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Saliva preserving fluid and preparation method thereof

A technology for preserving liquid and saliva, applied in the field of molecular biology, can solve problems such as bacterial growth and unfavorable DNA integrity, and achieve the effects of protecting integrity, protecting cell stability and reducing preparation costs.

Inactive Publication Date: 2021-03-05
武汉远赞吉诺百客医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the sucrose component in it can maintain the osmotic pressure, it will cause the growth of bacteria and microorganisms; although the guanidine isothiocyanate component can denature the protein in saliva, it will lyse the cells and leave the DNA in a free state, which is not conducive to maintaining DNA integrity

Method used

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  • Saliva preserving fluid and preparation method thereof
  • Saliva preserving fluid and preparation method thereof
  • Saliva preserving fluid and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0032] (1) Preparation of saliva preservation solution

[0033] S1, prepare EDTA with a concentration of 0.1mol / L, SDS with a mass volume fraction of 10%, and PBS buffer solution with a pH of 7.4. The PBS buffer solution is prepared according to volume percentage: 72% NaCl solution, 1% KCl solution, 2 HPO 4 12H 2 O solution 25% and KH 2 PO 4 Solution 2%;

[0034] S2, sterilizing the EDTA and PBS buffer solution at 100°C for 10 minutes;

[0035] S3, after cooling the sterilized EDTA and PBS buffer to 20°C, mix according to the following ratio: EDTA5ml, SDS5ml, Tween 200.5ml, preservative 0.05ml, add PBS buffer to 50ml;

[0036] S4, filter the mixed solution in step S3 with a 0.20 μm filter membrane, and store it at 20° C. after the preparation is completed.

[0037] Among them, the PBS buffer, calculated as 1L of PBS buffer, includes the following components: 100mmol / L of NaCl, 1.5mmol / L of KCl, 7mmol / L of Na 2 HPO 4 12H 2 O and 1 mmol / L of KH 2 PO 4 , the solvent is...

Embodiment 2

[0043] (1) Preparation of saliva preservation solution

[0044] S1, respectively prepare EDTA with a concentration of 0.8mol / L, SDS with a mass volume fraction of 15%, and PBS buffer solution with a pH=7.4. The PBS buffer solution is prepared according to volume percentage: 75% NaCl solution, 2% KCl solution, 2 HPO 4 12H 2 O solution 20% and KH 2 PO 4 Solution 3%;

[0045] S2, sterilizing the EDTA and PBS buffer solution at 130°C for 30 minutes;

[0046] S3, after cooling the sterilized EDTA and PBS buffer to 30°C, mix according to the following ratio: EDTA15ml, SDS15ml, Tween201.5ml, preservative 0.35ml, add PBS buffer to 50ml;

[0047] S4, filter the mixed solution in step S3 with a 0.30 μm filter membrane, and store it at 30° C. after the preparation is completed.

[0048] Among them, the PBS buffer, calculated as 1L of PBS buffer, includes the following components: 110mmol / L of NaCl, 1.8mmol / L of KCl, 9mmol / L of Na 2 HPO 4 12H 2 O and 1.3mmol / L of KH 2 PO 4 , th...

Embodiment 3

[0054] (1) Preparation of saliva preservation solution

[0055] S1, respectively prepare EDTA with a concentration of 0.3mol / L, SDS with a mass volume fraction of 13%, and PBS buffer solution with a pH=7.4. The PBS buffer solution is prepared according to volume percentage: 69.5% NaCl solution, 1.5% KCl solution, and 2 HPO 4 12H 2 O solution 24% and KH 2 PO 4 Solution 5%;

[0056] S2, sterilize the EDTA and PBS buffer solution at 110°C for 20 minutes;

[0057] S3, after cooling the sterilized EDTA and PBS buffer to 25°C, mix according to the following proportions: EDTA10ml, SDS12ml, Tween200.8ml, preservative 0.12ml, add PBS buffer to 50ml;

[0058] S4, filter the mixed solution in step S3 with a 0.25 μm filter membrane, and store it at 25° C. after the preparation is completed.

[0059] Among them, the PBS buffer, calculated as 1L of PBS buffer, includes the following components: 150mmol / L of NaCl, 2.5mmol / L of KCl, 10mmol / L of Na 2 HPO 4 12H 2 O and 2mmol / L of KH 2...

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Abstract

The invention provides a saliva preserving fluid and a preparation method thereof. The saliva preserving fluid comprises a cell stabilizer, a nuclease inhibitor, a surfactant, a protein denaturing agent and a preservative. The saliva preserving fluid has good broad-spectrum antibacterial activity, can inhibit pollution of bacteria, fungi and the like, enables genome DNA with good integrity and purity to still be obtained after a saliva sample has been preserved for one month at normal temperature, and can be applied to preservation of saliva of a subject in scientific researches such as gene detection and epidemiological investigation.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a saliva preservation solution and a preparation method thereof. Background technique [0002] In recent years, saliva sampling has gradually been adopted in the fields of scientific research and medicine to obtain individual DNA. For routine scientific research, saliva samples have obvious advantages over blood samples. amount of genomic DNA. Saliva mainly contains water, oral mucosal cells, bacteria, salivary amylase, mucin, glycoprotein, and inorganic substances. The DNA in saliva mainly comes from oral epithelial mucosal cells and salivary gland leukocytes. During the preservation of saliva, it is necessary to Maintain the integrity of DNA, inhibit the degradation of DNA by nucleases, and prevent bacteria, fungi and other microorganisms from polluting the purity of DNA. Therefore how to make saliva can also obtain DNA with higher purity under long-term preservation becomes ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0226
Inventor 易吉李泽卿郑婷婷
Owner 武汉远赞吉诺百客医学科技有限公司
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