CN113916814A - Urine sample processing and detecting method - Google Patents

Urine sample processing and detecting method Download PDF

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CN113916814A
CN113916814A CN202111188203.4A CN202111188203A CN113916814A CN 113916814 A CN113916814 A CN 113916814A CN 202111188203 A CN202111188203 A CN 202111188203A CN 113916814 A CN113916814 A CN 113916814A
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urine
detecting
urine sample
value
samples
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邱永丰
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N2021/0106—General arrangement of respective parts
    • G01N2021/0112—Apparatus in one mechanical, optical or electronic block

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Abstract

The invention discloses a method for processing and detecting a urine sample, which is characterized by comprising the following steps of: step 1, collecting and detecting samples: collecting urine of the person to be tested, and detecting the absorbance value OD of the urine0(ii) a Step 2, processing and detecting samples: using a microfiltration membrane to process a urine sample to be detected, and detecting the absorbance value OD of the urine again1(ii) a And 3, numerical calculation and analysis: using the absorbance value OD0And the absorbance value OD1And obtaining a target value to analyze the urinary calculus stone forming risk tendency of the urine sample. If the target value is less than 1, the urine sample has a stone forming danger tendency, and the smaller the target value is, the larger the stone forming danger tendency is possibly; if the target value is not less than 1, the result is negative, and the urine sample has no stone forming danger tendency. The invention has the advantages of simple and convenient detection, low cost and wide application range of the method, and opens up a new way for researching the urinary calculus.

Description

Urine sample processing and detecting method
Technical Field
The invention relates to the field of urine detection, in particular to a method for processing and detecting a urine sample.
Background
Urinary calculus is one of the most common urological diseases in China, south China is one of three high incidence areas in the world, calculus can relapse in about 50% of patients within 10 years, and a simple, convenient and effective urinary calculus urine examination method is not used for analyzing the dangerous tendency of urinary calculus, even evaluating the curative effect of a medicament for treating calculus and the like.
At present, most of the methods for detecting the urinary calculus are methods for measuring oxalic acid and citric acid, namely ion chromatography, but the methods have inherent defects. Ion chromatography is a technology 20 years ago in principle, and the concentration of a substance in a liquid is measured by detecting an electric signal generated by ion exchange in the liquid by a detector. Due to inherent defects of electric signal detection in liquid, although the ion chromatography is high in theoretical precision, the ion chromatography has the defects of unstable signals, large errors of measurement results and the like in the actual operation process, and sometimes the measurement errors can reach more than 30% in the actual process. Particularly for the determination of cations, the peak value of the cation is small, and the area under the peak value is relatively small, so that the peak value of the cation is fused with other peaks at many times, and needs to be separated out through graphic processing, thereby undoubtedly increasing the error of the measurement of the cation. Firstly, the organic small molecules cannot be effectively measured due to large errors by using a colorimetric method; secondly, the cation measurement results can meet the clinical requirements, but still have the risk of large errors and even misdiagnosis.
Disclosure of Invention
In order to overcome the defect that the method for detecting the calculus in the prior art has larger error, the invention provides a method for treating and detecting a urine sample.
The invention is realized by adopting the following technical scheme:
a method for processing and detecting a urine sample comprises the following steps:
step 1, collecting and detecting samples: collecting urine of the person to be tested, and detecting the absorbance value OD of the urine0；
Step 2, processing and detecting samples: using a microfiltration membrane to process a urine sample to be detected, and detecting the absorbance value OD of the urine again1；
And 3, numerical calculation and analysis: using the absorbance value OD0And the absorbance value OD1And obtaining a target value to analyze the urinary calculus stone forming risk tendency of the urine sample.
Preferably, in step 1, the urine sample is processed and detected in 1 hour after fresh urine is excreted.
Preferably, in step 1, the urine sample is collected by a collection tube with a cover, and then placed on a multifunctional shaker for 1 minute at 20rpm, and then sampled for detection.
Preferably, in the step 2, the microfiltration membrane processor is a 0.22 μm water system needle filter, and the detection wavelengths of the microplate readers are 405nm,450nm and 490nm respectively.
Preferably, in the step 3, the target value is analyzed by SPSS13.0 statistical software.
Preferably, in the process of analyzing by using the SPSS13.0 statistical software, the detection results of 30 groups of urine samples are analyzed, the absorbance values before and after microfiltration are analyzed by using two groups of paired t-tests, and whether the results of the filtering results of the risk factors for calculus formation in the urine samples by the microfiltration membrane are statistically different is judged by judging whether the p value is less than 0.05.
The invention has the beneficial effects that:
the invention has the advantages of simple and convenient detection, low cost and wide application range of the method, and opens up a new way for researching the urinary calculus.
The key point of the invention is that the urine is detected by a microfiltration combined spectrophotometry (the spectrophotometry is not limited to a microplate reader), the urine is directly detected by the invention before and after microfiltration treatment, and the urine lithogenesis danger tendency is analyzed by taking the ratio of the two as a target value. The method is a research method for directly detecting urine by using a microfiltration combined spectrophotometry (the spectrophotometry is not limited to a microplate reader) and analyzing the dangerous tendency of urine forming stones by taking the ratio of the absorbance value of the urine after microfiltration treatment to the absorbance value without treatment as a target value.
Other alternatives may also be employed in the solution of the invention:
(1) ultrafiltration and nanofiltration are also membrane separation techniques and should be an alternative to the microfiltration process used in the present invention.
(2) Spectrophotometry has other detection devices besides microplate readers.
Detailed Description
For the purpose of more clearly illustrating the present invention and more clearly understanding the technical features, objects and advantages of the present invention, the technical solutions of the present invention will now be described in detail below, but are not to be construed as limiting the implementable scope of the present invention.
Microfiltration is the earliest membrane technology developed and applied in the world, microporous filter membranes are prepared in laboratories as early as 100 years ago, and with the development of polymer materials, the research on membrane forming mechanism and membrane preparing technologyThe progress of the operation makes the micro-filtration membrane rapidly developed. The micro-filtration membrane has the aperture of 0.01-10um generally, no medium falls off during filtration, no secondary pollution is caused, and the micro-filtration membrane is mainly used for intercepting particles from gas or liquid phase suspension and is widely applied to the aspects of food processing sterilization, juice clarification, pure draught beer filtration, tea beverage treatment, traditional Chinese medicine extraction, sewage treatment and the like. In 1852 Beer proposed the basic law of spectrophotometry (lambert-Beer law), in 1854 Duboscq and Nessler designed a first colorimeter based on this theory to be applied to the field of quantitative analytical chemistry, at present spectrophotometers are widely used in the fields of industrial and agricultural production and scientific research, can be used for the analysis of inorganic and organic compounds, and are suitable for the measurement of trace components, and the lower limit of measurable concentration is 10-5-10-6mol/L, and high accuracy, and the relative error is 1-5%. The microplate reader used in the research is also called a microplate detector, is actually a phase-change special spectrophotometer, and generally requires that the final volume of the test solution is below 250 uL. The difference in energy before and after the light passes through the object to be detected, that is, the energy absorbed by the object to be detected, the concentration of the same object to be detected and the absorbed energy at a specific wavelength are quantitatively related, the degree of absorption of the light by the substance is represented by absorbance, and the detection unit is represented by OD (optical density).
The urinary calculus formation is premised on the supersaturation of calculus salts in urine, and the calculus formation may undergo a series of dynamic processes such as urine supersaturation, nucleation, growth and aggregation, solid phase transfer and the like. Urinary calculus urine often contains high concentrations of stone-forming substances, which are risk factors for urinary calculus formation.
The invention is further described below with reference to the following examples.
Examples
A method for processing and detecting a urine sample comprises the following steps:
step 1, collecting a sample:
collecting urine of a person to be tested, collecting the urine by a collecting pipe with a cover within 1 hour after the fresh urine is discharged, placing the collecting pipe on a multifunctional oscillator, oscillating the collecting pipe for 1 minute at 20rpm, and extracting 5ml of urine by a 5ml syringe to place the urine into a small sterile test tube as a urine sample before treatment;
step 2, processing the sample:
after a 5ml syringe is used for extracting 5ml of urine, a microfiltration membrane processor (namely a 0.22 mu m water system needle filter) is connected to filter the urine and the urine enters a small test tube to be used as a processed urine sample;
step 3, detecting samples and calculating numerical values:
respectively sampling the urine sample before and after treatment, detecting with enzyme labeling instrument, and respectively detecting with wavelengths (λ) of 405nm,450nm, and 490nm to obtain absorbance value OD before treatment0And the absorbance value OD after sample treatment1Calculating to obtain a target value, wherein the target value is OD after processing1OD before treatment0。
In this embodiment, 10 samples are detected with each of three wavelengths (λ)405nm, 450nm, and 490nm, and 30 samples are detected, wherein 30 samples are positive, 0 samples are negative, and the positive rate is 100%.
Specific values are shown in tables 1-3 below.
Table 1 shows the results of detection at a wavelength (. lamda.) of 405nm, Table 2 shows the results of detection at a wavelength (. lamda.) of 450nm, and Table 3 shows the results of detection at a wavelength (. lamda.) of 490 nm. Wherein, the processed sample is the absorbance value OD of the sample processed by a 0.22 mu m microfiltration membrane1Before treatment, the absorbance value OD of the sample without 0.22 μm microfiltration membrane treatment0。
TABLE 1 detection results at a wavelength of 405nm
Figure BDA0003300149120000031
Figure BDA0003300149120000041
TABLE 2 detection results at a wavelength of 450nm
Figure BDA0003300149120000042
TABLE 3 detection results at 490nm wavelength
Figure BDA0003300149120000043
Step 4, numerical analysis:
using absorbance values (OD)1) And absorbance value (OD)0) The urinary calculus stone forming risk tendency of the urine sample is analyzed by taking the ratio of (1) as a target value, if the target value is less than 1, the urine sample has the stone forming risk tendency, and the smaller the target value is, the larger the stone forming risk tendency is possibly; if the target value is not less than 1, the result is negative, and the urine sample has no stone forming danger tendency. In addition, if the risk factors for calculus formation in the urine are filtered by the microfiltration membrane, the OD value of the urine before and after filtration changes, and the larger the change is, the more the stone-forming substance particles are filtered, the larger the stone-forming risk tendency is.
The invention has the advantages of simple and convenient detection, low cost and wide application range of the method, and opens up a new way for researching the urinary calculus.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims (6)

1. A method for processing and detecting a urine sample is characterized by comprising the following steps:
step 1, collecting and detecting samples: collecting urine of the person to be tested, and detecting the absorbance value OD of the urine0；
Step 2, processing and detecting samples: using a microfiltration membrane to process a urine sample to be detected, and detecting the absorbance value OD of the urine again1；
And 3, numerical calculation and analysis: using the absorbance value OD0And the absorbance value OD1And obtaining a target value to analyze the urinary calculus stone forming risk tendency of the urine sample.
2. The method as claimed in claim 1, wherein the urine specimen is processed and tested in 1 hour after fresh urine is excreted in step 1.
3. The method as claimed in claim 1, wherein in step 1, the urine specimen is collected by a collection tube with a cover, and then put on a multifunctional shaker for 1 minute at 20rpm, and then sampled for testing.
4. The method as claimed in claim 1, wherein in step 2, the microfiltration membrane processor is a 0.22 μm aqueous needle filter, and the detection wavelengths of the microplate reader are 405nm,450nm and 490nm, respectively.
5. The method as claimed in claim 1, wherein in step 3, the target value is analyzed by SPSS13.0 statistical software.
6. The method as claimed in claim 1, wherein the SPSS13.0 statistical software analyzes the detection results of 30 groups of urine samples, analyzes the absorbance values before and after microfiltration by two pairs of t-tests, and determines whether the p value is less than 0.05, thereby determining whether the risk factors of calculus formation in the urine samples are filtered out by the microfiltration membrane and the results are statistically different.
CN202111188203.4A 2021-10-12 2021-10-12 Urine sample processing and detecting method Pending CN113916814A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1118068A (en) * 1994-03-04 1996-03-06 株式会社京都第一科学 Method of and apparatus for measuring uric components
CN104764826A (en) * 2015-04-13 2015-07-08 中国烟草总公司四川省公司 Method for determining content of cotinine in urine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1118068A (en) * 1994-03-04 1996-03-06 株式会社京都第一科学 Method of and apparatus for measuring uric components
CN104764826A (en) * 2015-04-13 2015-07-08 中国烟草总公司四川省公司 Method for determining content of cotinine in urine

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
孙倩 等: "多功能离子液体分散液液微萃取结合高效液相色谱法检测人尿中 5 种邻苯二甲酸酯代谢物", 色谱, vol. 978, no. 8, pages 929 - 936 *
潘祖光 等: "吸光度下降法测定人尿中尿酸的含量", 右江民族医学院学报, vol. 17, no. 3, pages 260 - 261 *

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