Liquid for eliminating microbial pollution and application method thereof

A microbial contamination and microbial technology, applied in the field of biochemistry, achieves the effects of simple preparation, easy promotion, and low production cost

Inactive Publication Date: 2019-02-05
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a liquid for removing microbial contamination, so as to easily and quickly remove the microbial contamination caused by storage of biological samples, cell culture, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Removal of Bacterial Contamination in SP2 / 0 Myeloma Cells

[0017] 1. Preparation of liquid to remove microbial contamination: take povidone (Aladdin reagent) and PEG 8000, add ultrapure water to dissolve, adjust pH to 7.0, constant volume, filter with 0.22 μm filter membrane to remove bacteria and impurities, spare.

[0018] 2. Add 1 / 2 volume of the above liquid to the contaminated SP2 / 0 myeloma cells, place in a 5% carbon dioxide incubator at 37°C for 2 hours; remove the supernatant, add 1640 medium to wash the cells once; remove Liquid, just add complete medium to continue culturing cells.

[0019] 3. Table 1 is the actual effect of using povidone-iodine alone as a fungicide to act on polluted SP2 / 0 cells. When the povidone-iodine concentration is 0.03% (special note: all percentages involved in the present invention are mass percentages unless otherwise specified), the bactericidal effect is moderate, and the cell activity is the best.

[0020] The impa...

Embodiment 2293

[0026] Example 2 The removal of 293T cell fungal contamination

[0027] 1. Preparation of liquid for removing microbial contamination: Take povidone and PEG 8000, add ultrapure water to dissolve, adjust pH to 8.0, add different volumes of DMSO (dimethyl sulfoxide), constant volume, and use 0.22 μm Membrane filter to sterilize and remove impurities; spare.

[0028] 2. Add 1 / 2 volume of the above liquid to the contaminated 293T cells, place in a 5% carbon dioxide incubator at 37°C for 12 hours; remove the supernatant, add PBS (phosphate buffer saline, pH=7.4) to wash Cells once; remove the liquid and add complete medium to continue culturing the cells.

[0029] 3. The experimental results are shown in Table 3, adding DMSO (3%-5%) can improve the bactericidal effect. But continue to increase the concentration of DMSO, the cell activity is inhibited.

[0030] Table 3 Adding DMSO to kill fungi effect evaluation

[0031] DMSO concentration (%, V / V)

Embodiment 3

[0032] Example 3 The removal of mycoplasma contamination in primary mouse fibroblasts

[0033] 1. Preparation of liquid for removing microbial contamination: Take povidone and PEG 8000, add ultrapure water to dissolve, adjust pH to 7.0, add magnesium sulfate, dissolve to constant volume, filter with 0.22μm filter membrane to remove bacteria and impurities . spare.

[0034] 2. Add 1 / 2 volume of the above liquid to the contaminated mouse primary fibroblasts, place in a 5% carbon dioxide incubator at 37°C for 5 hours; remove the supernatant, add PBS (phosphate buffered saline, pH7.4) Wash the cells once; remove the liquid, add complete medium to continue culturing the cells.

[0035] 3. The experimental results are shown in Table 4. Adding magnesium sulfate at a concentration of 15mM-25mM (mM means millimole per liter) can improve the bactericidal effect.

[0036] Table 4 Adding Magnesium Sulfate to Kill Fungi Efficacy Evaluation

[0037] Magnesium sulfate concentrat...

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PUM

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Abstract

The invention discloses liquid for eliminating microbial pollution and an application method thereof. The liquid contains povidone iodine and PEG 8000. The liquid capable of eliminating the microbialpollution provided by the invention can kill fungi, bacteria, viruses, mycoplasma and other microbial pollution in a broad-spectrum mode and has no obvious toxic effect on cells; and in addition, theliquid is simple to prepare, the preparation cost is low, the popularization is easy, and meanwhile, the problems of microbial pollution caused by biological sample storage, cell culture and the likecan be easily, conveniently and rapidly solved.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a liquid for removing microbial contamination and a use method. Background technique [0002] Povidone-iodine (PVP-I) is a complex of polyvinylpyrrolidone (PVP) and iodine, which not only retains the strong bactericidal properties of iodine, has no resistance to bacteria and viruses, but also overcomes the difficulty of dissolving free iodine in water , unstable, irritating to the skin and yellow staining and other shortcomings. This intermolecular complexation effectively reduces the corrosiveness and irritation of iodine. Iodine in povidone iodine solution exists as complex iodine with low toxicity, and 80%-90% of the complex iodine can be depolymerized into free iodine to exert bactericidal effect. It has a broad-spectrum killing effect on negative and positive bacteria, fungi and viruses. It is suitable for washing hands before surgery, cleaning and disinfection of surgical fiel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N59/12A01N25/02A01N25/22A01N59/06A01P1/00A01P3/00
CPCA01N25/02A01N25/22A01N59/06A01N59/12
Inventor 李金峰柯跃斌肖云军吕子全何洁
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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