Urine preservation solution, preservation method and urine preservation tube

A technology for preserving liquid and urine, which is applied in chemical instruments and methods, preservation of human or animal bodies, and containers for laboratory use, etc. problems, to achieve high sensitivity, avoid the interference of cellular genomic DNA, and avoid the effect of microbial interference

Pending Publication Date: 2022-08-09
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complex composition of urine, it takes a certain time interval from sample collection to sample detection in clinical practice, and the cfDNA content in urine is low and easy to degrade, resulting in unsatisfactory detection results, such as hematuria patients and diabetic patients. The composition of urine is very complex

Method used

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  • Urine preservation solution, preservation method and urine preservation tube
  • Urine preservation solution, preservation method and urine preservation tube
  • Urine preservation solution, preservation method and urine preservation tube

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1. DNA Preservation Capacity

[0097] The preparation method of the urine preservation solution of the present embodiment: use ultrapure water to prepare 0.1wt% diethyl pyrocarbonate (DEPC) in a fume hood, use this DEPC solution as a solvent to prepare solution 1, and solution 1 contains the following mass percent ( wt%) final concentrations of reagents: 0.5% Tween 20, 6% ethylenediaminetetraacetic acid (EDTA), 5% glycine, 1.5% paraformaldehyde and 1.5% diazonium alkyl urea with hydrochloric acid and sodium hydroxide to adjust the pH of the solution to 8. The concentration of DEPC in solution 1 was estimated to be about 0.09 wt%. Use ultrapure water to prepare solutions 2, 3, and 4. Solutions 2, 3, and 4 respectively contain the final concentration reagents in mass percent: 44% Tris-HCl, 37% citric acid, and 67% trisodium citrate to prepare solutions In 2, 3, and 4, the pH of the solution was adjusted to 8 with hydrochloric acid and / or sodium hydroxide, respec...

Embodiment 2

[0103] Example 2. RNA preservation ability

[0104] In this case, the morning urine of a patient with hematuria (sample 4) was used. After mixing, each sample was divided into 8 parts, and each 10 mL part was placed in a nuclease-free centrifuge tube. The implementation group randomly selected 4 tubes according to the urine and the implementation case. 1 The prepared preservation solution was mixed at a ratio of 19:1 (this operation was completed within 0.5 hours of urine separation), the control group left the remaining 4 tubes of urine untreated, the implementation group (implementation case) and the control group (control The difference is that no urine preservation solution was added to the control group, and the two groups of specimens were stored at room temperature. One tube was taken from each of the two groups at the following 4 time points for whole urine and urine sediment cell RNA extraction and RNA QPCR (real-time quantitative PCR) detection. Store at room temper...

Embodiment 3

[0108] Example 3. Ability to preserve DNA and RNA simultaneously

[0109] In this case, morning urine of patients with hematuria (sample 5 and sample 6) was used. After mixing, each sample was divided into 8 parts, and each 10 mL part was put into sterile centrifuge tubes. The preservative solution prepared in Example 1 was mixed at a ratio of 19:1 (this operation was completed within 1 hour of urine ex vivo), and the remaining 4 tubes of urine in the comparison group were stored in a urine storage tube A (contrast reagent, commodity number: CW2657) and the preservation solution of urine preservation tube A (this operation is completed within 1 hour of urine in vitro). The difference between the implementation group (the example group) and the control group is that the preservation solution is different. save. At the following 4 time points, 1 tube was taken from each of the two groups for free nucleic acid extraction from the urine supernatant. The urine was stored within 2 ...

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Abstract

The invention discloses a urine preservation solution which is an aqueous solution containing the following substances in percentage by mass: 0.02%-0.1% of an RNA (Ribonucleic Acid) enzyme inhibitor, 2%-10% of a DNA (Deoxyribose Nucleic Acid) enzyme inhibitor, 0.2%-0.8% of a nonionic surfactant, 1%-5% of a cell fixing agent, 1%-5% of a preservative and 1%-5% of an aldehyde group quencher, the RNA enzyme inhibitor at least comprises diethyl pyrocarbonate, and the DNA enzyme inhibitor at least comprises ethylenediamine tetraacetic acid. The invention further discloses a urine preservation method and a urine preservation tube. After urine is in vitro and is preserved by adopting the urine preservation solution provided by the invention, relatively accurate free DNA and RNA information can still be provided, the degradation of nucleic acid substances is inhibited, interference signals including but not limited to genome DNA and microorganism interference are inhibited, and an overall detection solution result still has relatively high sensitivity and relatively high accuracy. The operation is simple and the shelf life is long.

Description

technical field [0001] The invention relates to the technical field of biological sample preservation, in particular to a urine preservation solution, a preservation method and a urine preservation tube. Background technique [0002] Urine is the terminal metabolite formed by the filtration and reabsorption of blood by the kidneys. Urine not only contains excretions in the body such as inorganic salts, uric acid, urea, glucose, etc., but also contains metabolites and genetic variations of other organs in the body. information, such as cell-free DNA (cfDNA), circulating tumor DNA (ctDNA), etc. Therefore, urine can be used as a biological sample for the detection and diagnosis of some urological diseases and other systemic diseases. For example, cfDNA in urine can be used for tumor detection. Early screening diagnosis. Urine collection is completely non-invasive and can be collected at home. It will be a very worthwhile specimen type for early screening and diagnosis of tumor...

Claims

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Application Information

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IPC IPC(8): C12N15/10A01N1/02B01L3/00
CPCC12N15/10A01N1/0226A01N1/0215A01N1/0221A01N1/021A01N1/0263A01N1/0268B01L3/50
Inventor 洪梅彭磊胡志红陈琪李金良戴立忠
Owner SANSURE BIOTECH INC
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