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Cell preserving fluid and using method thereof

A technology for preserving liquid and cells, applied in the field of biomedicine, can solve the problems of limiting nucleic acid detection capacity, manpower and material consumption, etc., and achieve the effect of saving sample collection costs, speeding up the detection process, and speeding up the efficiency of nucleic acid detection.

Inactive Publication Date: 2021-04-27
SUZHOU VERSABIO TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the sensitivity of nucleic acid detection is about 30-50%. In order to avoid false negatives, at least two tests are required to confirm the diagnosis;

Method used

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  • Cell preserving fluid and using method thereof
  • Cell preserving fluid and using method thereof
  • Cell preserving fluid and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Use 1% sodium lauryl sulfate, 0.1% guanidine isothiocyanate, 0.1% ethylenediaminetetraacetic acid, 0.05% sodium chloride, 0.05% potassium chloride, 0.05% disodium hydrogen phosphate, 0.05% dihydrogen phosphate Potassium, 0.05% Tris-HCl buffer solution (pH=8.0), 0.1% sucrose and 25ug / ml Bianka penicillin were prepared as cell preservation solution. For samples of fresh HeLa cell line placed in cell preservation solution and stored at 37°C for 0 day, 1 day, 3 days, and 7 days, after RNA was extracted using Qiagen’s RNeasy Mini kit, fluorescent quantitative PCR was performed to detect ACTB, GADPH , IL1B, TLR4 and other four genes, each group of samples was repeated 3 times. like figure 1 As shown, fresh RNA and RNA treated with cell preservation solution had no significant change in the RNA detection Ct values ​​of the four genes within 0-7 days of storage at 37°C.

Embodiment 2

[0024] Use 1% guanidine isothiocyanate, 0.1% ethylenediaminetetraacetic acid, 0.05% sodium chloride, 0.05% potassium chloride, 0.05% disodium hydrogen phosphate, 0.05% potassium dihydrogen phosphate, 0.01% Tris-HCl buffer (pH=8.0), 0.1% glucose and 25ug / ml bianka penicillin were prepared as a cell preservation solution. Using the high-risk HPV in vitro molecular diagnostic kit of Suzhou Guoke Wenpu Biotechnology Co., Ltd., the fresh HeLa cell line and the cell preservation solution were stored at 4°C and room temperature (RT) for 0 days, 7 days, 14 days, and 21 days. For the samples of Day 1 and Day 28, the DNA was extracted using the Genome Extraction Kit of Suzhou Guoke Wenpu Biotechnology Co., Ltd., and then detected by fluorescent quantitative PCR. Each group of samples was repeated 3 times.

[0025] like figure 2 As shown, there is no significant difference in Ct value between fresh DNA and DNA treated with cell preservation solution at 4°C and room temperature for 28 d...

Embodiment 3

[0027] Use 1% sodium lauryl sulfate, 0.1% guanidine isothiocyanate, 0.1% ethylenediaminetetraacetic acid, 0.05% sodium chloride, 0.05% potassium chloride, 0.05% disodium hydrogen phosphate, 0.05% dihydrogen phosphate Potassium, 0.01% Tris-HCl buffer solution (pH=8.0), 0.1% sucrose and 25ug / ml biankapenicillin were prepared as a cell preservation solution. For samples of fresh HeLa cell lines placed in cell preservation solution and stored at 37°C for 0 day, 1 day, 3 days, and 7 days, the alcohol-based CytoPrep preservation solution of VersaBio Company and the Pierce preservation solution of Thermo Fisher Corporation were used at the same time Preserve fresh HeLa cell lines in cell preservation solution and store at 37°C for 0 days, 1 day, 3 days, and 7 days. After the RNA was extracted by Qiagen’s RNeasy Mini kit, four genes including ACTB, GADPH, IL1B, and TLR4 were detected by fluorescent quantitative PCR, and each group of samples was repeated 3 times. like image 3 As sh...

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Abstract

The invention belongs to the field of biomedicine, and particularly relates to a cell preserving solution and a use method thereof, the preserving solution contains 0.1%-10% of a nucleic acid stabilizer, 0.05%-1% of an osmotic pressure stabilizer, 0.01%-0.1% of a pH stabilizer, 0.05%-1% of a freeze-drying protective additive and the balance of water. The cell preserving fluid provided by the invention has better time stability and temperature stability, and can achieve the effects of saving the sample collection cost and accelerating the nucleic acid detection efficiency for regions which cannot be delivered to a detection site on the same day and are lack of effective transportation equipment.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a cell preservation solution and a use method thereof. Background technique [0002] The single-stranded RNA virus, which is both the gold standard for diagnosis and the gold standard for general screening, has poor RNA stability, especially at room temperature and for long-term storage. Specimens involving virus or nucleic acid detection should be stored in a 4°C refrigerator for no more than 24 hours. It can be seen that the sample collection preservation solution has a great influence on the sensitivity of the subsequent detection results. At present, the sensitivity of nucleic acid detection is about 30-50%. In order to avoid false negatives, at least two tests are required to confirm the diagnosis; therefore, the ability of nucleic acid detection is limited to a certain extent, and the consumption of manpower and material resources is extremely serious. For areas tha...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/021
Inventor 赵国栋熊尚岷
Owner SUZHOU VERSABIO TECH INC
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