Oral swab preservation solution as well as preparation method and application thereof

A technology of preservation solution and swab, which is applied in application, preservation of human or animal bodies, animal husbandry, etc., and can solve problems such as unfavorable preservation of DNA stability and integrity, unfavorable preservation of DNA integrity, unfavorable stability of genetic material, etc. , to achieve the effect of convenient storage and transportation, no toxic and side effects, and broad application prospects

Inactive Publication Date: 2019-04-30
CHEERLAND BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Existing patents are all saliva preservation liquid, which is consistent with the principle of oral swab preservation liquid, and its protective agent has complex components, such as patent publication numbers CN102440234B and CN102919218B
Wherein CN102919218B contains magnesium chloride, and magnesium ion is a nuclease activator, which easily causes the degradation of DNA, which is unfavorable for the preservation of the stability and integrity of DNA; CN102440234B contains protein denaturants such as guanidine isothiocyanate, which can inhibit nuclease activity, but at the same time, it also lyses the cells, leaving the DNA in a free state, which is not conducive to the integrity preservation of DNA
Containing sucrose and glucose components respectively in CN102919218B and CN105039306A, although can well keep the viscosity of this saliva preservation immobilization solution and maintain cell osmotic pressure, but easily cause the growth of microorganisms such as bacteria; The composition is complex in CN105368812A, although the composition such as urea, ethanol Organic substances such as mucin and globulin in saliva can be denatured, but at the same time, cells will be lysed, which is not conducive to the integrity of DNA, and these components provided in this application document are all necessary components, namely All components are indispensable to maintain the stability of DNA
[0008] In short, the existing technology has the following problems. The cryopreservation method can effectively inhibit the growth of microorganisms, but repeated freezing and thawing of oral epithelial cells can easily lead to cell lysis, which is not conducive to maintaining the stability of genetic material
Although normal saline preservation can maintain the stability of oral epithelial cell morphology, long-term storage is prone to the growth of bacteria, fungi and other microorganisms

Method used

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  • Oral swab preservation solution as well as preparation method and application thereof
  • Oral swab preservation solution as well as preparation method and application thereof
  • Oral swab preservation solution as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of embodiment 1 oral cavity swab preservation solution

[0043] The buccal swab preservation solution of this embodiment includes the following components and contents: Tris-HCl 35mmol / L, EDTA 3.2mmol / L, NaCl 150mmol / L, SDS 0.5% (w / v), and the system pH is 8.

[0044] The preparation method of the saliva preservation solution of the present embodiment is as follows:

[0045] (1) Measure 3.5mL Tris-HCl (1mol / L), 0.32mL EDTA (0.5mol / L), 15mL NaCl (1mol / L) and 5mL SDS (10%), and add them to a 100mL beaker in turn (beaker Clean with pure water), stir evenly with a clean glass rod, or mix evenly with a magnetic stirrer;

[0046] (2) adjust the pH value of the solution to 8.0 with dilute hydrochloric acid;

[0047] (3) Transfer the mixed solution in the beaker to a 100mL volumetric flask, and dilute to the mark with ultrapure water;

[0048] (4) Filter the above preservation solution with a disposable filter membrane device, store it in a storage bottle, an...

Embodiment 2

[0050] Example 2 Oral swab sample collection and preservation

[0051] In this embodiment, oral cavity cell collection and storage methods are as follows:

[0052] (1) Rinse your mouth 30 minutes before sampling to remove food residues and residual microorganisms. Do not eat, drink, brush your teeth, smoke or chew gum within 30 minutes after rinsing to avoid contamination of food DNA, and do not brush your teeth or drink a lot of water to avoid affecting The recovered number of exfoliated cells.

[0053] (2) Tear off the outer packaging of the oral swab, and carefully take out the oral swab.

[0054] (3) Hold the handle, insert the swab into the left oral cavity, make the head of the swab fully touch the inside of the left cheek (subject to the slight convexity of the cheek), and rub it up and down more than 20 times with the force of brushing your teeth.

[0055] (4) After sampling, take out the 2mL sampling tube (500 μL of preservation solution inside) in the packaging box...

Embodiment 3

[0058] Example 3 Buccal swab sample DNA extraction

[0059] In the present embodiment, sampling Tiangenzhu extraction method oral swab genomic DNA extraction kit extraction, the method is as follows:

[0060] (1) Take a sampling tube with 500 μL of preservation solution and oral swab in the tube, add 20 μL of proteinase K solution, vortex for 10 sec, and place at 56°C for 60 min, during which time, vortex for several times every 15 min.

[0061] (2) Add 400 μL buffer GB, mix thoroughly by inversion, and place at 70°C for 10 minutes. The solution should now be clear, centrifuge briefly to remove droplets from the inside of the cap, then squeeze the swab to transfer as much lysate as possible to a new centrifuge tube.

[0062] (3) Add 200 μL of absolute ethanol, mix thoroughly by inversion, and briefly centrifuge to remove the liquid droplets on the inner wall of the tube cap.

[0063] (4) Add the solution and flocculent precipitate obtained in the previous step into an adsorp...

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Abstract

An oral swab preservation solution comprises the following components: 5-50 mmol / L of Tris-HCl, 1-10 mmol / L of EDTA, 50 to 500 mmol / L of sodium chloride and 0.1 to 3% of SDS, wherein the pH range of the system ranges from 7 to 10. The oral swab preservation solution s clear in component, simple in preparation method, low in cost, good in stability and high in safety, free of toxic and side effects, easy to operate, convenient to store and transport, suitable for large-scale sampling and use, and wide in application prospect. The oral swab preservation solution is used for storing a human mouthswab sample, can be preserved at the temperature of 15-25 DEG C (room temperature.) for a long term, genomes in oral epithelial cells can be effectively protected from being damaged in a plurality ofweeks, the integrity of DNA (deoxyribonucleic acid) is good, and the oral swab sample is not polluted by microorganisms.

Description

technical field [0001] The invention relates to the technical field of cell preservation, in particular to a buccal swab preservation solution and its preparation method and application. Background technique [0002] DNA extraction is the most basic application technology in molecular biology testing. Traditionally, human whole blood is mostly used as the sample for gene extraction. There are certain difficulties in the sampling process, such as the damage of blood to the human body and the safety hazards in the process of blood collection. , Blood samples are easily contaminated, and the patient rejection rate is high. At the same time, the cost of taking blood samples is high, and the operation is relatively cumbersome. It requires professionally skilled personnel to operate it reliably. Therefore, it is of great significance to explore gene collection methods that are easier to obtain materials, more convenient to operate, and easier to be accepted by patients. [0003] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0226
Inventor 郭娟陈川陈建国王瑢杨传春张文勇
Owner CHEERLAND BIOTECH CO LTD
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