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Method for extracting protein from feces

A feces and protein technology, applied in the preparation of test samples, etc., can solve problems such as interference test results, inconsistent protein content data, misdiagnosis or missed diagnosis of clinical diseases, etc., to achieve guaranteed purity, high protein output, and long centrifugation time Effect

Active Publication Date: 2015-10-28
THE FIFTH PEOPLES HOSPITAL OF SHANGHAI FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many protein extraction methods, but none of them are aimed at the extraction of protein in feces. The obtained protein content data is not uniform, which interferes with test results, and even leads to misdiagnosis or missed diagnosis of clinical diseases.

Method used

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  • Method for extracting protein from feces
  • Method for extracting protein from feces

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1 The method for extracting protein in feces (1)

[0020] The method for extracting protein in feces of the present invention comprises the following steps:

[0021] (1) Take columnar feces to avoid urine and water pollution, divide it into three sections, select the middle section to peel, collect stool samples and put them in special boxes, close the special boxes and store them in ice for 30 minutes;

[0022] (2) Then transfer the stool sample to a -80°C refrigerator for storage until use;

[0023] (3) Accurately weigh the stool sample and put it into a centrifuge tube, add RIPA lysis solution, the amount of RIPA lysis solution is 7.5 microliters of RIPA lysis solution per mg of stool sample, vortex the sample with a vortex machine for 1 minute, and then refrigerate Water bath for 10 minutes, repeat the steps of vortex and ice water bath 3 times until there are no visible fecal particles in the sample;

[0024] (4) Centrifuge the vortex liquid at 4°C and 1...

Embodiment 2

[0025] Example 2 Method for Extracting Protein in Feces (2)

[0026] The method for extracting protein in feces of the present invention comprises the following steps:

[0027] (1) Take columnar feces to avoid urine and water pollution, divide it into three sections, select the middle section to peel off, collect stool samples and put them in special boxes, close the special boxes and store them in ice for 25 minutes;

[0028] (2) Then transfer the stool sample to a -80°C refrigerator for storage until use;

[0029] (3) Accurately weigh the stool sample into a centrifuge tube, add RIPA lysis solution, the amount of RIPA lysis solution is 12.5 microliters of RIPA lysis solution per mg of stool sample, vortex the sample with a vortex machine for 1.5 minutes, and then freeze Water bath for 15 minutes, repeat the steps of vortex and ice water bath twice until there are no visible fecal particles in the sample;

[0030] (4) Centrifuge the vortex liquid at 4°C and 14,000g for 5 ...

Embodiment 3

[0031] Example 3 Method for Extracting Protein in Feces (3)

[0032] The method for extracting protein in feces of the present invention comprises the following steps:

[0033] (1) Take columnar feces to avoid urine and water pollution, divide it into three sections, select the middle section to peel, collect stool samples and put them in special boxes, close the special boxes and store them in ice for 35 minutes;

[0034] (2) Then transfer the stool sample to a -80°C refrigerator for storage until use;

[0035] (3) Accurately weigh the stool sample into a centrifuge tube, add RIPA lysate, the amount of RIPA lyse is 10 microliters of RIPA lyse per mg of stool sample, vortex the sample for 0.5 minutes with a vortex machine, and then freeze Water bath for 10 minutes, repeat the steps of vortex and ice water bath 4 times until there are no visible fecal particles in the sample;

[0036] (4) Centrifuge the vortex liquid at 4°C and 12,000g for 10 minutes in an ice-water bath, t...

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Abstract

The invention discloses a method for extracting protein from excrement. The method comprises the following steps of: dividing columnar excrement into three sections, peeling the middle section, collecting an excrement sample, putting the excrement sample into a special box, and putting the special box into ice for storage; transferring the excrement sample into a -80 DEG C refrigerator for storage and later use; accurately weighing the excrement sample, putting the excrement sample into a centrifugal tube, adding RIPA (radio immunoprecipitation assay) lysate, vortexing the sample by using a vortexing machine, carrying out ice-water bath, and repeating the steps of vortexing and ice-water bath until excrement particles which are macroscopic are completely removed; and performing ice-water bath on the vortex solution, then centrifuging at 4 DEG C, taking the supernatant fluid, centrifuging the supernatant fluid again, and preserving the final supernatant fluid in the -80 DEG C refrigerator. Each step of the method comprises ice bath, so that the possibility of degrading the protein is reduced, the protein is fully dissociated, and the protein yield is high; and the method for extracting protein is good in reproducibility, high in precision, and long in centrifugation time, ensures the purity of the protein, and is convenient to operate and easy to control.

Description

technical field [0001] The invention relates to the technical field of clinical stool examination, in particular to a method for extracting protein in stool. Background technique [0002] Stool examination is one of the routine clinical laboratory examination items. Through this examination, some pathological phenomena of the gastrointestinal tract can be intuitively understood, and the functional status of the digestive tract, pancreas, and liver and gallbladder can be judged indirectly. Diagnosis of tumors, checking for pathogenic bacteria in feces to prevent intestinal infectious diseases, etc. [0003] After the stool sample is collected, post-processing is usually carried out, followed by general trait inspection, formed component inspection, occult blood inspection, and inspection of bilirubin and its derivatives in stool. Among them, the protein content in feces is one of the laboratory inspection items. At present, there are many protein extraction methods, but non...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/42
Inventor 蔡元坤王会鹏
Owner THE FIFTH PEOPLES HOSPITAL OF SHANGHAI FUDAN UNIV