[0238]

[0239] Calculate the difference degree of the signal values at T1 and T2 time points on the original reaction curve and the reference reaction curve according to the following formula:

[0240] δ i % = [(A i - A i(f) ) / A i(f) ] x 100%

[0241] T i = 5s or 20s, when δ i % ≥ 15%, it is determined that there is a hook effect;

[0242] In this embodiment, δ i % at 5s and 20s are both greater than 15%. In some cases where the alarm accuracy requirement is not high, it can be determined according to whether δ i % at 5s or 20s is greater than a threshold value to alarm whether the sample has a hook effect. Alternatively, it can be determined according to whether δ i % at other time points between T1 and T2, such as 10s or 15s, is greater than a threshold value to alarm. ​

[0243] Alternatively, the difference between the original reaction curve and the reference reaction curve 5s~20s signal average is calculated according to the following formula:

[0244]

[0245] When Δ%≥15%, it is determined to have a hook effect.

[0246] The specific results are shown in Table 3.

[0247] Table 3: Signal value difference and 5~20s signal average difference when T1=5s, T2=20s 16

[0248]

[0249] From the above Table 3, it can be seen that the corresponding reference curve is found from the stored calibration curve according to the estimated SAA concentration within the first 15s of the reaction, and the difference between the original signal value at a certain time within the first 15s of the reaction and the signal value at the corresponding time of the reference curve is compared. When the difference is greater than or equal to 15%, it is determined that the sample has a hook effect.

[0250] Alternatively, the average of the original signal within the first 15s of the reaction and the average of the reference curve at the corresponding time period are compared. When the difference is greater than or equal to 15%, it is determined that the sample has a hook effect.

[0251] Example 2

[0252] The original reaction curve and the reference reaction curve obtained in Example 1 are respectively subjected to first-order derivation to obtain the original reaction velocity curve and the reference reaction velocity curve (see Figure 9 ).

[0253] According to the original reaction velocity curve, the original reaction velocities v1 and v2 at T1=5s and T2=20s are obtained. The average of the reaction velocity per second between T1 and T2 is calculated according to (see Table 4 below).

[0254] Similarly, the reference reaction velocities v 1(f) and v 2(f) at T1 and T2 after the first-order derivation of the reference reaction curve are obtained. The average of the reaction velocity per second between T1 and T2 is calculated according to (see Table 5 below).

[0255] Table 4: Original reaction velocity and average reaction velocity of SAA in the sample at T1 and T2

[0256]

[0257] ​Table 5: Reaction rate at Tl and T2 and average reaction rate in the reference reaction rate curve

[0258]

[0259] The difference of reaction rate at Tl and T2 in the original reaction rate curve and the reference reaction rate curve is calculated according to the following formula:

[0260] δv i %= (|v i -v i(f) | / v i(f) ) x 100%

[0261] T i = 5s or 20s, when δv i %≥ 12% at one of Tl and T2, it is determined that the sample has hook effect.

[0262] The skilled in the art can understand that other time points between Tl and T2, such as 10s or 15s, can also be selected to determine whether δv i % is greater than the threshold value to alarm. In addition, in the present embodiment, since the original reaction rate curve and the reference reaction rate curve have time points close to each other or even cross, the skilled in the art can understand that it is best to avoid selecting these time points to calculate δv i %. Different time points can also have different thresholds, and the appropriate threshold value can be obtained according to the statistics of known samples.

[0263] Or the difference of average reaction rate between 5s and 20s in the original reaction rate curve and the reference reaction rate curve is calculated according to the following formula:

[0264]

[0265] When Δv%≥ 12%, it is determined that the sample has hook effect.

[0266] The specific results are shown in Table 6.

[0267] Table 6: Difference of reaction rate at Tl = 5s and T2 = 20s and difference of average reaction rate between 5s and 20s

[0268]

[0269] As can be seen from Table 6 above, the reaction rate of the sample within the first 15s has a significant difference with the reaction rate of the reference reaction curve. The difference of reaction rate at a certain time point within the first 15s and the reaction rate of the reference curve at the corresponding time point is compared, and when the difference at one time point is greater than or equal to 12%, it is determined that the sample has hook effect.

[0270] Or the average value of the reaction rate within the first 15s of the reaction is compared with the average value of the reference curve within the corresponding period, and when the difference is greater than or equal to 12%, it is determined that the sample has a hook effect.

[0271] In addition, according to Figure 9 , the hook effect can be determined by the profile of the reaction rate curve and the reference reaction rate curve. From Figure 9 It can be seen that in any one of samples 1-5, the reaction rate at the later time is always less than the reaction rate at the earlier time within a certain period of time, while the reference curve has a maximum value of the reaction rate within the period of time.

[0272] Alternatively, in some other cases where the alarm accuracy requirement is not high, the original reaction rate curve can also be used for alarm, for example, starting from 0s, the reaction rate of the next second and the previous second is compared continuously, and if the reaction rate is continuously decreasing over time within a period of time, it can be basically determined that the sample has a hook effect.

[0273] Example 3

[0274] The original reaction curve and the reference reaction curve obtained in Example 1 are respectively subjected to second-order derivation to obtain the original reaction acceleration curve and the reference reaction acceleration curve (see Figure 10 ).

[0275] According to the original reaction acceleration a1 and a2 at T1=5s and T2=20s after the second-order derivation of the original measurement curve, and the average value calculated according to the acceleration value of each second within 5-20s (see Table 7 below).

[0276] According to the reference reaction acceleration a 1(f) and a 2(f) at T1 and T2 after the second-order derivation of the reference reaction curve, and the average value calculated according to the acceleration value of each second within 5-20s (see Table 8 below).

[0277] Table 7: Original reaction acceleration and average reaction acceleration of SAA in the sample at T1 and T2

[0278]

[0279] Table 8: Reaction acceleration and average reaction acceleration of the reference reaction acceleration curve at T1 and T2

[0280]

[0281] The difference degree of the reaction acceleration at T1 and T2 is calculated according to the following formula:

[0282] δai % = | (a i - a i(f) ) / a i(f) | x 100%

[0283] Ti = 5s or 20s, when one of T1 and T2 exists i % ≥ 80% is determined to have a hook effect. Similarly, other time between T1 and T2 can be chosen, for example, 10s or 15s i % is greater than a threshold value to alarm. In addition, in this embodiment, since the original reaction acceleration curve and the reference reaction acceleration curve have time points close to each other or even cross each other, those skilled in the art should understand that it is best to avoid choosing these time points to calculate i %. Different time points can have different thresholds, and suitable thresholds can be obtained according to the statistics of known samples.

[0284] And calculate the difference of the average of 5s-20s according to the following formula:

[0285]

[0286] When Δa% ≥ 80%, it is determined to have a hook effect.

[0287] The specific results are shown in Table 9.

[0288] Table 9: Difference of reaction acceleration at 0s, 15s and average reaction acceleration at 5-20s

[0289]

[0290]

[0291] As can be seen from the above Table 9, the sample reaction acceleration has a large difference with the reference reaction acceleration within the first 15s. The difference of the reaction acceleration at a certain time within the first 15s and the reaction acceleration at the corresponding time of the reference curve is compared, and when the difference is greater than or equal to 80%, it is determined that the sample has a hook effect.

[0292] Or the average of the reaction acceleration within the first 15s and the average of the reference curve at the corresponding time are compared, and when the difference is greater than or equal to 80%, it is determined that the sample has a hook effect.

[0293] In addition, according to Figure 10 , the hook effect can be judged by the profile of the original reaction acceleration curve and the reference reaction acceleration curve. From Figure 10It can be seen that in any one of the samples 1-5, the reaction acceleration at the later time on the actual reaction acceleration curve is always greater than the reaction acceleration at the earlier time, and the reaction acceleration at the later time on the reference curve is always less than the reaction acceleration at the earlier time. Thus, it is judged that the sample has a hook effect.

[0294] Alternatively, when the alarm accuracy requirement is not high, the original reaction acceleration curve can be used for alarm. For example, starting from 0 seconds, the reaction acceleration of the next second and the previous second is continuously compared, and if the reaction acceleration is continuously increased over time within a period of time, it can be judged that the sample has a hook effect.

[0295] The above only describes the preferred embodiments of the present application, and does not limit the patent scope of the present application. Any equivalent structural transformation, direct / indirect application in other related technical fields, or the like within the inventive concept of the present application and using the content of the present application specification and drawings are included in the patent protection scope of the present application.

Claims

1. A method for identifying the hook effect in immunoturbidimetry, the method comprising: a) After the sample and reaction reagents are mixed to initiate the reaction of the analyte in the sample, the time intervals from T1 to T2 after the reaction begins are obtained. n T within the scheduled time period i Measurement value A at time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n To generate the response curve for the predetermined time period; b) Estimate the concentration C of the analyte based on the reaction curve over the predetermined time period. e ; c) Based on the estimated concentration C of the analyte e Obtain a pre-stored reference curve for this concentration; d) Compare the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period to determine whether the sample has a hook effect.

2. The method according to claim 1, wherein, In step a), T n The time T when the reaction has proceeded to 90% of its total reaction time is less than or equal to the time T. 90 .

3. The method according to claim 2, wherein, In step a), T n The time T when the reaction has proceeded to 70% of its total reaction time is less than or equal to the time T. 70 .

4. The method according to claim 2, wherein, In step a), T n The time T when the reaction has proceeded to 50% of its total reaction time is less than or equal to the time when the reaction has proceeded to 50% of its total reaction time. 50 .

5. The method according to claim 1, wherein, In step a), T n The time point at which the slope trend of the reference curve changes (less than or equal to the reference curve).

6. The method according to claim 1, wherein, The predetermined time period accounts for 10% to 70% of the total reaction time.

7. The method according to claim 6, wherein, The predetermined time period accounts for 15% to 50% of the total reaction time.

8. The method according to claim 6, wherein, The predetermined time period accounts for 15% to 40% of the total reaction time.

9. The method according to claim 1, wherein, The predetermined time period for generating the reaction curve of the analyte is from the start of the reaction until the reaction has proceeded to 10-70%.

10. The method according to claim 9, wherein, The predetermined time period for generating the reaction curve of the analyte is from the start of the reaction until the reaction has proceeded to 10-50%.

11. The method according to claim 9, wherein, The predetermined time period for generating the reaction curve of the analyte is from the start of the reaction until the reaction has proceeded to 10-40%.

12. The method according to any one of claims 1 to 11, wherein, The distribution information is selected from at least one of the measured value characteristics, reaction rate characteristics, and reaction acceleration characteristics.

13. The method according to any one of claims 1 to 11, wherein, If the difference between the distribution information of the reaction curve and the distribution information of the reference curve meets the predetermined characteristics, it is identified that the sample has a hook effect, and the detection is terminated.

14. The method according to any one of claims 1 to 11, wherein, The step of comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period includes: comparing the measured value of the reaction curve for the predetermined time period with the measured value of the reference curve for the corresponding time period.

15. The method according to any one of claims 1 to 11, wherein, The steps for comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period include: taking the first derivative of the reaction curve and the reference curve respectively to obtain the reaction rate curve and the reference rate curve, and then comparing them.

16. The method according to any one of claims 1 to 11, wherein, The steps for comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period include: taking the second derivative of the reaction curve and the reference curve respectively to obtain the reaction acceleration curve and the reference acceleration curve, and then comparing them.

17. The method according to any one of claims 1 to 11, wherein, The steps of comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period include: comparing the values ​​of the corresponding distribution information of the reaction curve and the reference curve at the same reaction time point, and / or comparing the average value of multiple corresponding distribution information of the reaction curve and the reference curve within the same time period.

18. The method according to any one of claims 1 to 11, wherein, In step d), the reaction curve is plotted on T... i Measurement value A at time i Reference value A at the corresponding time of the reference curve i(f) The response difference δ is calculated using the following formula. i %: d i %=[(A i –A i(f) ) / A i(f) ]×100% When δ i When the percentage is greater than or equal to a predetermined threshold δ1, the sample is determined to have a hook effect. or, The reaction curve was measured between T1 and T2. n The average measured value ā over the time period and the average reference value ā over the corresponding time period of the reference curve ā (f) The average response difference Δ% over this time period is calculated using the following formula: Δ%=[(a-a (f) ) / a (f) ]×100% When Δ% is greater than or equal to a predetermined threshold Δ1, the sample is determined to have a hook effect.

19. The method according to any one of claims 1 to 11, wherein, In step d), the reaction curve is plotted on T... i Measurement value A at time i Reference value A at the corresponding time of the reference curve i(f) The response difference δ is calculated using the following formula. i %: d i %=[(A i –A i(f) ) / A i(f) ]×100% When δ i When % is greater than or equal to the predetermined threshold δ2, and The reaction curve was measured between T1 and T2. n The average measured value ā over the time period and the average reference value ā over the corresponding time period of the reference curve ā (f) The average response difference Δ% over this time period is calculated using the following formula: Δ%=[(a-a (f) ) / a (f) ]×100% When Δ% is greater than or equal to a predetermined threshold Δ2, the sample is determined to have a hook effect.

20. The method according to any one of claims 1 to 11, wherein, Step d) includes: For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) Taking the first derivative with respect to time, we obtain the curves of reaction rate changing with time: v = A' = f'(T) and v' = A (f) '=f'(T (f) );as well as In T i The reaction rate value v at time i Compared with the reference reaction rate value v at the corresponding time i(f) The reaction rate difference δv is calculated using the following formula. i %: δv i %=(|v i -v i(f) | / v i(f) )×100% When δv i When % is greater than or equal to a predetermined threshold δv1, the sample is determined to have a hook effect. or, In T1~T n Average reaction rate over the time period Compared with the average reference reaction rate value within the corresponding time period The average reaction rate difference Δv% over this time period is calculated using the following formula: When Δv% is greater than or equal to a predetermined threshold Δv1, the sample is determined to have a hook effect.

21. The method according to any one of claims 1 to 11, wherein, Step d) includes: For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) Taking the first derivative with respect to time, we obtain the curves of reaction rate changing with time: v = A' = f'(T) and v' = A (f) '=f'(T (f) );as well as In T i The reaction rate value v at time i Compared with the reference reaction rate value v at the corresponding time i(f) The reaction rate difference δv is calculated using the following formula. i %: δv i %=(|v i -v i(f) | / v i(f) )×100% When δv i When % is greater than or equal to the predetermined threshold δv2, and In T1~T n Average reaction rate over the time period Compared with the average reference reaction rate value within the corresponding time period The average reaction rate difference Δv% over this time period is calculated using the following formula: When Δv% is greater than or equal to a predetermined threshold Δv2, the sample is determined to have a hook effect.

22. The method according to claim 20, wherein, T n Less than or equal to the difference δv i The time corresponding to when % is zero.

23. The method according to claim 21, wherein, T n Less than or equal to the difference δv i The time corresponding to when % is zero.

24. The method according to any one of claims 1 to 11, wherein, Step d) includes: For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) Taking the first derivative with respect to time, we obtain the curves of reaction rate changing with time: v = A' = f'(T) and v' = A (f) '=f'(T (f) );as well as When the conditions in T1 to T are simultaneously met n The reaction rate curve v = A' = f'(T) over the given time period shows an overall trend of decreasing reaction rate with time, and the reference reaction rate curve v' = A'(T) is used within the corresponding time period. (f) '=f'(T (f) When the reference reaction rate shows an overall trend of first increasing and then decreasing over time, the sample is determined to have a hook effect.

25. The method according to any one of claims 1 to 11, wherein, Step d) includes: For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) By performing second-order differentiation, we obtain the curves of reaction acceleration as a function of time: a = A” = f (T) and a f =A (f) =f(T) (f) );and In T i The reaction acceleration value a at time i Compared with the reference reaction acceleration value a at the corresponding time i(f) The reaction acceleration difference δa is calculated using the following formula. i %: δa i %=|(a i -in i(f) ) / in i(f) |×100% When δa i When % is greater than or equal to a predetermined threshold δa1, the sample is determined to have a hook effect. or, In T1~T n The average reaction acceleration value over the time period Compared with the average reference average reaction acceleration value within the corresponding time period The difference in average reaction acceleration Δa% over this time period is calculated using the following formula: When Δa% is greater than or equal to the predetermined threshold Δa1, the sample is determined to have a hook effect.

26. The method according to any one of claims 1 to 11, wherein, Step d) includes: For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) By performing second-order differentiation, we obtain the curves of reaction acceleration as a function of time: a = A” = f (T) and a (f) =A (f) =f(T) (f) );and In T i The reaction acceleration value a at time i Compared with the reference reaction acceleration value a at the corresponding time i(f) The reaction acceleration difference δa is calculated using the following formula. i %: δa i %=|(a i -in i(f) ) / in i(f) |×100% When δa i When % is greater than or equal to the predetermined threshold δa2, and In T1~T n The average reaction acceleration value over the time period Compared with the average reference average reaction acceleration value within the corresponding time period The difference in average reaction acceleration Δa% over this time period is calculated using the following formula: When Δa% is greater than or equal to the predetermined threshold Δa2, the sample is determined to have a hook effect.

27. The method according to claim 25, wherein, T n The value a is less than or equal to the reference reaction acceleration, which reaches a essentially constant value. const(f) The moment.

28. The method according to claim 26, wherein, T n The value a is less than or equal to the reference reaction acceleration, which reaches a essentially constant value. const(f) The moment.

29. The method according to any one of claims 1 to 11, wherein, Step d) includes: For the reaction curve A = f(T) and the reference curve A (f) =f(T) (f) By performing second-order differentiation, we obtain the curves of reaction acceleration as a function of time: a = A” = f (T) and a (f) =A (f) =f(T) (f) );and When the conditions in T1 to T are simultaneously met n The reaction acceleration curve a = A” = f (T) over the time period shows an overall trend of increasing reaction acceleration with time, and the reference reaction acceleration curve a (f) =A (f) =f(T) (f) When the overall trend of the reference reaction acceleration decreasing over time is observed, the sample is determined to have a hook effect.

30. A method for identifying the hook effect in immunoturbidimetry, the method comprising: a) After the sample and reaction reagents are mixed to initiate the reaction of the analyte in the sample, the time intervals from T1 to T2 after the reaction begins are obtained. n T within the scheduled time period i Acquire measurement value A at any time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n To generate the response curve for the predetermined time period; b) Differentiate the reaction curve A = f(T) over the predetermined time period with respect to time T, and determine whether the sample has a hook effect based on the profile of the differentiated curve.

31. The method according to claim 30, wherein, In step a), T n The time T when the reaction has proceeded to 90% of its total reaction time is less than or equal to the time T. 90 .

32. The method according to claim 31, wherein, In step a), T n The time T when the reaction has proceeded to 70% of its total reaction time is less than or equal to the time T. 70 .

33. The method according to claim 31, wherein, In step a), T n The time T when the reaction has proceeded to 50% of its total reaction time is less than or equal to the time when the reaction has proceeded to 50% of its total reaction time. 50 .

34. The method according to claim 30, wherein, The predetermined time period accounts for 10% to 70% of the total reaction time.

35. The method according to claim 34, wherein, The predetermined time period accounts for 15% to 50% of the total reaction time.

36. The method according to claim 34, wherein, The predetermined time period accounts for 15% to 40% of the total reaction time.

37. The method of claim 30, wherein, The time period used to generate the reaction curve of the analyte is from the start of the reaction until the reaction has proceeded to 10-70%.

38. The method according to claim 37, wherein, The time period used to generate the reaction curve of the analyte is from the start of the reaction until the reaction has proceeded to 10-50%.

39. The method according to claim 37, wherein, The time period used to generate the reaction curve of the analyte is from the start of the reaction until the reaction has progressed to 10-40%.

40. The method of claim 30, wherein, The differentiation is either first-order or second-order.

41. The method according to any one of claims 30 to 39, wherein, In step b), the first derivative of the reaction curve A = f(T) with respect to time T is taken to obtain the curve v = A' = f'(T) showing the change in reaction rate with time. When the condition is satisfied in T1~T n If the reaction rate curve v=A'=f'(T) within a certain time period shows an overall trend of decreasing reaction rate over time, then the sample is determined to have a hook effect.

42. The method according to any one of claims 30 to 39, wherein, In step b), the second derivative of the reaction curve A = f(T) with respect to time T is taken to obtain the curve a = A” = f(T) showing the change of reaction acceleration with time. When the condition is satisfied in T1~T n If the reaction acceleration curve a=A”=f(T) shows an overall trend of increasing reaction acceleration over time within a certain time period, it is determined that the sample has a hook effect.

43. The method according to claim 1 or 30, wherein, The sample was a whole blood sample from a mammal.

44. The method according to claim 43, wherein, The sample is a human whole blood sample.

45. The method according to claim 1 or 30, wherein, The analyte in the sample was protein.

46. ​​The method according to claim 45, wherein, The analyte in the sample is either C-reactive protein or serum amyloid protein.

47. The method according to claim 1 or 30, wherein, The method further includes, before mixing the sample with the reaction reagent, the steps of aspirating the sample into a reaction chamber, adding the reaction reagent and mixing, wherein the sample reacts with the reagent and the red blood cells in the sample are lysed.

48. The method according to claim 47, wherein, The reaction reagents include substances that can specifically react with the analyte.

49. The method according to claim 48, wherein, The reaction reagent includes a latex coated with the specific reactant, which can specifically react with the analyte.

50. The method according to claim 1 or 30, wherein, Before mixing the sample with the reaction reagent, the sample is divided into at least two equal parts, one part of which is used for routine blood tests, and the other part is used for analyte testing and subjected to the steps defined in claim 1 or 30.

51. A blood analysis system, the blood analysis system comprising: A sampling unit is used to acquire blood samples and transport the blood samples to a reaction unit; A reagent supply unit is used to store the first reaction reagent and supply it to the reaction unit as needed; The reaction section includes a first reaction chamber for mixing the blood sample with the first reaction reagent to prepare a first test solution; The detection system includes a light source and a detector for detecting the first test solution, used to acquire the measured value of the test solution; A controller is coupled to the sampling unit, reagent supply unit, reaction unit, and detection system, and controls the operation of the sampling unit, reagent supply unit, reaction unit, and detection system; and The processor, which is coupled to the detection system, The processor receives the reaction start time T1~T1 from the detection system. n T within the scheduled time period i Measurement value A at time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n The reaction curve for the predetermined time period is generated; the concentration C of the analyte is estimated based on the reaction curve for the predetermined time period and a pre-stored calibration curve. e ; Based on the estimated concentration C of the analyte e Obtain a pre-stored reference curve for this concentration; The distribution information of the reaction curve for the predetermined time period is compared with the distribution information of the reference curve for the corresponding time period to determine whether the blood sample exhibits a hook effect. Alternatively, the processor obtains the reaction start time T1 to T2 from the detection system. n T within the scheduled time period i Acquire measurement value A at any time i Where i is an integer from 1 to n, thus obtaining multiple measurement values ​​A1 to A2. n The processor generates a reaction curve for the predetermined time period; it differentiates the reaction curve A = f(T) within the predetermined time period with respect to time T, and determines whether the blood sample has a hook effect based on the curve profile after differentiation. The processor then outputs the determination result to the controller.

52. The blood analysis system according to claim 51, wherein, The step of the processor comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period includes: comparing the measured value of the reaction curve for the predetermined time period with the measured value of the reference curve for the corresponding time period.

53. The blood analysis system according to claim 51, wherein, The step of the processor comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period includes: performing first-order derivatives on the reaction curve and the reference curve respectively to obtain the reaction rate curve and the reference rate curve, and then comparing them.

54. The blood analysis system according to claim 51, wherein, The step of the processor comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period includes: performing second derivatives on the reaction curve and the reference curve to obtain the reaction acceleration curve and the reference acceleration curve, and then comparing them.

55. The blood analysis system according to any one of claims 51 to 54, wherein, The step of the processor comparing the distribution information of the reaction curve for the predetermined time period with the distribution information of the reference curve for the corresponding time period includes: comparing the values ​​of the corresponding distribution information of the reaction curve and the reference curve at the same reaction time point, and / or comparing the average value of multiple corresponding distribution information of the reaction curve and the reference curve within the same time period.

56. The blood analysis system according to any one of claims 51 to 54, wherein the controller, upon receiving a result indicating the presence of a hook effect in the current test sample, controls the testing system to stop testing; The sampling unit is controlled to resample the blood sample and transport the sample to the first reaction chamber of the reaction unit; The reagent supply unit is controlled to supply the first reaction reagent to the first reaction chamber to prepare a second test solution, wherein the dilution factor of the sample in the second test solution is greater than the dilution factor of the sample in the first test solution; and The detection system is controlled to detect the second test solution.

57. The blood analysis system according to any one of claims 51 to 54, wherein, The detector includes a photometer.

58. The blood analysis system according to claim 57, wherein, The detectors include turbidimeters and / or turbidimeters.

59. The blood analysis system according to any one of claims 51 to 54, wherein, The blood sample is a whole blood sample, and the first reaction reagent includes a hemolysin for lysing red blood cells in the sample and a latex reagent for performing an immunoturbidimetric reaction with the analyte in the blood sample.

60. The blood analysis system according to any one of claims 51 to 54, further comprising a second detection system, wherein the reaction unit further comprises a second reaction chamber; the second detection system comprises a light source, a flow chamber for cells to pass through in sequence, a liquid path system, and a second detector; The controller controls the sampling unit to divide the sample into two parts, which are then transported to the first reaction chamber and the second reaction chamber, respectively; it controls the reagent supply unit to transport the second reagent to the second reaction chamber, where the sample reacts with the second reagent to obtain a third test solution; and it controls the second detection system, under the drive of the liquid circuit system, to transport the third test solution to the flow chamber, where the light source illuminates the flow chamber, and the second detector collects the scattered light signal generated by the cells. The processor obtains the scattered light signal and classifies the white blood cells in the sample into at least three categories: lymphocytes, monocytes, and neutrophils based on the scattered light signal.

61. The blood analysis system according to claim 60, wherein, The second detection system further includes a third detector, which collects fluorescence signals generated by cells and classifies white blood cells into at least four categories: lymphocytes, monocytes, neutrophils, and eosinophils based on the scattered light signal and fluorescence signal.

62. The blood analysis system according to claim 60, wherein, The second reagent includes a hemolytic agent and a staining agent.

63. The blood analysis system according to claim 61, wherein, The second reagent includes a hemolytic agent and a staining agent.

64. A computer-readable medium storing executable instructions, wherein, The computer-readable medium is configured to cause a processor to execute the executable instructions to implement the hook effect identification method for immunoturbidimetry as described in any one of claims 1 to 42.

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