CN111579776B - Method, device and system for determining substance concentration through test strip and storage medium - Google Patents
Method, device and system for determining substance concentration through test strip and storage medium Download PDFInfo
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- CN111579776B CN111579776B CN202010486949.2A CN202010486949A CN111579776B CN 111579776 B CN111579776 B CN 111579776B CN 202010486949 A CN202010486949 A CN 202010486949A CN 111579776 B CN111579776 B CN 111579776B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
Abstract
The application provides a method, a device and a system for determining substance concentration through a test strip and a storage medium, and belongs to the technical field of fluorescence immunochromatography. The method comprises the following steps: acquiring a corresponding fluorescence curve in a scanning range on a target test strip through a photoelectric sensor; according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove, the position of a preset line is determined, and the preset line comprises: a quality control line and a detection line; determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line; and determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve. The method and the device can more accurately acquire the relative peak value of the preset line of the fluorescence curve, and further enable the accuracy of the substance concentration of the obtained sample to be detected to be higher.
Description
Technical Field
The application relates to the technical field of fluorescence immunochromatography, in particular to a method, a device and a system for determining substance concentration through a test strip and a storage medium.
Background
The fluorescence immunochromatography technology detects whether the antigen or the antibody to be detected exists in the sample solution through test strip chromatography and gives qualitative or quantitative indexes of the substance to be detected.
At present, the main test method is to put the test strip into a card holder and obtain the fluorescence value on the test strip by an optical detection device to form a fluorescence curve.
However, in the current testing process, the position of the actual scanning card holder is usually deviated due to the deviation of the position of the test strip card holder, which also causes the deviation of the peak position of the formed fluorescence curve, and once the deviation is too large from the target position, the peak searching error is caused, thereby causing the wrong qualitative or quantitative index of the obtained substance to be tested.
Disclosure of Invention
The application aims to provide a method, a device, a system and a storage medium for determining the concentration of a substance through a test strip, so that the peak position of a preset line of a fluorescence curve can be more accurately obtained, and the accuracy of qualitative or quantitative indexes of an antigen or an antibody to be detected in an obtained sample is higher.
The embodiment of the application is realized as follows:
in one aspect of the embodiments of the present application, a method for determining a concentration of a substance by a test strip is provided, where the method includes:
acquiring a corresponding fluorescence curve in a scanning range on a target test strip through a photoelectric sensor;
according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove, the position of a preset line is determined, and the preset line comprises: a quality control line and a detection line;
determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line;
and determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve.
Optionally, determining the position of the preset line according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove, including:
preprocessing the fluorescence curve to obtain a first derivative curve of the fluorescence curve;
determining the position of a rising edge in the fluorescence curve according to the first derivative curve;
according to the position of rising edge and the rising edge interval distance of test paper strip default groove, confirm the position of predetermineeing the line, the rising edge interval distance of test paper strip default groove includes: the preset distance from the rising edge of the test strip groove to the quality control line and the preset distance from the rising edge of the test strip groove to the detection line.
Optionally, determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line includes:
determining the fluorescence peak value of the preset line and the fluorescence value of the comparison base line of the preset line according to the position of the preset line;
and determining the relative peak value of the preset line according to the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line.
Optionally, determining the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line according to the position of the preset line includes:
acquiring a fluorescence peak value of a preset line corresponding to the position of the preset line;
respectively determining baselines on two sides of the preset line according to the fluorescence peak value of the preset line and the distance between the preset baselines;
and selecting the corresponding base line with a higher fluorescence value from the base lines on the two sides of the preset line as a comparison base line, and recording the fluorescence value of the comparison base line.
Optionally, determining a corresponding peak value of the preset line according to the peak value of the preset line and the fluorescence value of the comparison baseline of the preset line, includes:
and taking the difference value of the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline as the relative peak value of the preset line.
Optionally, determining the substance concentration of the sample to be tested according to the relative peak value of the fluorescence curve, comprising:
judging whether the detection result of the test strip is effective or not according to the fluorescence peak value of the quality control line;
and if the detection result is valid, determining the substance concentration of the sample to be detected according to the relative peak value of the detection line.
In another aspect of the embodiments of the present application, there is provided a device for determining a concentration of a substance by a test strip, the device including: the device comprises an acquisition module, a position determination module, a peak value determination module and a concentration determination module;
the acquisition module is used for acquiring and acquiring a corresponding fluorescence curve in a scanning range on the target test strip through the photoelectric sensor;
the position determination module is used for determining the position of a preset line according to the position of a rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove, and the preset line comprises: a quality control line and a detection line;
the peak value determining module is used for determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line;
and the concentration determination module is used for determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve.
Optionally, the position determining module is specifically configured to preprocess the fluorescence curve to obtain a first derivative curve of the fluorescence curve; determining the position of a rising edge in the fluorescence curve according to the first derivative curve; according to the position on rising edge and the rising edge interval distance of test paper strip default groove, confirm the position of predetermineeing the line, the rising edge interval distance of test paper strip default groove includes: the preset distance from the rising edge of the test strip groove to the quality control line and the preset distance from the rising edge of the test strip groove to the detection line.
Optionally, the peak determining module is specifically configured to determine a fluorescence peak value of the preset line and a fluorescence value of the comparison baseline of the preset line according to the position of the preset line; and determining the relative peak value of the preset line according to the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line.
Optionally, the peak determining module is specifically configured to obtain a fluorescence peak of the preset line corresponding to the position of the preset line; respectively determining baselines on two sides of the preset line according to the fluorescence peak value of the preset line and the distance between the preset baselines; and selecting the corresponding base line with a higher fluorescence value from the base lines on the two sides of the preset line as a comparison base line, and recording the fluorescence value of the comparison base line.
Optionally, the peak determining module is specifically configured to use a difference between a fluorescence peak value of the preset line and a fluorescence value of the comparison baseline as a relative peak value of the preset line.
Optionally, the concentration determination module is specifically configured to determine whether a detection result of the test strip is valid according to a fluorescence peak value of the quality control line; and if the detection result is valid, determining the substance concentration of the sample to be detected according to the relative peak value of the detection line.
In another aspect of the embodiments of the present application, there is provided a system for determining a concentration of a substance by a test strip, the system including: optical detection equipment, circuit equipment and test paper strips; the circuit device detects the test strip by the optical detection device using the steps of the method for determining the concentration of the substance by the test strip as described above.
In another aspect of the embodiments of the present application, there is provided a circuit device, including: the device comprises a memory and a processor, wherein a computer program capable of running on the processor is stored in the memory, and when the processor executes the computer program, the steps of the method for determining the concentration of the substance through the test strip are realized.
In another aspect of the embodiments of the present application, a storage medium is provided, and the storage medium stores a computer program, and the computer program, when executed by a processor, implements the steps of the method for determining a concentration of a substance by a test strip.
The beneficial effects of the embodiment of the application include:
according to the method, the device, the system and the storage medium for determining the substance concentration through the test strip, the corresponding fluorescence curve in the scanning range of the target test strip is acquired through the photoelectric sensor, the position of the preset line can be more accurately determined according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the preset grooves of the test strip, the situation that the position of the preset line is inaccurate due to the dislocation of the clamping supports of the test strip is prevented, the relative peak value of the preset line of the fluorescence curve is determined according to the position of the preset line, the relative peak value of the preset line of the obtained fluorescence curve can be more accurate, the substance concentration of the sample to be detected is determined according to the relative peak value of the preset line of the fluorescence curve, and the accuracy of the substance concentration of the sample to be detected can be higher.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained from the drawings without inventive effort.
FIG. 1 is a schematic diagram of a system for determining a concentration of a substance via a test strip according to an embodiment of the present disclosure;
fig. 2 is a schematic structural diagram of a test strip provided in an embodiment of the present application;
FIG. 3 is a diagram of a test strip in the form of a single card;
fig. 4 is a schematic view of a scanning position of the test strip provided in the embodiment of the present application;
FIG. 5 is a schematic flow chart illustrating a method for determining a concentration of a substance by a test strip according to an embodiment of the present disclosure;
FIG. 6 is a schematic representation of a fluorescence curve provided in an embodiment of the present application;
FIG. 7 is a schematic flowchart of determining a position of a default line according to an embodiment of the present disclosure;
FIG. 8 is a schematic diagram of the first derivative of a fluorescence curve provided in an embodiment of the present application;
FIG. 9 is a schematic diagram of a process for determining the relative peak value of a predetermined line of a fluorescence curve according to an embodiment of the present application;
FIG. 10 is a schematic flow chart illustrating the process of determining the fluorescence peak value of the predetermined line and the fluorescence value of the comparative baseline of the predetermined line according to the embodiment of the present application;
FIG. 11 is a schematic diagram of a process for determining a substance concentration of a sample to be tested according to an embodiment of the present disclosure;
FIG. 12 is a schematic view of an apparatus for determining a concentration of a substance via a test strip provided in an embodiment of the present application;
fig. 13 is a schematic structural diagram of a circuit device according to an embodiment of the present application.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are some embodiments of the present application, but not all embodiments. The components of the embodiments of the present application, generally described and illustrated in the figures herein, can be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present application, as presented in the figures, is not intended to limit the scope of the claimed application, but is merely representative of selected embodiments of the application. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments in the present application without making any creative effort belong to the protection scope of the present application.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
Furthermore, the terms "first," "second," "third," and the like are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance.
The following is an explanation of a system applied to the method for determining the concentration of a substance by a test strip, which is adopted in the present application, by a plurality of examples.
Fig. 1 is a schematic diagram of a system for determining a substance concentration by a test strip according to an embodiment of the present application, please refer to fig. 1, where the system includes: a circuit device 10, an optical detection device 20 and a test strip 30; the optical detection device 20 detects the fluorescence value on the test strip 30, and outputs a fluorescence curve after AD sampling and amplification by the circuit device 10.
Wherein the optical detection device 20 comprises: a first convex lens 21 and a second convex lens 22 which are arranged opposite to each other, a first optical filter 23, a second optical filter 24, a dichroic mirror 25, a light source 26, and a photoelectric sensor 27. Wherein the first filter 23 is disposed between the first convex lens 21 and the dichroic mirror 25, and the second filter 24 is disposed between the light source 26 and the dichroic mirror 25; light generated by the light source 26 can be incident on the dichroic mirror 25 through the second optical filter 24, the photoelectric sensor 27 is further disposed on the flat transparent side of the first convex lens 21, and the test strip 30 is further disposed on the flat transparent side of the second convex lens 22, wherein the test strip 30 can be a test strip disposed in a test strip holder.
It should be noted that light emitted from the light source 26 is filtered by the second optical filter 24, and then enters the dichroic mirror 25, and is reflected by the dichroic mirror 25, the dichroic mirror 25 is a long-pass dichroic mirror, long waves pass through, and short waves are reflected, and the reflected light is focused to the test strip 30 by the second convex lens 22, fluorescent microspheres on the test strip 30 are excited to generate excitation light, the wavelength of the excitation light is greater than that of the emitted light, the excitation light passes through the dichroic mirror 25, and is focused to the photoelectric sensor 27 by the first convex lens 21 after being filtered by the first optical filter 23, the photoelectric sensor 27 converts an optical signal into an electrical signal, the circuit device 10 performs AD sampling (analog-to-digital sampling) on the electrical signal, so as to convert analog quantity information of the electrical signal into corresponding digital quantity information, and further generate a corresponding fluorescence curve according to the digital quantity information, where the fluorescence curve can reflect the size of a fluorescence value at each position on the test strip.
Alternatively, the first filter 23 may be used to select optical signals conforming to the radiation band of long-wavelength optical signals, and the second filter 24 may be used to select optical signals conforming to the radiation band of short-wavelength optical signals; the first convex lens 21 and the second convex lens 22 are used for converging light rays.
The following explains the specific structure of the test strip 30 and the working principle of the test strip 30 by specific embodiments.
Fig. 2 is a schematic structural diagram of a test strip provided in an embodiment of the present application, please refer to fig. 2, where the test strip 30 includes: the kit comprises a base plate 31, a nitrocellulose membrane (NC membrane 32), a marker strip 33, a sample pad 34 and absorbent paper 35, wherein the nitrocellulose membrane 32 is coated with: quality control line (C line 36), detection line (T line 37).
The test strip 30 can be applied to the fluorescence immunochromatography technique, which generally includes a sandwich method and a competition method, and the sandwich method includes a double-antigen sandwich method and a double-antibody sandwich method, in this embodiment, the double-antibody sandwich method is taken as an example, first, a sample liquid to be tested is centrifugally diluted and then is added to the sample pad 34, and enters the label strip 33 through chromatography, an antigen in the sample liquid to be tested and the antibody 1 marked by the fluorescent microsphere on the label strip 33 are specifically combined to form an "antigen-antibody 1-fluorescent microsphere" structure, and then, chromatography is continued on the NC membrane 32. NC membrane 32 is pre-coated with 2 antibody lines of 1mm width, which are: the antibody 2 on the left side can be specifically combined with the detected antigen in the sample liquid to be detected to form a double-antibody sandwich structure of an antibody 2-antigen-antibody 1-fluorescent microsphere, namely the T line 37; the antibody 3 on the right side can be combined with the antibody 1 on the label strip 33 to form a secondary antibody structure of "antibody 3-antibody 1-fluorescent microsphere", namely, the C-line 36. When the liquid chromatography of the sample to be tested reacted with the label strip 33 passes through the 2 antibody lines, the sample will be combined with the 2 antibody lines and captured. Since the antibody 1 on the label strip 33 is labeled with the fluorescent microsphere, the antibody line will generate fluorescence under excitation of excitation light with a specific wavelength. Wherein, when the tested antigen exists in the sample liquid to be tested, the T line 37 generates a fluorescence signal, and the more the antigen, the stronger the fluorescence signal. Regardless of the presence or absence of antigen in the sample fluid, a fluorescent signal appears at the location of the C-line 36 after chromatography of the sample fluid through the C-line 36. The C line 36 shows that the liquid chromatography of the sample to be detected is normal and the antibody 1 and the fluorescent microsphere are marked normally, so that the quality monitoring of the immunochromatography detection is realized. After the sample solution passes through the above two antibody lines, the unreacted components and the unbound "antibody 1-fluorescent microsphere" structure will continue to be chromatographed until all is absorbed by the absorbent paper 35. Therefore, during the whole immunochromatography test, all the added sample liquid to be tested should sequentially pass through the sample pad 34, the label strip 33, the T line 37 and the C line 36, and be finally absorbed by the absorbent paper 35. Usually, the process is completed within 5 to 30 minutes according to the difference of the fluorescence immunochromatographic test strip. The bottom plate 31 is fixed below the NC film 32, the marker strip 33, the sample pad 34, and the absorbent paper 35, and fixes the entire test strip 30.
Fig. 3 is a diagram of a test strip single card provided in an embodiment of the present application, please refer to fig. 2 and fig. 3 in combination, where a test strip 30 can be loaded into the test strip single card 40, and the test strip single card 40 includes: test paper groove 41 and sample groove 44, test paper groove 41's both ends include respectively: a rising edge 42 of the trough, and a falling edge 43 of the trough.
It should be noted that, the test strip groove 41 may be correspondingly loaded with the NC film 32 portion of the test strip 30, the rising edge 42 of the groove may correspond to the left end of the NC film 32 in fig. 2, the falling edge 43 of the groove may correspond to the right end of the NC film 32 in fig. 2, and the sample groove 44 may correspond to the sample pad 34 in the test strip 30 for adding the sample solution to be tested.
Fig. 4 is a schematic diagram of scanning positions of test strips provided in an embodiment of the present application, please refer to fig. 2 and fig. 4 in combination, a plurality of test strips 30 can be respectively loaded into a test strip quadruple card 53, the test strip quadruple card 53 is embedded into a card holder 52, the card holder 52 is supported by a drawer 51, the drawer 51 can move left and right to execute an entry and exit, and the entry is performed in a scanning range in which the test strip quadruple card 53 and the card holder 52 are loaded for optical detection, where a scanning start line 54 is located on the left side of a test strip slot of the test strip quadruple card 53, and a scanning end line 55 is located on the right side of the test strip slot of the test strip quadruple card 53.
Referring to fig. 1 and 4, before the scanning is started, the test strip quadruple card 53 and the card holder 52 are held on the drawer 51 for entering the cabin. When scanning starts, the motor drives the optical detection device 20 to move from the scanning start line 54 to the scanning end line 55 at a fixed speed, so as to complete the process of detection scanning, the photoelectric sensor 27 can convert the received optical signal and send the converted electrical signal to the circuit device 10, and the circuit device 10 performs AD sampling amplification according to the electrical signal to output a corresponding fluorescence curve, wherein the ordinate of the fluorescence curve identifies the magnitude of the fluorescence value, and the abscissa represents the position in the test strip, which can be represented by numerical values, for example: the position of the scanning start line 54 is recorded as 0, and the distance is obtained according to a certain distance unit (for example: millimeter), so as to record the position of the whole detection scanning process.
The specific process of the method for determining the concentration of a substance by means of a test strip is explained below by means of specific examples.
Fig. 5 is a schematic flowchart of a method for determining a substance concentration by a test strip according to an embodiment of the present application, please refer to fig. 5, where the method includes:
s10: and acquiring a fluorescence curve in a scanning range on the target test strip through the collection of the photoelectric sensor.
It should be noted that, emitted light generated by the test strip excited by the excitation light can be collected by the photoelectric sensor, and the optical signal can be converted into a corresponding electrical signal, and AD sampling can be performed according to the electrical signal, that is, analog quantity information of the electrical signal can be converted into corresponding digital quantity information, and a corresponding fluorescence curve can be drawn according to the digital quantity information, and the fluorescence curve can reflect the size of the fluorescence value at each position in the scanning range on the test strip. The target test strip can be a single card in fig. 3 or a 4-card in fig. 4, and a user can select different items to perform detection scanning according to actual requirements, so that a fluorescence curve can be obtained.
S20: and determining the position of the preset line according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove.
Wherein, predetermine the line and include: a quality control line (C line) and a detection line (T line).
It should be noted that the position of the preset line is the corresponding position of the quality control line and the detection line in the fluorescence curve of the test strip; the position of the rising edge of the test strip groove is the corresponding position of the rising edge of the test strip clamping groove in the fluorescence curve; the distance between the rising edges of the preset grooves can be the distance from the rising edges of the grooves to a preset line, and the distances between the rising edges of the preset grooves are equal for the same type of test strips. Alternatively, the distance between the rising edges of the preset slots may be obtained in a certain distance unit (e.g., mm) according to the above-mentioned position corresponding to the scanning start line as 0.
In the fluorescence curve, the abscissa represents the position of the test strip, and after the position of the rising edge of the slot in the fluorescence curve is determined, the result obtained by adding the position of the rising edge of the slot and the distance between the rising edges of the preset slots is the position of the preset line, and the specific calculation mode is as follows:
P 0 +L＝P；
wherein, P 0 L is the distance of the rising edge of the preset groove, and P is the position of the preset line.
S30: and determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line.
It should be noted that, after the position of the preset line is determined in the fluorescence curve, the ordinate corresponding to the position is found, and within a certain range of the preset line, the maximum fluorescence value is found as the fluorescence peak value of the preset line, and then the relative peak value of the preset line of the fluorescence curve is obtained according to the relative height of the fluorescence peak value in the fluorescence curve, where the relative height may be the distance between the peak top end corresponding to the fluorescence peak value and both ends of the peak in the fluorescence curve.
Since the preset line may include the C line and the T line, the obtained relative peak value may also include the relative peak value of the C line and the relative peak value of the T line.
S40: and determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve.
It should be noted that, after determining the relative peak value of the preset line of the fluorescence curve, if the relative peak value is a quantitative item, the T-line relative peak value may be converted into the concentration of the substance to be detected in the sample to be detected by a preset formula, where the preset formula may be the corresponding relationship between the T-line relative peak value and the substance concentration obtained by a large number of experiments by a worker; if the relative peak value is a qualitative item, the relative peak value can be converted into a qualitative index of the substance to be detected in the sample through a preset reference range, and the preset reference range is a corresponding relation between the relative peak value and the qualitative index obtained through a large number of clinical statistics.
Alternatively, in the process of calculating the quantitative concentration or qualitative index of the substance to be detected in the sample, the relative peak value corresponding to the T-line fluorescence peak value can be used to determine the substance concentration of the sample to be detected.
According to the method for determining the substance concentration through the test strip, the fluorescence curve in the scanning range of the target test strip is acquired through the photoelectric sensor, the position of the preset line can be more accurately determined according to the position of the rising edge of the groove in the fluorescence curve and the distance between the rising edges of the preset grooves of the test strip, the situation that the position of the preset line is inaccurate due to dislocation of the test strip card in the drawer is prevented, the correctness of the relative peak value of the preset line of the fluorescence curve is further ensured by ensuring the accuracy of the position of the preset line, the substance concentration of the sample to be determined is determined according to the relative peak value of the T line of the fluorescence curve, and the accuracy of the substance concentration of the obtained sample to be determined can be ensured.
The above fluorescence curve is explained below by specific examples.
Fig. 6 is a schematic view of a fluorescence curve provided in an embodiment of the present application, please refer to fig. 2, fig. 3, fig. 4, and fig. 6 in combination, where fig. 6 includes a T-line fluorescence peak, a C-line fluorescence peak, a chromatographic upper edge being a position corresponding to a rising edge of a trough, and a chromatographic lower edge being a position corresponding to a falling edge of the trough.
A certain distance is left between the scanning start line 54 and the rising edge 42 of the groove, the positions corresponding to the test paper grooves 41 are all the NC membrane 32, the fluorescent microspheres flow on the NC membrane 32 along with the sample liquid, and even after complete flow, a small amount of free fluorescent microspheres still remain on the NC membrane 32. At the beginning, the fluorescence value of the test strip 30 is small, and when the fluorescent microspheres on the NC film 32 are scanned, the fluorescence value has a gradually increasing rising edge. Similarly, if the entire test paper slot 41 is subjected to chromatography and the entire NC membrane 32 has a distribution of fluorescent microspheres, there is a decreasing falling edge of the fluorescence value at the position from the NC membrane 32 to the falling edge 43 of the slot. For example, in the double antibody sandwich method, if the sample liquid to be detected contains the antigen to be detected, the antibody 2 at the position of the T line 37 is combined with the antigen-antibody conjugate, and the antigen-antibody conjugate is combined on the fluorescent microsphere, the more the antigen is, the higher the concentration of the fluorescent microsphere at the position of the T line 37 is, and the higher the fluorescence value is. The antibody 3 at the C line 36 is bound with the free antibody 1 after removing the bound antigen to be detected, so that the more the antigen is, the less the free antibody 1 bound with the fluorescent microsphere is, the lower the fluorescence value is, but the minimum limit level must be higher to ensure the sufficiency of the antigen reaction in the sample liquid.
The specific steps for determining the position of the preset line are explained below by means of specific embodiments.
Fig. 7 is a schematic flowchart of a process of determining a position of a preset line according to an embodiment of the present application, please refer to fig. 6 and 7 in combination, and S20: according to the position of the rising edge of the groove in the fluorescence curve and the distance between the rising edges of the preset grooves, the position of a preset line is determined, and the method comprises the following steps:
s210: and preprocessing the fluorescence curve to obtain a first derivative curve of the fluorescence curve.
It should be noted that after the fluorescence curve is obtained, the fluorescence curve may be preprocessed, and the preprocessing may be performed by performing a first derivation on the fluorescence curve, so as to obtain a first derivative curve corresponding to the fluorescence curve.
Specifically, the rising edge in the fluorescence curve is positive in its first derivative curve, and the falling edge in the fluorescence curve is negative in its first derivative curve.
Fig. 8 is a schematic diagram of a first derivative of a fluorescence curve provided in an embodiment of the present application, please refer to fig. 8, where the first derivative curve in fig. 8 is obtained by first derivative of the fluorescence curve in fig. 6. Referring to fig. 7 and 8 in combination, how to determine the position of the predetermined line according to the first derivative curve in fig. 8 is explained below by a specific embodiment.
S220: the position of the rising edge of the trough in the fluorescence curve is determined from the first derivative curve.
According to the position of the initial N points (N is the number of the preset points, N is a positive integer greater than 0) in the fluorescence curve in fig. 6 as the front baseline, when the fluorescence value of the curve is continuously greater than the front baseline for the first time, it may be determined in the first derivative curve in fig. 8 whether the first derivative of the fluorescence value curve in this section is a positive number, if the first derivative is a positive number, the first derivative is the chromatographic upper edge, and the position of the chromatographic upper edge, that is, the position of the chromatographic upper edge, is the position of the chromatographic upper edge of the channel.
S230: and determining the position of the preset line according to the position of the rising edge of the groove and the distance between the rising edges of the preset grooves.
Wherein, the rising edge interval distance of presetting the groove includes: the preset distance from the rising edge of the groove to the quality control line and the preset distance from the rising edge of the groove to the detection line.
The specific calculation method for the position of the line C is as follows:
P 0 +L C ＝P C ；
wherein L is C Is a predetermined distance, P, from the rising edge of the trough to the quality control line C Is the position of line C.
The specific calculation method for the position of the T line is as follows:
P 0 +L T ＝P T ；
wherein L is T For a predetermined distance, P, from the rising edge of the trough to the detection line T The position of the T-line.
The procedure for determining the relative peak of the predetermined line of the fluorescence curve is explained below by means of a specific example.
Fig. 9 is a schematic flow chart illustrating the process of determining the relative peak value of the preset line of the fluorescence curve according to the embodiment of the present application, please refer to fig. 9, S30: determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line, comprising the following steps:
s310: and determining the fluorescence peak value of the preset line and the fluorescence value of the comparison base line of the preset line according to the position of the preset line.
It should be noted that, after the position of the preset line is determined, the fluorescence peak value of the preset line may be determined within a certain preset range of the preset line, where the preset range may be within a certain preset distance around the preset line, and the fluorescence peak value may be a value with the highest fluorescence value in the preset range.
Optionally, the comparison baseline may be a position of a peak corresponding to the fluorescence peak value at one of two ends of the peak in the fluorescence curve, and the fluorescence value at the position is the fluorescence value of the comparison baseline of the preset line. Each preset line may have a different comparison baseline, such as: the comparison baseline of the T-line may be a position of a peak corresponding to the fluorescence peak value of the T-line at one of two ends of the peak in the fluorescence curve, and the determination method of the comparison baseline position of the C-line is similar to the T-line, which is not described herein again.
S320: and determining the relative peak value of the preset line according to the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line.
It should be noted that the relative peak value may be calculated according to the fluorescence value of the comparison baseline between the fluorescence peak value and the preset line, and the relative peak height of the peak corresponding to the fluorescence peak value may also have a fluorescence value in a certain range around the preset line, so the size of the fluorescence peak value may not represent the peak height of the peak, and it is usually necessary to obtain the relative peak height of the peak to perform corresponding calculation.
The specific procedures for obtaining a comparison baseline and determining relative peak values are explained below by specific examples.
Fig. 10 is a schematic flow chart illustrating the process of determining the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line according to the embodiment of the present application, please refer to fig. 10, S310: determining the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line according to the position of the preset line, wherein the method comprises the following steps:
s311: and acquiring the fluorescence peak value of the preset line corresponding to the position of the preset line.
It should be noted that, in this embodiment, the distance from the rising edge of the slot to the T line is 4 mm, the distance to the C line is 10 mm, the speed of the motor driving the optical detection device to scan the test strip is fixed, 56 fluorescence values are sampled at 1mm, and the tolerance of the test strip position is 1 mm. Starting from the rising edge position of the test strip groove, the maximum fluorescence value in the range of 3-5 mm is the fluorescence peak value corresponding to the T line, and the maximum fluorescence value in the range of 9-11 mm is the fluorescence peak value corresponding to the C line.
S312: and respectively determining baselines on two sides of the preset line according to the fluorescence peak value of the preset line and the distance between the preset baselines.
In this embodiment, the T-line and the C-line themselves are 1mm wide, and the minimum of the fluorescence values within 2 mm to the left of the fluorescence peak can be used as the left baseline, and the minimum of the fluorescence values within 2 mm to the right of the fluorescence peak can be used as the right baseline.
S313: and selecting the baseline with higher fluorescence value corresponding to the baselines on the two sides of the preset line as a comparison baseline, and recording the fluorescence value of the comparison baseline.
Taking the position relation in the figure as reference, the baselines at two sides of the preset line can be recorded as a left base line and a right base line, the fluorescence values corresponding to the left base line and the right base line can be compared, the position corresponding to the larger fluorescence value is selected as the comparison base line, and the fluorescence value of the comparison base line is recorded. For example: if the fluorescence value of the left baseline is 2400 and the fluorescence value of the right baseline is 2600, the right baseline can be selected as the comparison baseline.
Optionally, S320: determining the corresponding peak value of the preset line according to the peak value of the preset line and the fluorescence value of the comparison base line of the preset line, wherein the method comprises the following steps:
and taking the difference value of the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline as the relative peak value of the preset line.
The specific calculation formula is as follows:
ΔF＝F M -F B ；
wherein, delta F is the relative peak value of a preset line, F M To preset the fluorescence peak of the line, F B Fluorescence values for the comparison baseline for the preset line.
The specific procedure for determining the concentration of a substance in a sample to be tested is explained below by means of a specific example.
Fig. 11 is a schematic flowchart of a process for determining a substance concentration of a sample to be tested according to an embodiment of the present application, please refer to fig. 11, S40: determining the substance concentration of the sample to be detected according to the relative peak value of the fluorescence curve, comprising the following steps:
s410: and judging whether the detection result of the test strip is effective or not according to the fluorescence peak value of the quality control line.
It should be noted that, regardless of whether an antigen exists in the sample liquid to be detected, a fluorescence signal appears at the position of the C line after the sample liquid chromatography passes through the C line. The C line shows that the liquid chromatography of the sample to be detected is normal and the antibody 1 and the fluorescent microsphere are marked normally, so that the quality monitoring of the immunochromatography detection is realized. Therefore, whether the test result of the test strip is valid can be judged by judging whether the relative peak value of the C line is normal or not, namely whether the fluorescence value meets a certain preset condition or not. If the fluorescence value of the C line position does not reach the preset condition, the detection result is invalid, and a new test strip needs to be used for detection scanning so as to ensure the accuracy of the finally obtained substance concentration.
S420: and if the detection result is valid, determining the substance concentration of the sample to be detected according to the relative peak value of the detection line.
It should be noted that, when the antigen to be detected exists in the sample liquid to be detected, a fluorescence signal appears on the T-line, and the more the antigen is, the stronger the fluorescence signal is, taking the quantitative item as an example, the substance concentration obtained by calculating the relative peak value of the T-line through the following calculation formula is the substance concentration of the sample to be detected, and the specific formula is:
wherein y is a fluorescence value, which is a relative peak value of the T-line in this embodiment, x is a substance concentration of the sample to be measured, and a, B, C, and D are preset constant parameters, and the substance concentration of the sample to be measured can be determined according to the relative peak value of the fluorescence curve by the formula.
The following explains a specific structure of the device for determining the concentration of a substance by a test strip provided in this embodiment by a specific embodiment.
Fig. 12 is a schematic view of an apparatus for determining a substance concentration by a test strip according to an embodiment of the present application, please refer to fig. 12, the apparatus includes: the device comprises an acquisition module 100, a position determination module 200, a peak value determination module 300 and a concentration determination module 400;
the acquisition module 100 is used for acquiring and acquiring a corresponding fluorescence curve in a scanning range on a target test strip through a photoelectric sensor;
the position determination module 200 is configured to determine a position of a preset line according to a position of a rising edge of a groove in the fluorescence curve and a distance between rising edges of the test strip preset groove, where the preset line includes: a quality control line and a detection line;
a peak value determining module 300, configured to determine a relative peak value of a preset line of the fluorescence curve according to a position of the preset line;
and the concentration determining module 400 is used for determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve.
Optionally, the position determining module 200 is specifically configured to preprocess the fluorescence curve to obtain a first derivative curve of the fluorescence curve; determining the position of the rising edge of the groove in the fluorescence curve according to the first derivative curve; according to the rising edge position in groove and the rising edge interval distance in the test paper strip preset groove, the position of the preset line is determined, and the rising edge interval distance in the preset groove comprises: the preset distance from the rising edge of the groove to the quality control line and the preset distance from the rising edge of the groove to the detection line.
Optionally, the peak determining module 300 is specifically configured to determine a fluorescence peak value of the preset line and a fluorescence value of the comparison baseline of the preset line according to the position of the preset line; and determining the relative peak value of the preset line according to the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line.
Optionally, the peak determining module 300 is specifically configured to obtain a fluorescence peak of the preset line corresponding to the position of the preset line; respectively determining baselines on two sides of the preset line according to the fluorescence peak value of the preset line and the distance between the preset baselines; and selecting the corresponding base line with a higher fluorescence value from the base lines on the two sides of the preset line as a comparison base line, and recording the fluorescence value of the comparison base line.
Optionally, the peak determining module 300 is specifically configured to use a difference between a fluorescence peak value of the preset line and a fluorescence value of the comparison baseline as a relative peak value of the preset line.
Optionally, the concentration determining module 400 is specifically configured to determine whether the detection result of the test strip is valid according to a fluorescence peak value of the quality control line; and if the detection result is valid, determining the substance concentration of the sample to be detected according to the relative peak value of the detection line.
The embodiment of the application provides a device for determining substance concentration through test paper strip, acquire the fluorescence curve that corresponds in the scanning range on the target test paper strip through photoelectric sensor collection, according to the position of the rising edge of groove in the fluorescence curve, and the rising edge interval distance of predetermineeing the groove, can more accurately confirm the position of presetting the line, prevent to take place because of the inaccurate condition of presetting line position that test paper card holds in the palm the dislocation and leads to, confirm the relative peak value of fluorescence curve presetting line through the position of presetting line, and then can make the relative peak value of fluorescence curve presetting line that obtains more accurate, confirm the substance concentration of the sample that awaits measuring according to the relative peak value of fluorescence curve presetting line again, can make the accuracy of the substance concentration of the sample that awaits measuring that obtains higher.
Fig. 13 is a schematic structural diagram of a circuit device according to an embodiment of the present application, and referring to fig. 13, the circuit device includes: the memory 500 and the processor 600, wherein the memory 500 stores a computer program operable on the processor 600, and the processor 600 executes the computer program to implement the steps of the method for determining the concentration of the substance by the test strip.
In another aspect of the embodiments of the present application, a storage medium is further provided, where the storage medium stores a computer program, and when the computer program is executed by a processor, the computer program implements the steps of the method for determining a concentration of a substance by using a test strip.
In the embodiments provided in the present invention, it should be understood that the disclosed apparatus and method may be implemented in other ways. For example, the above-described apparatus embodiments are merely illustrative, and for example, a division of a unit is merely a logical division, and an actual implementation may have another division, for example, a plurality of units or components may be combined or integrated into another system, or some features may be omitted, or not executed. In addition, the shown or discussed mutual coupling or direct coupling or communication connection may be an indirect coupling or communication connection through some interfaces, devices or units, and may be in an electrical, mechanical or other form.
Units described as separate parts may or may not be physically separate, and parts displayed as units may or may not be physical units, may be located in one place, or may be distributed on a plurality of network units. Some or all of the units can be selected according to actual needs to achieve the purpose of the solution of the embodiment.
In addition, functional units in the embodiments of the present invention may be integrated into one processing unit, or each unit may exist alone physically, or two or more units are integrated into one unit. The integrated unit may be implemented in the form of hardware, or in the form of hardware plus a software functional unit.
The integrated unit implemented in the form of a software functional unit may be stored in a computer readable storage medium. The software functional unit is stored in a storage medium and includes several instructions to enable a computer device (which may be a personal computer, a server, or a network device) or a processor (in english: processor) to execute some steps of the methods according to the embodiments of the present invention. And the aforementioned storage medium includes: a U disk, a removable hard disk, a Read-Only Memory (ROM), a Random Access Memory (RAM), a magnetic disk or an optical disk, and other various media capable of storing program codes.
The above description is only for the specific embodiments of the present application, but the scope of the present application is not limited thereto, and any person skilled in the art can easily think of the changes or substitutions within the technical scope of the present application, and shall cover the scope of the present application. Therefore, the protection scope of the present application shall be subject to the protection scope of the claims.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (8)
1. A method for determining a concentration of a substance by a test strip, comprising:
acquiring a corresponding fluorescence curve in a scanning range on a target test strip through a photoelectric sensor;
according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove, the position of a preset line is determined, and the preset line comprises: a quality control line and a detection line;
determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line;
determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve;
the position of the preset line is determined according to the position of the rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset groove, and the method comprises the following steps:
preprocessing the fluorescence curve to obtain a first derivative curve of the fluorescence curve;
determining the position of a rising edge in the fluorescence curve according to the first derivative curve;
determining the position of the preset line according to the position of the rising edge and the distance between the rising edges of the test strip preset groove, wherein the distance between the rising edges of the test strip preset groove comprises: the test strip groove is a preset distance from the rising edge to the quality control line, and the test strip groove is a preset distance from the rising edge to the detection line;
determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line, wherein the determining comprises the following steps:
determining the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line according to the position of the preset line;
and determining the relative peak value of the preset line according to the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line.
2. The method of claim 1, wherein determining the fluorescence peak of the predetermined line and the fluorescence value of the comparison baseline of the predetermined line according to the position of the predetermined line comprises:
acquiring a fluorescence peak value of the preset line corresponding to the position of the preset line;
respectively determining baselines on two sides of the preset line according to the fluorescence peak value of the preset line and the distance between the preset baselines;
and selecting the corresponding base line with a higher fluorescence value from the base lines at the two sides of the preset line as a comparison base line, and recording the fluorescence value of the comparison base line.
3. The method of claim 1, wherein determining the corresponding peak value of the predetermined line according to the peak value of the predetermined line and the fluorescence value of the comparison baseline of the predetermined line comprises:
and taking the difference value of the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline as the relative peak value of the preset line.
4. The method of claim 1, wherein determining the substance concentration of the test sample from the relative peak values of the fluorescence curve comprises:
judging whether the detection result of the test strip is valid or not according to the fluorescence peak value of the quality control line;
and if the detection result is valid, determining the substance concentration of the sample to be detected according to the relative peak value of the detection line.
5. An apparatus for determining a concentration of a substance by a test strip, comprising: the device comprises an acquisition module, a position determination module, a peak value determination module and a concentration determination module;
the acquisition module is used for acquiring and acquiring a corresponding fluorescence curve in a scanning range on the target test strip through the photoelectric sensor;
the position determination module is used for determining the position of a preset line according to the position of a rising edge in the fluorescence curve and the distance between the rising edges of the test strip preset grooves, and the preset line comprises: a quality control line and a detection line;
the peak value determining module is used for determining the relative peak value of the preset line of the fluorescence curve according to the position of the preset line;
the concentration determination module is used for determining the substance concentration of the sample to be detected according to the relative peak value of the preset line of the fluorescence curve;
the position determining module is specifically configured to preprocess the fluorescence curve to obtain a first derivative curve of the fluorescence curve; determining the position of a rising edge in the fluorescence curve according to the first derivative curve; determining the position of the preset line according to the position of the rising edge and the distance between the rising edges of the test strip preset groove, wherein the distance between the rising edges of the test strip preset groove comprises: the test strip groove is a preset distance from the rising edge to the quality control line, and the test strip groove is a preset distance from the rising edge to the detection line;
the peak value determining module is specifically configured to determine a fluorescence peak value of the preset line and a fluorescence value of the comparison baseline of the preset line according to the position of the preset line; and determining the relative peak value of the preset line according to the fluorescence peak value of the preset line and the fluorescence value of the comparison baseline of the preset line.
6. A system for determining a concentration of a substance by a test strip, the system comprising: optical detection equipment, circuit equipment and test paper strips;
the circuit device employs the steps of the method of any one of claims 1 to 4, and the test strip is detected by the optical detection device.
7. A circuit device, comprising: memory in which a computer program is stored which is executable on the processor, and a processor which, when executing the computer program, carries out the steps of the method according to any one of the preceding claims 1 to 4.
8. A storage medium, characterized in that the storage medium has stored thereon a computer program which, when being executed by a processor, carries out the steps of the method of any one of claims 1 to 4.
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