WO2020103051A1

WO2020103051A1 - Method for measuring sample absorbance difference, sample analyzer and storage medium

Info

Publication number: WO2020103051A1
Application number: PCT/CN2018/116778
Authority: WO
Inventors: 李聪; 郭文恒; 李坷坷
Original assignee: 北京普利生仪器有限公司
Priority date: 2018-11-21
Filing date: 2018-11-21
Publication date: 2020-05-28
Also published as: CN112912712A; CN112912712B

Abstract

A method for measuring a sample absorbance difference, a sample analyzer and a storage medium. The method comprises the following steps: acquiring absorbance data of a sample object; according to the absorbance data, using a two-point assay to calculate a first absorbance difference, and according to the absorbance data, using a rate assay to calculate a second absorbance difference; substituting the first absorbance difference and the second absorbance difference into a pre-determined weighted average function used to calculate a final absorbance difference; according to a pre-determined function relation, determining a weighting of the first absorbance difference and a weighting of the second absorbance difference which satisfy the weighted average function, and a final absorbance difference. There is also a sample analyzer, comprising a light source (132), a detector (134) and a processor (136).

Description

Measuring method of sample absorbance difference, sample analyzer and storage medium

Technical field

The present application relates to the field of optical measurement, and in particular, to a method for measuring a difference in absorbance of a sample, a sample analyzer, and a storage medium.

Background technique

In the prior art, the automatic coagulation analyzer generally uses immunoturbidimetry to test the concentration of D-dimer (DD), fibrin (pro) degradation products (FDP) and other items, and the most basic biochemical reaction calculation method has three Species: endpoint method (1-Point), 2-point method (2-Point) and rate method (Rate). Among them, the endpoint method is: the instrument only detects the absorbance value at a certain time point of the biochemical reaction, which is susceptible to interference, and is rarely used in automatic coagulation analyzers; the two-point method is: the instrument detects the absorbance value at two time points of the biochemical reaction , The absorbance value at the second time point minus the absorbance value at the first time point to obtain the difference in absorbance (absorbance difference). The two-point method is only suitable for the measurement of the linear period of the reaction rate, but as the substrate is continuously consumed, the entire reaction rate is continuously reduced, so the two-point method has a maximum limit that can be measured, and absorbance difference saturation will occur after the limit is exceeded , Or even lower, thereby limiting the linear ability of the high value part. Rate method: The instrument continuously monitors the change in absorbance caused by changes in the substrate or product content during the biochemical reaction, obtains the rate of change in absorbance, and determines the difference in absorbance based on the rate of change in absorbance. The key of the rate method is to accurately describe the relationship between the reaction rate and time. The signal-to-noise ratio is relatively low during the detection of low-value samples. The rate of continuous monitoring is easily interfered, and it is difficult to accurately reflect the biochemical reaction process, resulting in low value repeatability bad.

At present, most automatic coagulation analyzers support the two-point method and the rate method. However, no matter what calculation method is used, they cannot meet the low value repeatability and high value linearity at the same time.

In response to the above problems, no effective solution has been proposed yet.

Summary of the invention

The embodiments of the present application provide a method for measuring a sample absorbance difference, a sample analyzer, and a storage medium, to at least solve the problem of measuring repeatability when measuring absorbance difference, when the absorbance difference is low and the absorbance difference is high. Technical issues with linear measurement range.

According to an aspect of an embodiment of the present application, there is provided a method for measuring a sample absorbance difference, including; acquiring absorbance data of a sample object; calculating the first absorbance difference using the two-point method according to the absorbance data, and calculating the Second absorbance difference; substitute the first absorbance difference and the second absorbance difference into the preset weighted average function used to calculate the final absorbance difference; determine the weight and the second absorbance of the first absorbance difference satisfying the weighted average function according to the preset functional relationship The weight of the difference and the difference in the final absorbance, wherein the preset functional relationship is a function relationship for the weight of the first difference in absorbance, the weight of the second difference in absorbance and the difference in absorbance.

Optionally, after determining the weight of the first absorbance difference and the weight of the second absorbance difference and the final absorbance difference satisfying the weighted average function according to the preset function relationship, the method further includes: determining the corresponding to the final absorbance difference according to the preset correspondence Sample concentration, where the preset correspondence is the correspondence between absorbance difference and sample concentration.

Optionally, the preset functional relationship includes: within the set definition domain, if the absorbance difference is less than the set threshold, the weight of the first absorbance difference is greater than the weight of the second absorbance difference; if the absorbance difference is greater than the set threshold, the first The weight of one difference in absorbance is less than the weight of the difference in second absorbance.

Optionally, if the difference in absorbance is less than the set threshold, the absolute value of the difference between the weight of the first absorbance difference and the weight of the difference in the second absorbance is negatively related to the difference in absorbance; if the difference in absorbance is greater than the set threshold, the absolute value of the difference in weight Positively correlated with the difference in absorbance.

Optionally, if the difference in absorbance is less than the set threshold, the absolute value of the weighted phase difference decreases as the difference in absorbance increases, and the rate of decrease in the absolute value of the weighted phase difference is positively correlated with the difference in absorbance.

Alternatively, if the absorbance difference is greater than the set threshold, the absolute value of the weighted phase difference increases as the absorbance difference increases, and the rate of increase in the absolute value of the weighted phase difference is inversely related to the absorbance difference.

Optionally, the set threshold is set by the user.

Optionally, the speed of change of the absolute value of the weight difference is set by the user.

Optionally, the preset functional relationship further includes: within the set definition domain, if the absorbance difference is equal to the set threshold, the weight of the first absorbance difference and the second absorbance difference are the same.

Optionally, the weighted average function is: F (x) = a · x + b · (cx) + d; where a is the second absorbance difference, x is the weight of the second absorbance difference, and b is the first absorbance difference , (Cx) is the weight of the first absorbance difference, c is a constant greater than or equal to x, d is a constant greater than or equal to 0, and F (x) is the final absorbance difference.

Optionally, the method is applied to a sample analyzer.

According to an aspect of an embodiment of the present application, there is provided a sample analyzer, including: a light source, a detector, and a processor; wherein, the light source is used to emit a light beam for irradiating the sample; the detector is used to detect the light beam after the sample is irradiated Luminous flux data; the processor runs the program, where the program executes the following processing steps on the data output from the detector when the program is running: calculating the absorbance data based on the luminous flux data; calculating the first difference in absorbance based on the absorbance data using the two-point method, and applying the absorbance data Rate method to calculate the second absorbance difference; substitute the first absorbance difference and the second absorbance difference into the preset weighted average function used to calculate the final absorbance difference; determine the weight of the first absorbance difference satisfying the weighted average function according to the preset function relationship And the weight of the second absorbance difference and the final absorbance difference, where the preset functional relationship is a function relationship for the weight of the first absorbance difference, the weight of the second absorbance difference, and the absorbance difference.

Optionally, the processor is further configured to: determine the sample concentration corresponding to the final absorbance difference according to a preset correspondence, where the preset correspondence is the correspondence between the absorbance difference and the sample concentration.

Optionally, if the difference in absorbance is less than the set threshold, the absolute value of the difference between the weight of the first absorbance difference and the weight of the difference in the second absorbance is negatively related to the difference in absorbance; Positively correlated with the difference in absorbance.

Optionally, the set threshold is set by the user.

Optionally, the weighted average function is: F (x) = a · x + b · (cx) + d; where a is the second absorbance difference, x is the weight of the second absorbance difference, and b is the first absorbance difference , Cx is the weight of the first difference in absorbance, c is a constant greater than or equal to x, d is a constant greater than or equal to 0, and F (x) is the final difference in absorbance.

According to an aspect of an embodiment of the present application, one or more non-volatile computer-readable storage media are provided on which a computer program is stored, and when the computer program is executed by a processor, the above-mentioned method for measuring the difference in absorbance of a sample is realized .

In the embodiment of the present application, the two-point method is used to calculate the first absorbance difference according to the absorbance data of the sample object, and the second absorbance difference is calculated using the rate method according to the absorbance data; and the first absorbance satisfying the weighted average function is determined according to the preset function relationship The weight of the difference, the weight of the second absorbance difference, and the final absorbance difference, where the preset functional relationship is a function relationship for the weight of the first absorbance difference, the weight of the second absorbance difference, and the absorbance difference. The two-point method is adopted by the low value part to obtain better low value repeatability; the rate method is used to obtain the monotonicity of the calculated value of the difference in absorbance and the higher linear measurement range; the low value and the high value Partially well-connected, there is no technical effect of faults or jumps; thus solving the technical problem of measuring the repeatability of the measurement when the absorbance difference is low, and the linear measurement range when the absorbance difference is high.

BRIEF DESCRIPTION

The drawings described herein are used to provide a further understanding of the present application and form a part of the present application. The schematic embodiments and descriptions of the present application are used to explain the present application and do not constitute an undue limitation on the present application. In the drawings:

FIG. 1a is a schematic structural diagram of a sample analyzer according to an embodiment of the present application;

FIG. 1b is a schematic flowchart of an optional method for measuring absorbance of a sample according to an embodiment of the present application;

FIG. 2 is a flowchart of calculating the first absorbance difference by the two-point method provided in the embodiment of the present application;

FIG. 3 is a flowchart of calculating the second difference in absorbance by the rate method provided by the embodiment of the present application;

4 is a flow chart of another rate method for calculating a second difference in absorbance provided by an embodiment of the present application;

FIG. 5 is a schematic diagram of weight changes of the two-point method and the rate method determined when the preset function is based on a hyperbolic tangent function according to an embodiment of the present application; FIG.

6 is a schematic diagram of the weight change of the two-point method and the rate method determined when the preset function is based on a first-order system according to an embodiment of the present application;

7 is a schematic diagram of a change in the weight of the first absorbance difference and the second absorbance difference determined by the preset function according to an embodiment of the present application;

8 is a schematic flowchart of a method for measuring a difference in absorbance of a sample provided by an embodiment of the present application;

9 is a schematic flowchart of a method for measuring a difference in absorbance of a sample according to another embodiment of the present application;

Figures 10a-10d are schematic diagrams of the repetitive calculation results of the two-point method and the rate method for calculating the low value part of the difference in absorbance;

11 is a schematic diagram of comparison between the two-point method, the rate method, and the absorbance difference calculated using the measurement method of the sample absorbance difference described in this application when the absorbance difference is a high value;

12 is a flowchart of a working method corresponding to a sample analyzer provided by an embodiment of the present application.

detailed description

In order to enable those skilled in the art to better understand the solutions of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only It is a part of the embodiments of this application, but not all the embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the scope of protection of this application.

It should be noted that the terms “first” and “second” in the description and claims of the present application and the above drawings are used to distinguish similar objects, and do not have to be used to describe a specific order or sequence. It should be understood that the data used in this way can be interchanged under appropriate circumstances, so that the embodiments of the present application described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, processes, methods, systems, products or devices that contain a series of steps or units need not be limited to those clearly listed Those steps or units, but may include other steps or units not explicitly listed or inherent to these processes, methods, products, or equipment.

In order to better understand the embodiments of the present application, the following briefly describes the terms involved in the embodiments of the present application as follows:

Rate method: Also known as dynamic method and kinetic method, the instrument continuously monitors the absorbance change caused by the change of the substrate or product content during the biochemical reaction, obtains the rate of change in absorbance, and finds the moment when the reaction rate is maximum within the specified sampling time. It is extended to both sides with the center as the center to find an interval that approximately meets the linear reaction rate, and the absorbance difference in this interval is calculated as the absorbance difference of the entire reaction process.

The key of the rate method is to accurately describe the relationship between the reaction rate and time, and then accurately determine the rate of change of absorbance.

Two-point method: The absorbance values at two time points of the biochemical reaction are detected by the instrument, which are the sampling start point and the sampling end point, respectively. The absorbance difference of the method is obtained by subtracting the absorbance of the sampling start point from the absorbance of the sampling end point. The two-point method is only suitable for the determination of the linear phase of the reaction rate. It cannot accurately describe the process that the entire reaction rate is continuously decreasing as the substrate is continuously consumed.

The main difference between the above two-point method and the rate method in describing the reaction process is that the two-point method is based on that the reaction rate does not change with the consumption of the substrate (expressed as the test time), and the rate method is based on the reaction rate. Changes with consumption.

FIG. 1a is a schematic structural diagram of a sample analyzer according to an embodiment of the present application. As shown in FIG. 1a, the sample analysis includes: a light source 132, a detector 134, and a processor 136, wherein the light source 132 is used for emitting Sample beam;

The detector 134 is used to detect the luminous flux data generated after the beam illuminates the sample;

The processor 136 runs a program, where the following steps are performed on the data output from the detector when the program is running: calculating the absorbance data based on the luminous flux data; calculating the first absorbance difference using the two-point method based on the absorbance data, and calculating the first absorbance data based on the absorbance data using the rate method Second absorbance difference; substitute the first absorbance difference and the second absorbance difference into the preset weighted average function used to calculate the final absorbance difference; determine the weight and the second absorbance of the first absorbance difference satisfying the weighted average function according to the preset functional relationship The weight of the difference and the final difference in absorbance; wherein, the preset functional relationship is a function of the weight for the first difference in absorbance, the weight of the second difference in absorbance, and the difference in absorbance.

The above-mentioned sample analyzers include, but are not limited to, blood analyzers, biochemical analyzers, immunoassay analyzers, coagulation analyzers, and other in vitro diagnostic equipment.

The embodiment of the present application provides a method for measuring the absorbance of a sample. The method runs on the above-mentioned sample analyzer. It should be noted that the steps shown in the flowchart of the following drawings can be executed in a computer system such as a set of computer-executable instructions, and although the logical sequence is shown in the flowchart, in some cases The steps shown or described can be performed in a different order than here.

FIG. 1b is a schematic flowchart of a method for measuring a sample absorbance according to an embodiment of the present application. As shown in FIG. 1b, the method includes at least steps S102-S108, in which:

Step S102, acquiring the absorbance data of the sample object;

In some optional embodiments of the present application, the absorbance data of the sample object may be obtained by performing the following steps S1022-S1024: Step S1022, obtaining the luminous flux data by an instrument that detects luminous flux; here, the luminous flux data includes transmitted light and / or scattering Luminous flux data of light; Step S1024: Determine absorbance data according to the aforementioned luminous flux data.

After acquiring the absorbance data, step S104 can be performed.

Step S104: Apply a two-point method to calculate the first difference in absorbance according to the absorbance data, and apply a rate method to calculate the second difference in absorbance according to the absorbance data;

In some optional embodiments of the present application, FIG. 2 is a flowchart of calculating the first absorbance difference by the two-point method provided in the embodiment of the present application; calculating the first absorbance difference by applying the two-point method according to the absorbance data can be performed by the following step S202 -S206 realization:

Step S202, obtaining a luminous flux curve; the luminous flux curve is a curve generated according to the luminous flux data;

In some optional embodiments of the present application, the luminous flux data may be sampled, and a luminous flux curve may be drawn according to the sampling result, where the luminous flux is the luminous flux of scattered light and / or transmitted light after the light beam emitted by the light source illuminates the sample;

Luminous flux data corresponds to time information; for example, luminous flux data can be expressed by the following function:

Intensity = f (time), where the above Intensity represents the luminous flux data of the scattered light and / or transmitted light after the beam emitted by the light source illuminates the sample, and f (time) represents the luminous flux data corresponding to the time information; it can be based on the luminous flux data and the time information Corresponding relationship, draw the luminous flux curve.

Step S204: Determine an absorbance curve according to the luminous flux curve; the absorbance curve is a curve generated according to the absorbance data;

In some optional embodiments of the present application, the absorbance data and the luminous flux data have the following relationship:

Abs = log10 (I0 / Intensity);

Where Abs represents absorbance data, I0 represents incident luminous flux data, reference luminous flux data, or other luminous flux data before the beam emitted by the light source illuminates the sample, based on the luminous flux of scattered light and / or transmitted light after the beam emitted by the light source illuminates the sample The data Intensity and the incident light flux data I0 before the beam emitted by the light source irradiates the sample determine the absorbance data or the absorbance curve.

Step S206: Determine the first difference in absorbance based on the absorbance curve.

In some alternative embodiments of the present application, the absorbance difference may be represented by the following _{formula: dOD Twopoint = Abs (EndPoint)} -Abs (StartPoint);

_Wherein, dOD Twopoint represents a first difference in absorbance, Abs (EndPoint) indicates the termination time point corresponding to the absorbance data, Abs (StartPoint) represents an initial time point absorbance data.

In some optional embodiments of the present application, FIG. 3 is a flowchart of calculating the second difference in absorbance by the rate method provided in the embodiment of the present application; applying the rate method to calculate the second difference in absorbance according to the absorbance data may be performed through the following steps S302-S310 achieve:

Step S302, collecting the luminous flux curve that changes with the reaction time;

Step S304, find the moment with the fastest reaction rate and record it as MaxPoint;

Step S306, find an interval approximately satisfying the linear reaction process near MaxPoint, and record the start and end points of the interval as Cut_Min and Cut_Max;

Step S308: Calculate the difference in absorbance between the start point and the end point of the interval, and record them as Abs (Cut_Min) and Abs (Cut_Max), respectively;

In step S310, the difference in absorbance is calculated: dOD _RateMethod = Abs (Cut_Max) -Abs (Cut_Min).

In some optional embodiments of the present application, FIG. 4 is a flowchart of another rate method for calculating a second absorbance difference provided by an embodiment of the present application; applying the rate method to calculate the second absorbance difference according to absorbance data may be performed by the following steps S402-S416 realize:

Step S402, the instrument collects the luminous flux curve: Intensity = f (time); wherein, the above Intensity represents the luminous flux data of the scattered light and / or transmitted light after the beam emitted by the light source illuminates the sample, and f (time) represents the luminous flux data and time information Correspondence; based on the correspondence between luminous flux data and time information, a luminous flux curve can be drawn;

Step S404, calculate and obtain the absorbance curve: Abs = log10 (I0 / Intensity); Abs represents the absorbance data, I0 represents the incident luminous flux data, reference luminous flux data, or other reference luminous flux data before the sample is irradiated by the light beam emitted by the light source, based on the light source The luminous flux data of scattered light and / or transmitted light after the sample beam illuminates the sample and the incident luminous flux data I0 before the sample beam from the light source determines the absorbance data or absorbance curve

Step S406, within the analysis interval, an N-order polynomial is fitted to the absorbance curve (N≥2) to obtain Abs_fit;

In step S408, the Abs_fit curve is derived, the tangent equation is obtained, and the MaxPoint time point is obtained;

Step S410, starting from MaxPoint and extending the linear range with the shortest regression time as a unit;

Step S412, the integral area between the original absorbance curve and the fastest point tangent is obtained, and the maximum linear range less than the set threshold is obtained;

Step S414, obtaining the tangent equation Abs_cut and the limit points of the linear range: Cut_Min and Cut_Max;

Step S416, calculating the difference in absorbance: dOD _RateMethod = Abs_cut (Cut_Max) -Abs_cut (Cut_Min);

After calculating the first difference in absorbance and the second difference in absorbance, step S106 can be performed.

Step S106, substituting the first absorbance difference and the second absorbance difference into a preset weighted average function (which may also be called a weight allocation function or a proportional allocation function) used to calculate the final absorbance difference;

In some optional embodiments of the present application, the weighted average function is F (x) = a · x + b · (cx) + d; where a is the second absorbance difference and x is the second absorbance difference Weight, b is the first difference in absorbance, cx is the weight of the first difference in absorbance, c is a constant greater than or equal to x (usually 1), d is a constant greater than or equal to 0 (usually 0), F (x) For the final absorbance difference, · indicates the multiplication sign. When c = 1 and d = 0, the weighted average function is F (x) = a · x + b · (1-x), that is, the sum of the weight of the first absorbance difference and the weight of the second absorbance difference is 1, Thus, if any weight is determined, another weight can be determined. In the following description, the weighted average function is mainly described as F (x) = a · x + b · (1-x).

Step S108: Determine the weight of the first absorbance difference, the weight of the second absorbance difference and the final absorbance difference satisfying the weighted average function according to the preset function relationship; wherein the preset function relationship is the weight for the first absorbance difference, the first Second, the weight of the difference in absorbance is a function of the difference in absorbance.

You can pre-store the functional relationship of "weight of the first absorbance difference-weight of the second absorbance difference-absorbance difference", such as pre-stored iterative equations or a large number of "weight of the first absorbance difference-weight of the second absorbance difference-absorbance Poor data combination, such as "50% -50% -4000", "98% -2% -2000", "2% -98% -6000" and so on. Then, in the weighted average function that has been substituted into the first absorbance difference and the second absorbance difference, iteratively search for the data combination satisfying the weighted average function in the database, so as to determine the weight of the first absorbance difference and the weight of the second absorbance difference and the final Poor absorbance. For example, after calculating the second absorbance difference a and the first absorbance difference b, substitute the weighted average function as F (x) = a · x + b · (1-x), and then iterate from the database to find the weighting The data combination of the average function.

In an alternative embodiment, the process of iteratively searching for the data combination in step S108 may be represented as the following processing, but not limited to this: determine the target absorbance difference according to the weight of the first absorbance difference and the weight of the second absorbance difference; determine the target Whether the weight of the difference in absorbance, the weight of the first difference in absorbance and the weight of the second difference in absorbance satisfy the preset weighted average function; when the judgment result is yes, it is determined that the target difference in absorbance is the final difference in absorbance.

In some optional embodiments of the present application, the preset function corresponding to the above preset function relationship is mainly for realizing: if the difference in absorbance is low, the result of the corresponding final absorbance difference is infinitely close to the first absorbance difference; absorbance If the difference is higher, the result of the corresponding final absorbance difference is infinitely closer to the second absorbance difference; if the absorbance difference is in the middle, the transition interval is as smooth as possible, without jumps, overlaps, and faults.

In some optional embodiments of the present application, the preset functional relationship also needs to be considered: within the set definition domain, if the absorbance difference is less than a set threshold, the weight of the first absorbance difference is greater than the first The weight of the second absorbance difference; if the absorbance difference is greater than the set threshold, the weight of the first absorbance difference is less than the weight of the second absorbance difference. In some embodiments, the set threshold can be set by the user.

Since any weight can be determined, another weight can be determined. Therefore, the preset function in the preset function relationship may be a function about one of the weights, for example, a function about the weight of the second difference in absorbance.

The preset function can be Prop = {tanh [(dOD-dOD0) * k + 1]} * 0.5; where tanh represents the hyperbolic tangent function, and the basic function of the hyperbolic tangent function is: tanh (x) = [exp ( -x) -exp (x)] / [exp (-x) + exp (x)]; where Prop is the weight of the second absorbance difference, dOD is the value of the absorbance difference, and dOD0 is the above-mentioned preset threshold, k For the decay time coefficient, * represents the multiplication sign; where, dOD0 and k can be set by the user. When dOD = dOD0, the weight of the first difference in absorbance and the weight of the second difference in absorbance are both 0.5. Referring to FIG. 5, dOD0 is 5000. When dOD = 5000, the weight of the first absorbance difference and the weight of the second absorbance difference are both 0.5. Therefore, in some embodiments, in the defined domain of the setting, if the difference in absorbance is equal to the set threshold, the weight of the first difference in absorbance and the weight of the second difference in absorbance are the same.

In some optional embodiments of the present application, if the difference in absorbance is less than the set threshold, the absolute value of the difference between the weight of the first difference in absorbance and the weight of the second difference in absorbance is negatively correlated with the difference in absorbance; If the difference in absorbance is greater than the set threshold, the absolute value of the weight difference is positively related to the difference in absorbance. Continuing to take the above preset function as an example, please refer to FIG. 5, which is a schematic diagram of the weight change of the two-point method and the rate method determined when the preset function is based on the hyperbolic tangent function provided by an embodiment of the present application; The axis is the absorbance difference, the vertical axis is the weight (proportion), dOD0 is 5000, when the absorbance difference dOD is less than 5000, as the dOD decreases, the weight of the first absorbance difference gradually increases and the weight of the second absorbance difference gradually decreases , Therefore, the absolute value of the difference between the weight of the first absorbance difference and the weight of the second absorbance difference becomes larger gradually, so the absolute value of the difference between the weight of the first absorbance difference and the weight of the second absorbance difference is negatively correlated with the absorbance difference . When dOD is greater than 5000, the weight of the first absorbance difference gradually becomes smaller and the weight of the second absorbance difference gradually becomes larger as dOD increases. Therefore, the weight of the first absorbance difference and the weight of the second absorbance difference are different. The absolute value of becomes gradually larger, so the weight difference between the weight of the first absorbance difference and the weight of the second absorbance difference is positively correlated with the absorbance difference.

In some optional embodiments of the present application, if the absorbance difference is less than the set threshold, the absolute value of the weighted phase difference decreases as the absorbance difference increases, and the absolute value of the weighted phase difference The reduction speed is positively correlated with the difference in absorbance. In some optional embodiments of the present application, if the absorbance difference is greater than the set threshold, the absolute value of the weighted phase difference increases as the absorbance difference increases, and the absolute value of the weighted phase difference The increase speed is inversely related to the difference in absorbance. Continuing to take the above preset function as an example, please refer to FIG. 5, dOD0 is 5000, when the absorbance difference dOD is less than 5000, as the dOD decreases, the weight of the first absorbance difference gradually increases but the rate of change decreases, while the second absorbance The weight of the difference gradually decreases but the rate of change also decreases. Therefore, as dOD decreases, the absolute value of the difference between the weight of the first absorbance difference and the weight of the second absorbance difference gradually increases but the rate of change is decreasing. That is, the rate of decrease in the absolute value of the weight difference is positively correlated with the difference in absorbance. When the absorbance difference dOD is greater than 5000, as the dOD increases, the weight of the first absorbance difference gradually decreases but the rate of change decreases, while the weight of the second absorbance difference gradually increases but the rate of change also decreases. Becomes larger, the absolute value of the weight difference between the weight of the first absorbance difference and the weight of the weight of the second absorbance difference gradually increases but the rate of change is decreasing, that is, the rate of increase in the absolute value of the weight difference is inversely related to the absorbance difference. In some optional embodiments of the present application, the speed of decreasing or increasing the absolute value of the weight difference is set by the user. For example, k in the preset function can be set by the user.

Of course, in other embodiments, the preset function may also be a first-order system, see FIG. 6, for example, y = 1-exp (-x / τ), where the horizontal axis is the absorbance difference and the vertical axis is the weight (proportion), τ is the response time, x is the absorbance difference dOD, and y is the weight of the second absorbance difference.

7 is a schematic diagram of changes in the weights of the first difference in absorbance and the second difference in absorbance determined by the preset function. In FIG. 7, the horizontal axis is the absorbance difference, and the vertical axis is the ratio of the weight of the first absorbance difference corresponding to the two-point method and the second absorbance difference corresponding to the rate method. In the low value part of the absorbance difference, the preset function can make the total absorbance difference infinitely close to the calculation result of the two-point method, and make full use of the linear fitting process of the two-point method to resist noise interference and improve the signal-to-noise ratio for better Low value repeatability; in the high value part of the absorbance difference, the preset function can make the total absorbance difference infinitely close to the calculation result of the rate method, make full use of the rate method to actively reflect the change of the reaction rate with time to obtain the difference in absorbance Monotonicity of calculated values and higher linear measurement range.

For example: when the threshold is set to 4000, at the position where dOD ≤ 1000 (the interval where the low value belongs), the proportion of the value calculated by the two-point method to the total absorbance difference is ≥99.75%, which inevitably determines the value of the total absorbance difference; at dOD ≥7000 The position (near the median value, which is not yet a high value part), the calculated value of the two-point method accounts for less than 0.25% of the total absorbance difference, the total absorbance difference is consistent with the rate method calculation results; the two-point method and the rate method are in the total absorbance The distribution ratio in the difference is continuously adjusted, the derivative is continuous, and there is no mutation, so the whole process is also continuous.

In some optional embodiments of the present application, a schematic flowchart of a method for measuring the difference in absorbance of a sample as shown in FIG. 8 is provided; the method includes the following steps S802, S8022-S8024, S8042-S8048, and S806 Where steps S8022-S8024 are used to calculate the first difference in absorbance, and steps S8042-step S8048 are used to calculate the second difference in absorbance.

Step S802, collecting the luminous flux curve that changes with the reaction time;

S8022-S8024 are as follows:

Step S8022: Calculate the absorbance of the sampling start point and the sampling end point, which are respectively recorded as: Abs (StartPoint), Abs (EndPoint);

Step S8024, a first calculated absorbance _{difference: dOD Twopointn = Abs (EndPoint)} -Abs (StartPoint);

Step S8042-step S8048 are as follows:

Step S8042: Find the moment with the fastest reaction rate and record it as MaxPoint;

Step S8044: Find an interval that satisfies the linear response process approximately near MaxPoint, and record the start and end points of the interval as Cut_Min and Cut_Max;

Step S8046: Calculate the difference in absorbance between the start point and the end point of the interval, and record them as Abs (Cut_Min) and Abs (Cut_Max) respectively;

Step S8048, the second difference in absorbance is calculated: dOD _RateMethod = Abs (Cut_Max) -Abs (Cut_Min);

After obtaining a first absorbance and the second absorbance difference _dOD Twopoint difference _dOD RateMethod Through the above steps, perform step S806, the calculated weight of the final absorbance difference that is a weighted average by assigning a weight function;

Step S806, the obtained according to a certain ratio of the final absorbance difference _{dOD = dOD Twopoint * prop (dOD} ) + dOD RateMethod * (1-prop (dOD));

Wherein the absorbance difference (first difference in _absorbance) dOD Twopoint is a two-point _calculation, dOD RateMethod absorbance difference (second difference in absorbance) was calculated by a rate method, prop (DOD) for the two-point method to calculate the absorbance difference is occupied Weight, 1-prop (dOD) is the weight of the difference in absorbance calculated by the rate method.

In some optional embodiments of the present application, a schematic flowchart of a method for measuring the difference in absorbance of a sample as shown in FIG. 9 is provided; the method includes the following steps S902-Step S904, Step S9062-Step S9064, Step S9066-Step S90616 and step S908, wherein step S9062-step S9064 are used to calculate the first absorbance difference, and S9066-step S90616 are used to calculate the second absorbance difference.

Steps S902-S904 are as follows:

Step S902, the instrument collects the luminous flux curve: Intensity = f (time);

Step S904, calculating and obtaining the absorbance curve: Abs = log10 (I0 / Intensity);

Step S9062-step S9064 are as follows:

Step S9062, within the analysis interval, linearly fit the absorbance curve: Abs _fit ;

Step S9064, the first two-point method to calculate the difference in absorbance _{_{obtained: dOD Twopoint =: Abs fit (}} EndPoint) -: Abs fit (StartPoint);

Step S9066-step S90616 are as follows:

Step S9066, within the analysis interval, an N-order polynomial fits the absorbance curve (N≥2), and obtains: Abs _fit ;

Step S9068: Derivate the Abs _fit curve, obtain the tangent equation, and obtain the maximum point MaxPoint;

Step S90610, starting from MaxPoint and extending the linear range with the shortest regression time as a unit;

Step S90612, the integral area between the original absorbance curve and the fastest point tangent is obtained, and the maximum linear range less than the set threshold is obtained;

Step S90614, obtaining the tangent equation Abs _cut and the limit points of the linear range: Cut _Min and Cut _Max ;

Step S90616, the second absorbance difference is calculated by the rate method: dOD _RateMethod = Abs _cut (Cut _Max )-Abs _cut (Cut _Min );

After obtaining a first absorbance and the second absorbance difference _dOD Twopoint difference _dOD RateMethod Through the above steps, perform step S908, the weight calculation i.e. the final absorbance difference is a weighted average performed by assigning a weight function.

Step S908, the obtained according to a certain ratio of the final absorbance difference _{dOD = dOD Twopoint * prop (dOD} ) + dOD RateMethod * (1-prop (dOD)).

The algorithm used in the embodiment of the present application has both the advantages of the two-point method and the rate method. Through the experiment on 4 samples, the two-point method and the rate method are used to calculate the repeatability calculation results of the low value part of the absorbance difference, as shown in Figure 10a, Figure 10b, Figure 10c, Figure 10d in Figure 10; it can be seen that the noise interference caused by the circuit noise When there are fluctuations, the two-point method can obtain good repeatability through the fitting process, and the rate method will be strongly disturbed, resulting in poor repeatability.

If the algorithm of the embodiment of the present application is used, the low value repeatability is basically consistent with the calculation result calculated by the two-point method. Table 1 shows the calculation results of the difference in absorbance obtained by performing experiments on 10 samples. It can be seen that the calculation results of the algorithm using the embodiment of the present application are basically consistent with the two-point method.

Calculate the difference in absorbance by performing experiments on multiple samples of different concentrations. The linear calculation result of the high-value part is shown in Figure 11. Figure 11 shows that when the difference in absorbance is high, the two-point method, the rate method, and the use of this application A comparison diagram of the difference in absorbance calculated by the measurement method of the difference in absorbance of the sample. The concentration of DD in Figure 11 represents the concentration of DD (D-dimer); the difference in absorbance calculated by the two-point method tends to be saturated or even decreased, and the difference in absorbance calculated by the rate method remains monotonously increasing, effectively improving the linear measurement For the range, the linear measurement range of the algorithm used in the embodiment of the present application is basically the same as the rate method.

Table 1 Low-value repeatability calculation results

In some embodiments, the sample analyzer may provide three modes, one mode is calculated using the two-point method, one mode is calculated using the rate method, and one mode is calculated using the measurement method of the sample absorbance difference in the embodiment of the present application , For users to choose.

In some optional embodiments of the present application, the present application also provides a sample analyzer, and the corresponding working method flowchart is shown in FIG. 12. The working method includes the following steps:

Step S1202, the user selects whether to use the algorithm of the embodiment of the present application, if yes, then execute step S1204, if not, execute step S1210;

Step S1204, the instrument obtains configuration parameters;

Step S1206, the instrument carries out experiments, and uses the two-point method and the rate method to calculate the difference in absorbance;

Step S1208, the instrument calculates the difference in absorbance according to the method of the embodiment of the present application;

In step S1210, the user selects another algorithm; other algorithms here may include a two-point method or a rate method, and so on.

Step S1212, the instrument carries out an experiment, and calculates the difference in absorbance according to the method selected by the user;

Step S1214, the user obtains the detection result.

The user needs to choose whether to use the algorithm of the embodiment of the present application. If it is not used, the instrument will carry out the experiment according to the calculation method selected by the user (such as the two-point method or the rate method) and report the results. If used, you need to obtain configuration parameters further. Then, the instrument carries out experiments, and uses the two-point method and the rate method to calculate the difference in absorbance at the same time, and calculates the final difference in absorbance according to the algorithm provided in the embodiment of the present application. Finally, the instrument reports the final difference in absorbance and obtains the test result based on the difference in absorbance, so that the user can obtain the test result, such as the sample concentration. The sample concentration here is an abbreviation defined for brevity, and can be understood as the concentration of a specific substance in the sample, for example, it can be the concentration of D-dimer (DD), fibrin (pro) degradation product (FDP), etc.

In some optional embodiments of the present application, after determining the weights of the first absorbance difference and the second absorbance difference satisfying the weighted average function according to the preset function relationship and the final absorbance difference, the following steps may also be performed: The sample concentration corresponding to the final absorbance difference is determined according to a preset correspondence, wherein the preset correspondence is the correspondence between the absorbance difference and the sample concentration. The sample concentration is in correspondence with the difference in absorbance, and this correspondence is pre-stored in the database. Once the difference in absorbance is determined, the sample concentration can be determined according to this correspondence. Since it belongs to the prior art, it will not be repeated here.

In some optional embodiments of the present application, the above method is applied to a sample analyzer, such as a blood analyzer, a biochemical analyzer, an immunoassay analyzer, a blood coagulation analyzer, and other in vitro diagnostic equipment.

In the embodiment of the present application, by acquiring the absorbance data of the sample object; calculating the first absorbance difference using the two-point method according to the absorbance data, and calculating the second absorbance difference using the rate method according to the absorbance data; combining the first absorbance difference and the second absorbance difference Substitute the preset weighted average function used to calculate the final absorbance difference; determine the weight of the first absorbance difference and the second absorbance difference satisfying the weighted average function and the final absorbance difference according to the preset function relationship, wherein the preset function relationship It is a function of the weight for the first difference in absorbance, the weight for the second difference in absorbance, and the difference in absorbance. The two-point method is adopted by the low value part to obtain better low value repeatability; the rate method is used to obtain the monotonicity of the calculated value of the difference in absorbance and the higher linear measurement range; the low value and the high value Partially well-connected, there is no technical effect of faults or jumps; thus solving the technical problem of measuring the repeatability of the measurement when the absorbance difference is low, and the linear measurement range when the absorbance difference is high. The set threshold of the above sample analyzer can be set by the user. In addition, the decrease speed or increase speed of the absolute value of the weight difference may be set by the user.

An embodiment of the present application further provides one or more non-volatile computer-readable storage media on which a computer program is stored, and the computer program is executed by the processor to perform the steps in the method for measuring the difference in absorbance of a sample of any of the foregoing embodiments .

Corresponding to the above method for measuring the difference in absorbance of the sample, an embodiment of the present application provides the sample analyzer shown in FIG. 1a. As shown in FIG. 1a, the sample analyzer includes: a light source 132, a detector 134, and a processor 136, wherein, The light source 132 is used to emit a light beam for illuminating the sample;

In some optional embodiments of the present application, the above processor 136 is further configured to: determine the sample concentration corresponding to the final absorbance difference according to a preset correspondence, where the preset correspondence is the correspondence between the absorbance difference and the sample concentration relationship.

In some optional embodiments of the present application, the weighted average function is F (x) = a · x + b · (cx) + d; where a is the second absorbance difference and x is the second absorbance difference Weights, b is the first difference in absorbance, 1-x is the weight of the first difference in absorbance, c is a constant greater than or equal to x, d is a constant greater than or equal to 0, and F (x) is the final difference in absorbance.

In some optional embodiments of the present application, the above-mentioned preset functional relationship may be: within a defined domain, if the absorbance difference is less than a set threshold, the weight of the first absorbance difference is greater than the weight of the second absorbance difference; If the difference in absorbance is greater than the set threshold, the weight of the first difference in absorbance is less than the weight of the difference in second absorbance.

Wherein, for the preset function relationship, refer to the above-mentioned related embodiment corresponding to the method for measuring the absorbance of the sample in FIG. 1b.

In some optional embodiments of the present application, if the absorbance difference is less than the set threshold, the absolute value of the weight difference between the first absorbance difference and the second absorbance difference is negatively correlated with the absorbance difference; If the difference in absorbance is greater than the set threshold, the absolute value of the weight difference is positively related to the difference in absorbance.

In some optional embodiments of the present application, if the difference in absorbance is less than the set threshold, the absolute value of the weight difference will decrease as the difference in absorbance increases, the speed of the decrease in the absolute value of the weight difference and the difference in absorbance Positive correlation. If the difference in absorbance is greater than the set threshold, the absolute value of the weight difference will increase as the difference in absorbance increases, and the rate of increase in the absolute value of the weight difference will be inversely related to the difference in absorbance.

In some optional embodiments of the present application, the set threshold is set by the user. The speed of decreasing or increasing the absolute value of the weight difference is set by the user.

The coagulation analyzer in the above embodiment is used for optically measuring and analyzing the number of specific substances related to blood coagulation / fibrinolysis function and the degree of activity thereof, and the specimen is plasma. The coagulation analyzer of the present embodiment optically measures the specimen by the clotting time method, the chromogenic substrate method, and the immunoturbidimetric method. The coagulation time method used in this embodiment is a measurement method in which the coagulation process of a specimen is used as a detection of a change in transmitted light. Measurement items include PT (prothrombin time), APTT (activated partial thrombin time), TT (thrombin time) and FIB (fibrinogen amount). The measurement items of the chromogenic substrate method include AT-III (antithrombin III), and the measurement items of the immunoturbidimetric method include D-Dimer and FDP.

The blood coagulation analyzer includes: at least one reaction container for providing a reaction place for samples and reagents; a sample volume detection device for performing liquid volume detection on the sample to obtain a measured sample volume of the sample; and a blood coagulation detection device for Perform coagulation detection on the sample processed by the reagent to obtain electrical signal information reflecting the coagulation situation; a processor is used to receive and process the electrical signal information output by the coagulation detection device to obtain the measurement parameters of the sample; wherein, the The processor is also used to execute the method for measuring the difference in absorbance of the sample according to any one of the above embodiments.

The sequence numbers of the above embodiments of the present application are for description only, and do not represent the advantages and disadvantages of the embodiments.

In the above embodiments of the present application, the description of each embodiment has its own emphasis. For a part that is not detailed in an embodiment, you can refer to the related descriptions of other embodiments.

In the several embodiments provided in this application, it should be understood that the disclosed technical content may be implemented in other ways. The device embodiments described above are only schematic. For example, the division of units may be a division of logical functions. In actual implementation, there may be other divisions. For example, multiple units or components may be combined or integrated into Another system, or some features can be ignored, or not implemented. In addition, the displayed or discussed mutual coupling or direct coupling or communication connection may be indirect coupling or communication connection through some interfaces, units or modules, and may be in electrical or other forms.

The units described as separate components may or may not be physically separate, and the components displayed as units may or may not be physical units, that is, they may be located in one place, or may be distributed over multiple units. Some or all of the units may be selected according to actual needs to achieve the purpose of the solution of this embodiment.

In addition, each functional unit in each embodiment of the present application may be integrated into one processing unit, or each unit may exist alone physically, or two or more units may be integrated into one unit. The above integrated unit may be implemented in the form of hardware or software functional unit.

If the integrated unit is implemented in the form of a software functional unit and sold or used as an independent product, it can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of the present application essentially or part of the contribution to the existing technology or all or part of the technical solution can be embodied in the form of a software product, the computer software product is stored in a storage medium , Including several instructions to enable a computer device (which may be a personal computer, server, or network device, etc.) to perform all or part of the steps of the methods of the embodiments of the present application. The foregoing storage media include: U disk, read-only memory (ROM, Read-Only Memory), random access memory (RAM, Random Access Memory), mobile hard disk, magnetic disk or optical disk and other media that can store program code .

The above are only the preferred embodiments of the present application. It should be pointed out that for those of ordinary skill in the art, without departing from the principles of the present application, several improvements and retouches can be made. This is the scope of protection of this application.

Claims

A method for measuring the difference in absorbance of a sample, characterized in that it includes:

Obtain the absorbance data of the sample object;

Applying a two-point method to calculate the first absorbance difference according to the absorbance data, and applying a rate method to calculate the second absorbance difference according to the absorbance data;

Substituting the first absorbance difference and the second absorbance difference into a preset weighted average function used to calculate the final absorbance difference;

Determining the weight of the first absorbance difference and the weight of the second absorbance difference and the final absorbance difference satisfying the weighted average function according to a preset function relationship, wherein the preset function relationship is the weight for the first absorbance difference, The weight of the second difference in absorbance is a function of the difference in absorbance.
The method according to claim 1, wherein after determining the weight of the first absorbance difference and the second absorbance difference satisfying the weighted average function according to a preset function relationship and the final absorbance difference, the method further comprises :

The sample concentration corresponding to the final absorbance difference is determined according to a preset correspondence, wherein the preset correspondence is the correspondence between the absorbance difference and the sample concentration.
The method according to claim 1, wherein the preset functional relationship includes:

Within the set definition domain, if the absorbance difference is less than the set threshold, the weight of the first absorbance difference is greater than the weight of the second absorbance difference; if the absorbance difference is greater than the set threshold, the first absorbance The weight of the difference is less than the weight of the second difference in absorbance.
The method according to claim 3, wherein if the difference in absorbance is less than the set threshold, the absolute value of the difference in weight between the weight of the first difference in absorbance and the weight of the second difference in absorbance is The difference in absorbance is negatively correlated; if the difference in absorbance is greater than the set threshold, the absolute value of the weight difference is positively correlated with the difference in absorbance.
The method according to claim 4, characterized in that, if the absorbance difference is less than the set threshold, the absolute value of the weighted phase difference decreases as the absorbance difference increases, and the absolute value of the weighted phase difference The speed of decrease is positively correlated with the difference in absorbance.
The method according to claim 4, wherein if the difference in absorbance is greater than the set threshold, the absolute value of the weighted phase difference increases as the difference in absorbance increases, and the absolute value of the weighted phase difference The rate of increase is inversely related to the difference in absorbance.
The method according to any one of claims 3 to 6, wherein the set threshold is set by a user.
The method according to claim 5 or 6, wherein the rate of change of the absolute value of the weight difference is set by the user.
The method according to claim 3, wherein the preset functional relationship further comprises:

In the defined domain of the setting, if the difference in absorbance is equal to the set threshold, the weight of the first difference in absorbance and the weight of the second difference in absorbance are the same.
The method of claim 1, wherein the weighted average function is:

F (x) = a · x + b · (c-x) + d;

Where a is the second absorbance difference, x is the weight of the second absorbance difference, b is the first absorbance difference, (cx) is the weight of the first absorbance difference, and c is greater than or equal to x , D is a constant greater than or equal to 0, and F (x) is the final absorbance difference.
The method of claim 1, wherein the method is applied to a sample analyzer.
A sample analyzer, characterized by comprising: a light source, a detector and a processor; wherein,

The light source is used to emit a light beam for illuminating the sample;

The detector is used to detect luminous flux data generated after the light beam illuminates the sample;

The processor runs a program, wherein, when the program runs, the following processing steps are performed on the data output from the detector:

Calculating absorbance data according to the luminous flux data;

Applying a two-point method to calculate the first absorbance difference according to the absorbance data, and applying a rate method to calculate the second absorbance difference according to the absorbance data;

Substituting the first absorbance difference and the second absorbance difference into a preset weighted average function used to calculate the final absorbance difference;

Determining the weight of the first absorbance difference and the weight of the second absorbance difference and the final absorbance difference satisfying the weighted average function according to a preset function relationship, wherein the preset function relationship is the weight for the first absorbance difference, The weight of the second difference in absorbance is a function of the difference in absorbance.
The sample analyzer according to claim 12, wherein the processor is further configured to: determine the sample concentration corresponding to the final absorbance difference according to a preset correspondence, wherein the preset correspondence is absorbance Correspondence between the difference and the sample concentration.
The sample analyzer according to claim 12, wherein the preset functional relationship includes:

Within the set definition domain, if the absorbance difference is less than the set threshold, the weight of the first absorbance difference is greater than the weight of the second absorbance difference; if the absorbance difference is greater than the set threshold, the first absorbance The weight of the difference is less than the weight of the second difference in absorbance.
The sample analyzer according to claim 14, wherein if the difference in absorbance is less than the set threshold, the weight difference between the weight of the first absorbance difference and the weight difference between the second absorbance difference is an absolute value weight difference The absolute value of is negatively correlated with the difference in absorbance; if the difference in absorbance is greater than the set threshold, the absolute value of the weight difference is positively correlated with the difference in absorbance.
The sample analyzer according to claim 15, characterized in that, if the difference in absorbance is less than the set threshold, the absolute value of the weighted phase difference decreases as the difference in absorbance increases, the weighted difference The rate of decrease in absolute value is positively correlated with the difference in absorbance.
The sample analyzer according to claim 15, wherein if the difference in absorbance is greater than the set threshold, the absolute value of the weighted phase difference increases with the increase in the difference in absorbance. The rate of increase in absolute value is inversely related to the difference in absorbance.
The sample analyzer according to any one of claims 14 to 17, wherein the set threshold is set by a user.
The sample analyzer according to claim 16 or 17, wherein the rate of change of the absolute value of the weight difference is set by the user.
The sample analyzer according to claim 12, wherein the weighted average function is:

F (x) = a · x + b · (c-x) + d;

Where a is the second absorbance difference, x is the weight of the second absorbance difference, b is the first absorbance difference, cx is the weight of the first absorbance difference, c is the constant greater than or equal to x, and d is greater than or equal to 0 Constant, F (x) is the final absorbance difference.
One or more non-volatile computer-readable storage media on which a computer program is stored, characterized in that, when the computer program is executed by a processor, the method according to any one of claims 1-11 is implemented step.