Excrement parameter and detection method and device thereof

Abstract

The excrement parameter is used for evaluating the number of target detection objects in excrement, and the excrement parameter is the number of the detection objects corresponding to unit dry matter; the quantity of the detection substances corresponding to the unit dry substances is equal to the quantity of the target detection substances in the excrement sample / the mass of the dry substances in the excrement sample; the mass of dry matter in the excrement sample is the mass of the excrement sample after drying and dewatering. An excrement parameter detection method is used for detecting the number of target detection objects in excrement. In the application, the applicant provides an excrement parameter for evaluating the amount of the target detection object in the excrement, and the excrement parameter is the amount of the detection object corresponding to the unit dry matter = the amount of the target detection object in the excrement sample / the mass of the dry matter in the excrement sample. The invention further provides an excrement parameter detection method and device which are used for obtaining the number of detection objects corresponding to unit dry matter. Based on the dry matter mass, a dimensional basis for excrement quantification is established, and accurate quantitative analysis excrement parameters can be obtained.

Description

Technical Field

[0001] This application belongs to the technical field of analysis of formed elements based on microscopic magnified images, specifically relates to the detection and analysis of formed elements in feces, and particularly relates to a fecal parameter for quantitative analysis of formed elements, as well as a detection method and device therefor. Background Art

[0002] During the fecal detection process, the target analytes to be detected include various formed elements such as residues, crystals, red blood cells, and white blood cells. However, the water content of feces is different in different species, different defecation times, and different physical conditions, and it is impossible to establish a quantitative evaluation standard for the target analytes.

[0003] In some studies, after collecting 24-hour fecal samples and mixing them, quantitative analysis was hoped to be carried out. However, in different species, different time periods, and different physical conditions, even for the collected 24-hour samples, due to the different water content and overall quality of the samples, it is impossible to find a quantitative benchmark.

[0004] The applicant has proposed a series of Chinese patents, such as

[0005] 1. CN2020112669290, "Cell Analysis Method and System and Quantitative Method and System";

[0006] 2. CN2020112669182, "Imaging Method and System for Cell Suspension Samples and Kit";

[0007] 3. CN2022104799126, "Fast Focusing Method for Microscopic Image Acquisition Device and Microscopic Image Acquisition Method";

[0008] 4. CN2023105340140, "Fecal Component Detection Method and System and Turbidimetry Dilution Card and Method";

[0009] 5. CN2023113529188, "Detection Method and Device for Formed Elements in Fecal Suspension";

[0010] 6. CN2023103592077, "Quantitative Analysis and Control Method and System for Fecal Formed Elements Based on Microscopic Images";

[0011] 7. CN2024100344351, "Fecal Suspension Parameter and Detection and Dry Matter Calculation Formula Obtaining Method and Device";

[0012] A brand-new technical solution is used to measure the content of target analytes in blood, urine, and feces, opening up a brand-new technical route for detecting formed elements in suspensions.

[0013] In the Chinese patent application "CN2023103592077", "Method and System for Quantitative Analysis and Control of Formed Components in Feces Based on Microscopic Images", the applicant proposed a method for detecting fecal dry matter and an evaluation system for quantitatively evaluating the content of formed components in feces. However, how to accurately measure the content of the target analyte in feces is a new technical challenge in the field of fecal measurement. Summary of the Invention

[0014] In this application, the inventor proposed a fecal parameter for evaluating the amount of the target analyte in feces. The fecal parameter is the number of analytes corresponding to unit dry matter = the number of target analytes in the fecal sample / the mass of dry matter in the fecal sample. The inventor also proposed a fecal parameter detection method and device for obtaining the number of analytes corresponding to unit dry matter. Based on the mass of dry matter, a dimensional basis for fecal quantification was established, enabling accurate quantitative analysis of fecal parameters.

[0015] The technical solution of this application to solve the above technical problems is a fecal parameter for evaluating the amount of the target analyte in feces. The above fecal parameter is the number of analytes corresponding to unit dry matter; the number of analytes corresponding to unit dry matter = the number of target analytes in the fecal sample / the mass of dry matter in the fecal sample; the mass of dry matter in the above fecal sample is the mass after the fecal sample is dried and dehydrated.

[0016] The above fecal sample is a fecal suspension sample obtained after dilution.

[0017] The above number of target analytes is obtained through microscopic imaging analysis; the dry matter of the above fecal suspension can also be obtained through the following method: from the image obtained by microscopic imaging, obtain the characteristic data of the formed components; calculate the mass of the dry matter of the above fecal suspension according to the characteristic data of the formed components and the dry matter calculation formula.

[0018] The above target analyte is any one of the following: intestinal protozoa, pathogenic microorganisms, parasite eggs, cells, food, food residues. Cells include red blood cells, white blood cells, and epithelial cells. Food residues include starch granules, plant fibers, or muscle fibers.

[0019] The technical solution of this application to solve the above technical problems can also be a fecal parameter detection method for detecting the amount of the target analyte in feces. The fecal sample is pretreated to obtain a fecal microscopic examination sample; the fecal microscopic examination sample is laid flat, and the height of the sample laying flat is H; take an image of the sample laying flat, and select the image corresponding to the area S1 of the sample laying flat. The detection volume V1 corresponding to the area S1 = S1 × H; identify the number of target analytes N in the detection volume V1 through the above image; the mass of dry matter corresponding to the volume V1 is M1; the number of analytes corresponding to unit dry matter = N / M1.

[0020] The above pretreatment includes: diluting the fecal sample to obtain a first-stage sample so that the target analyte and fecal impurities are dispersed; taking a part of the first-stage sample, enriching the target analyte, excluding fecal impurities, to obtain a second-stage sample, and the enrichment volume ratio is K; using the second-stage sample as the fecal microscopy sample.

[0021] Dilute the fecal sample with a reference turbidimetry card to obtain a first-stage sample.

[0022] In the above fecal parameter detection method, take the first-stage sample, measure the dry matter mass GMV corresponding to the unit volume; the detected dry matter mass M1 corresponding to the detection volume V1 = detection volume V1 × K × GM.

[0023] If the dry matter mass GMV corresponding to the above unit volume > 6 micrograms per microliter, or the dry matter mass GMV corresponding to the unit volume < 0.05 micrograms per microliter; it is determined that the preparation of the first-stage sample is unqualified.

[0024] By taking the first-stage sample, the sample volume is equal to V1 × K, and drying to obtain the detected dry matter mass M1.

[0025] The above fecal parameter detection method includes any one of the following technical features: Feature TA1: Enrich the target analyte in the first-stage sample, and the above enrichment is precipitation enrichment and / or centrifugation enrichment; after enrichment, take the suspension in the layer where the fecal target analyte is located as the second-stage sample; Feature TA2: Enrich the target analyte in the first-stage sample, add a flocculant to the first-stage sample, remove the impurities separated by flocculation, and retain the layer where the detection target is located to obtain the enriched second-stage sample.

[0026] The above target analyte is any one of the following: intestinal protozoa, pathogenic microorganisms, parasite eggs, cells, food, food residues. Cells include red blood cells, white blood cells, and epithelial cells. Food residues include starch granules, plant fibers or muscle fibers.

[0027] The above target analyte is lipid droplets, which float on the top of the fecal microscopy sample, and take a photo of the top of the fecal microscopy sample to obtain an image.

[0028] The above fecal sample pretreatment includes: diluting the fecal sample to obtain a fecal microscopy sample.

[0029] The above target analyte is pathogenic microorganisms.

[0030] The technical solution of the present application to solve the above technical problem can also be a fecal parameter detection device for detecting the amount of the target analyte in feces, based on the above fecal parameter detection method.

[0031] The technical effects of the above technical solution include: the number of analytes corresponding to unit dry matter is used to evaluate the content of the target analyte in feces based on dry matter; using the mass of unit dry matter as an objective measurement basis can well measure the content of bacteria and viruses in feces. It avoids the quantitative analysis deviation caused by the difference in water content in feces excreted by different species, at different times, and in different physical states; using the number of analytes corresponding to unit dry matter to evaluate the content level of the target analyte makes such parameters more valuable for clinical diagnosis with quantitative analysis.

[0032] The technical effects of the above technical solution include: for the image obtained by microscopy, the characteristic data of the formed components in the image can indirectly obtain the mass of dry matter through an empirical calculation formula, which brings convenience to the test.

[0033] The technical effects of the above technical solution include: based on the mass data of the formed components in the sample after turbidimetry, judge whether the sample preparation is qualified, so that the formed components in the fecal suspension sample after turbidimetry are ensured to be appropriate, and on this basis, there is a unified observation benchmark for further quantitative analysis. Ensure the consistency of subsequent quantitative analysis.

[0034] The technical effects of the above technical solution include: a dimensional basis for fecal quantification is established based on the mass of unit dry matter. Establish a unified basis for quantitative analysis of different target analytes, and the relative content of different target formed components in feces, that is, the target analyte relative to the mass of dry matter, can be obtained through calculation.

[0035] The technical effects of the above technical solution include: the method for obtaining the mass of dry matter can be obtained by identifying through the image method or by drying equal samples, and the two methods can verify each other, and expand the ways to obtain the mass of dry matter.

[0036] The technical effects of the above technical solution include: for the first-stage sample, enrich the target analyte to obtain the second-stage sample, and the enrichment volume ratio is K. Through enrichment, for low-concentration target analytes, the relative content of the target analyte relative to the mass of dry matter can be accurately obtained by means of existing methods.

[0037] The technical effects of the above technical solution include: after precipitation enrichment and / or centrifugation enrichment, take the suspension in the layer where the fecal target analyte is located as the fecal sample, and quantitative analysis can be carried out on various target analytes in different states, whether they are floating or sinking target analytes.

[0038] The technical effects of the above technical solution include: for the flocculant, remove the impurities separated by flocculation and retain the layer where the detection target is located to obtain the enriched fecal sample. The flocculant can remove impurities, improve the detection efficiency of the target analyte, and improve the recognition accuracy of the target analyte.

[0039] The technical effects of the above technical solutions include: the target detection object can be all the interested formed components in feces, such as intestinal protozoa, pathogenic microorganisms, parasite eggs, red blood cells, white blood cells; or the target detection object is starch granules, plant fibers or muscle fibers; or the target detection object is lipid droplets, and the lipid droplets float on the top of the detection sample, and an image is obtained by taking a picture of the top of the detection sample. Brief Description of the Drawings

[0040] Figure 1 is a schematic diagram of a method for detecting fecal parameters;

[0041] Figure 2 is a schematic diagram of a method for detecting fecal parameters;

[0042] Figure 3 is a schematic diagram of a method for detecting fecal parameters;

[0043] Figure 4 is a schematic diagram of a turbidimetry card used in a method for detecting fecal parameters;

[0044] Figure 5 is a schematic diagram of a method for detecting fecal parameters;

[0045] Figure 6 is a schematic diagram of taking multiple pictures of a microscopic examination sample. Detailed Embodiments

[0046] The following further details the content of the present application in conjunction with each drawing. It should be noted that the following is a description of the preferred embodiments of the present invention and does not constitute any limitation to the present invention. The description of the preferred embodiments of the present invention is only for the description of the general principles of the present invention. The numbers such as "first", "second", "A", and "B" involved in the present invention are only for the convenience of description and do not represent the order relationship in time or space. The combinations of letters and numbers "TA", "TB", and "H" involved in the present invention are only for the convenience of description, and the specific meanings are determined by the specific words they represent.

[0047] A fecal parameter is used to evaluate the amount of the target detection object in feces. The above fecal parameter is the number of detection objects corresponding to the unit dry matter; the number of detection objects corresponding to the unit dry matter = the number of target detection objects in the fecal sample / the dry matter mass in the fecal sample; the dry matter mass in the fecal sample is the mass after the fecal sample is dried and dehydrated.

[0048] In microscopic examination, diluting the fecal sample with water is required to prepare the fecal suspension. Regardless of the dilution factor, the dry matter in the fecal suspension corresponding to the same amount of sample remains unchanged. The dry matter in a sample does not vary with the dilution degree. Therefore, the content of the target analyte in feces, such as bacteria, viruses, cells, etc., can be evaluated based on the dry matter.

[0049] Although the water content in feces excreted by different species, at different times, and in different physical states is different, the content level of the target analyte can be evaluated by the number of analytes corresponding to the unit dry matter. For example, the content of Escherichia coli can be measured by the number of Escherichia coli corresponding to the unit dry matter to evaluate the Escherichia coli carriage level of an organism. By extension, the carriage levels of other intestinal bacteria can also be evaluated by the number of analytes corresponding to the unit dry matter, and such parameters have clinical diagnostic reference value.

[0050] The fecal sample can be a fecal suspension sample obtained after dilution. The number of target analytes is obtained through microscopic imaging analysis.

[0051] The dry matter of the fecal suspension can also be obtained by the following method: obtaining the characteristic data of the formed elements from the image of microscopic imaging; calculating the mass of the dry matter of the fecal suspension according to the characteristic data of the formed elements and the dry matter calculation formula.

[0052] Using the mass of the unit dry matter as an objective measurement basis can well measure the content of bacteria and viruses in feces. However, in fecal microscopy, the feces are always diluted to prepare a suspension and then microscopically examined, and the volume required for microscopy is small. The mass of the dry matter corresponding to this part of the volume (obtained by weighing) is very low and difficult to accurately measure, and the steps for obtaining it by drying are cumbersome. However, through the image obtained by microscopy, the characteristic data of the formed elements in the image can indirectly obtain the mass of the dry matter through an empirical calculation formula, which brings convenience to the test. After a large number of experiments and actual verification with high-precision weighing instruments, the accuracy of this method can meet the test accuracy requirements.

[0053] The target analyte is any one of the following: intestinal protozoa, pathogenic microorganisms, parasite eggs, red blood cells, white blood cells, starch granules, plant fibers, or muscle fibers.

[0054] Pathogenic microorganisms are small in volume and large in number, and their content can be accurately obtained from a relatively small volume of microscopic examination sample. During the microscopic examination process, the measurement of the number of pathogenic microorganisms and the mass of the dry matter can be carried out simultaneously.

[0055] Such as Figure 1, an embodiment of a fecal parameter detection method for detecting the amount of a target analyte in feces. The fecal sample is pre-treated to obtain a fecal microscopy sample; the fecal microscopy sample is laid flat, and the height of the flat sample is H; an image of the flat sample is taken, and the image corresponding to the selected area S1 of the flat sample is selected. The detection volume V1 corresponding to the area S1 = S1 × H; the number N of target analytes in the detection volume V1 is identified through the image; the dry matter mass M1 corresponding to the volume V1; the number of analytes per unit dry matter = N / M1.

[0056] Such as Figure 6 , S1 is the sum of the selected areas in multiple images. Of course, S1 can also be the selected area in one image. In the figure, the circular part is obtained by microscope optics, and on the image sensor, it is a square part. When taking multiple images, it is not necessary to align the edges of each image. For ease of calculation, the image corresponding to some pixels in the image can be selected.

[0057] Each selected image corresponds to a liquid volume. For example, it corresponds to an actual area with a length of 0.3 mm and a width of 0.2 mm, and the liquid height in this area is 0.4 mm. The volume corresponding to one image is 0.2 × 0.3 × 0.4 = 0.024 cubic millimeters. To increase the number of detected samples, multiple images are taken to obtain a larger sample volume. By taking 1000 images, a sample volume of 24 cubic millimeters can be obtained.

[0058] For a sample volume of 24 cubic millimeters, the dry matter mass in 24 cubic millimeters is greater than 0.05 × 24 = 1.2 micrograms and less than 6 × 24 = 144 micrograms. 144 micrograms requires precise measuring instruments for accurate measurement, and if measured after drying, it also requires cumbersome experimental steps. However, through the image feature data, it can be obtained from the dry matter calculation formula, which is simple and convenient.

[0059] The range of the dry matter mass per unit volume GMVV is: 0.05 micrograms per microliter ≤ GMV ≤ 6 micrograms per microliter. In this range, with a liquid height between 0.1 mm and 0.5 mm, there is better imaging quality.

[0060] Such as Figure 2 In [reference], the dry matter mass per unit volume can be obtained by taking images and calculating with an empirical formula; after obtaining the dry matter mass per unit volume, the dry matter mass can be calculated according to the volume of the test sample. How to obtain the dry matter mass per unit volume by taking images and calculating with an empirical formula is clearly described in another patent "CN2024100344351 Method and Device for Obtaining Fecal Suspension Parameters, Detection, and Dry Matter Calculation Formula" submitted by the applicant.

[0061] Such as Figure 3, the pretreatment includes: diluting the original fecal sample to obtain a first-stage sample, so that the target analyte and fecal impurities are dispersed; taking a part of the first-stage sample, enriching the target analyte and excluding fecal impurities to obtain a second-stage sample, and the enrichment volume ratio is K; using the second-stage sample as the fecal microscopy sample.

[0062] In the feces excreted by different species, at different times, and in different physical states, the content changes of intestinal protozoa, parasite eggs, red blood cells, white blood cells, starch granules, plant fibers or muscle fibers are large, and the number of analytes corresponding to the unit dry matter is one or more orders of magnitude different from the number of analytes corresponding to the unit dry matter of pathogenic microorganisms. When detecting these target analytes, sample enrichment is required to improve the detection accuracy.

[0063] To obtain the mass of dry matter corresponding to the unit volume through image shooting and empirical formula calculation, the feces need to be diluted to a certain extent. At the same time, there are a large number of impurities in the feces. Through the production of two-stage samples, the impurities can be greatly reduced and the target analytes can be enriched.

[0064] Such as Figure 4 , which is a turbidity reference diagram of a fecal suspension. Diluting the fecal sample with a reference turbidimetric card to obtain a first-stage sample can dilute the feces to a certain turbidity range, ensure that the impurity content in the microscopy image is within a relatively suitable range, and facilitate the observation of the microscopy target.

[0065] Such as Figure 3 , in an embodiment of a fecal parameter detection method, taking the first-stage sample and measuring the mass of dry matter corresponding to the unit volume GMV; the mass of detected dry matter M1 corresponding to the detection volume V1 = detection volume V1 × K × GMV. If the mass of dry matter corresponding to the unit volume GMV > 6 micrograms per microliter, or the mass of dry matter corresponding to the unit volume GMV < 0.05 micrograms per microliter; it is determined that the preparation of the first-stage sample is unqualified.

[0066] By measuring the mass of dry matter corresponding to the unit volume, the quality of sample preparation can be evaluated, and the unqualified sample quality caused by individual differences can be excluded.

[0067] Such as Figure 5 , the first-stage sample can be taken, the sample volume is equal to V1 × K, and dried to obtain the mass of detected dry matter M1.

[0068] Enrich the target analytes in the first-stage sample, and the enrichment is precipitation enrichment and / or centrifugation enrichment; after enrichment, take the suspension in the layer where the fecal target analyte is located as the second-stage sample.

[0069] For the detection item of parasite eggs, due to the large specific gravity of the eggs, precipitation or centrifugation has a good effect on egg enrichment.

[0070] It can also be to add a flocculant to the first-stage sample, remove the impurities separated by flocculation, and retain the layer where the detection target is located to obtain an enriched second-stage sample.

[0071] For some detection items with low specific gravity, such as red blood cells and white blood cells, the flocculant can quickly separate impurities. After separating the impurities, secondary precipitation and layering are carried out, which can effectively enrich the target analyte.

[0072] For detection items such as intestinal protozoa, pathogenic microorganisms, parasite eggs, red blood cells, white blood cells, starch granules, plant fibers or muscle fibers, after enrichment, the content in the second-stage sample can be increased by several orders of magnitude, and the detection accuracy is greatly improved.

[0073] If the target analyte is lipid droplets, the lipid droplets float on the top of the fecal microscopy sample, and an image is obtained by photographing the top of the fecal microscopy sample.

[0074] For pathogenic microorganisms with high content, enrichment may not be required, and a fecal sample can be directly diluted to obtain a fecal microscopy sample.

[0075] A fecal parameter detection device for evaluating the amount of a target analyte in feces, based on the above fecal parameter detection method.

[0076] Although the present invention is described and illustrated according to preferred embodiments and several alternative solutions, the invention is not limited by the specific descriptions in this specification. Other additional alternatives or equivalent components can also be used to practice the present invention.

Claims

1. A fecal parameter for evaluating the amount of a target analyte in feces, characterized in that the fecal parameter is the number of analytes corresponding to unit dry matter; the number of analytes corresponding to unit dry matter = the number of target analytes in the fecal sample / the mass of dry matter in the fecal sample; the mass of dry matter in the fecal sample is the mass after the fecal sample is dried and dehydrated.

2. The fecal parameter according to claim 1, wherein The fecal sample is a fecal suspension sample obtained after dilution.

3. The fecal parameter according to claim 2, wherein, The number of target analytes is obtained by microscopic imaging analysis; The dry matter of the fecal suspension can also be obtained by the following method; From the image obtained by microscopic imaging, characteristic data of formed elements are obtained; The mass of dry matter of the fecal suspension is calculated according to the characteristic data of formed elements and the dry matter calculation formula.

4. The fecal parameter according to any one of claims 1 to 3, characterized in that, The target analyte is any one of the following: intestinal protozoa, pathogenic microorganisms, parasite eggs, cells, food residues.

5. A fecal parameter detection method for detecting the amount of a target analyte in feces, characterized in that The fecal sample is pre-treated to obtain a fecal microscopic examination sample; The fecal microscopic examination sample is laid flat, and the height of the sample laying is H; An image of the laid-flat sample is taken, and the image corresponding to the area S1 of the laid-flat sample is selected. The detection volume V1 corresponding to the area S1 = S1 × H; The number of target analytes N in the detection volume V1 is identified through the image; The mass of dry matter corresponding to the volume V1 is M1; The number of analytes corresponding to unit dry matter = N / M1.

6. The fecal parameter detection method according to claim 5, characterized in that, The pre-treatment includes: Taking a fecal sample and diluting it to obtain a first-stage sample, so that the target analyte and fecal impurities are dispersed; Taking a part of the first-stage sample, enriching the target analyte, excluding fecal impurities, to obtain a second-stage sample, and the enrichment volume ratio is K; Using the second-stage sample as the fecal microscopic examination sample.

7. According to the method for detecting the mass of dry matter corresponding to unit volume described in claim 6, characterized in that The fecal sample is diluted with a reference turbidimetry card to obtain a first-stage sample.

8. According to the fecal parameter detection method described in claim 6, characterized in that Taking the first-stage sample, measuring the mass of dry matter corresponding to unit volume GMV; The detected dry matter mass M1 corresponding to the detection volume V1 = the detection volume V1 × K × GM.

9. According to the fecal parameter detection method described in claim 8, characterized in that If the mass of dry matter corresponding to unit volume GMV > 6 micrograms per microliter, or the mass of dry matter corresponding to unit volume GMV < 0.05 micrograms per microliter; it is determined that the preparation of the first-stage sample is unqualified.

10. According to the fecal parameter detection method described in claim 6: By taking the first-stage sample, the sample volume is equal to V1 × K, and drying to obtain the detected dry matter mass M1.

11. The fecal parameter detection method according to claim 6, wherein It includes any one of the following technical features: Feature TA1: Enriching the target analyte in the first-stage sample, and the enrichment is precipitation enrichment and / or centrifugation enrichment; after enrichment, taking the suspension in the layer where the fecal target analyte is located as the second-stage sample; Feature TA2: Enriching the target analyte in the first-stage sample, adding a flocculant to the first-stage sample, removing the separated impurities by flocculation, and retaining the layer where the detection target is located to obtain the enriched second-stage sample.

12. The fecal parameter detection method according to claim 6, characterized in that The target detection object is any one of the following: intestinal protozoa, pathogenic microorganisms, parasite eggs, cells, and food residues.

13. The fecal parameter detection method according to claim 6, characterized in that, The target detection object is a lipid droplet, and the lipid droplet floats on the top of the fecal microscopy sample, and an image is obtained by photographing the top of the fecal microscopy sample.

14. The fecal parameter detection method according to claim 5, wherein, The pretreatment of the fecal sample includes: diluting the fecal sample to obtain a fecal microscopy sample.

15. The fecal parameter detection method according to claim 14, wherein The target detection object is a pathogenic microorganism.

16. A fecal parameter detection device for detecting the amount of a target detection object in feces, characterized in that Based on the fecal parameter detection method according to any one of claims 5 to 15.

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