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Pretreatment method, preservation method, automatic treatment system and detection method of urine sample

A urine sample and storage method technology, applied in the field of automatic processing system and urine sample pretreatment, can solve the problems of not being easy to provide reproducibility and throughput, slow speed of pretreatment experiment process, many links, etc., to achieve Improve effectiveness and accuracy, reduce labor intensity, and improve the effect of adsorption rate

Pending Publication Date: 2022-04-15
JIANGSU QLIFE MEDICAL TECH GRP CO LTD
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  • Application Information

AI Technical Summary

Problems solved by technology

Among them, extracting protein from clinical samples, obtaining protein solution for content determination, reductive alkylation, proteolysis, peptide desalting and enrichment, the above processes all belong to the traditional manual sample processing stage in proteomics. Traditional manual pretreatment The experimental process is slow, has many links and is laborious. The whole process takes 8-18 hours. Usually, the experimenter cannot complete the pretreatment process of a batch of samples within one working day, and it is not easy to provide samples that meet the current clinical needs. High reproducibility and throughput, so high-throughput, automated proteomics pretreatment process is already one of the innovative technologies that are urgently needed in the industry

Method used

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  • Pretreatment method, preservation method, automatic treatment system and detection method of urine sample
  • Pretreatment method, preservation method, automatic treatment system and detection method of urine sample
  • Pretreatment method, preservation method, automatic treatment system and detection method of urine sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] A pretreatment method for a urine sample is roughly the same as that of Comparative Example 1, the difference lies in the concentration of the protein lysate, and the difference is as follows.

[0089] Sample B: Take 200 μL of the same urine sample, add 200 μL of 8M urea (diluent: 50 mM ammonium bicarbonate), the final concentration of urea is 4M, vortex and mix well, and extract protein.

Embodiment 2

[0091] A pretreatment method for a urine sample is roughly the same as that of Comparative Example 1, the difference lies in the concentration of the protein lysate, and the difference is as follows.

[0092] Sample C: Take 300 μL of the same urine sample, add 200 μL of 8M urea (diluent: 50 mM ammonium bicarbonate), the final concentration of urea is 3M, vortex and mix well, and extract protein.

Embodiment 3

[0094] A kind of preservation method of urine sample (referring to figure 1 ), which includes the following steps.

[0095] (1) Protein cleavage

[0096] Take 300 μL of the same urine sample (sample C), (diluent: 50 mM ammonium bicarbonate), the final concentration of urea is 3 M, vortex and mix well, and extract protein.

[0097] (2) Reductive alkylation

[0098] Add dithiothreitol to the product after protein cleavage to a final concentration of 10mM, and react at room temperature for 20min; add iodoacetamide (alkylation) to the product after reduction to a final concentration of 20mM, and react in the dark for 20min; An equal volume of dithiothreitol was added to the alkylated product to neutralize excess iodoacetamide in the alkylation reaction.

[0099] (3) Protein enrichment

[0100] Add 200 μL of 70% ethanol to the PVDF filter plate and centrifuge at 1000g to activate the PVDF filter plate; Equilibrate; then transfer the sample to a PVDF filter plate and centrifuge...

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Abstract

The invention discloses a pretreatment method, a preservation method, an automatic treatment system and a detection method of a urine sample, and relates to the technical field of biological detection.The pretreatment method comprises the steps that the urine sample subjected to protein splitting is subjected to reductive alkylation treatment, and then protein enrichment and enzymolysis are conducted; in the protein enrichment step, a PVDF (Polyvinylidene Fluoride) filter plate is adopted to carry out protein enrichment on the sample subjected to reductive alkylation treatment. The invention further provides an automatic urine sample treatment system and an automatic sample treatment method. The treatment system greatly reduces the labor intensity of people, facilitates the improvement of the treatment efficiency of urine sample treatment, meets the high-flux and automatic proteomics pretreatment requirements, and adapts to the reproducibility and flux of the current clinical requirements.

Description

technical field [0001] The present invention relates to the technical field of biological detection, in particular to a pretreatment method, preservation method, automatic processing system and detection method of urine samples. Background technique [0002] Changes in the type and quantity of proteins in urine carry a variety of information on the occurrence, development and prognosis of urinary system diseases, and urine proteomics is one of the most effective methods to interpret the information carried by urine proteins. In the field of clinical applications, multi-center and large samples are often required for research and analysis in order to obtain more accurate results. Due to the large volume of urine samples, large differences in protein concentration, and the presence of microbial characteristics such as bacteria, how to optimize the preservation of clinical urine samples is one of the problems that need to be solved urgently. [0003] Urine samples have the adv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/40G01N1/42G01N1/28G01N30/02G01N30/06G01N30/08G01N30/34G01N30/72G01N35/10
CPCG01N35/10C12Y105/03004C12N9/0032C12N9/6427C12Y304/21004G01N1/4044G01N1/34G01N1/42G01N30/34G01N30/72G01N2001/4088
Inventor 成晓亮余静周岳张伟郑可嘉
Owner JIANGSU QLIFE MEDICAL TECH GRP CO LTD
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