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Saliva protection agent

A protective agent and saliva technology, applied in the field of biology, can solve problems affecting the accuracy of analysis results, destroying DNA integrity, microbial contamination, etc., to achieve the effect of ensuring integrity and long-term stability, and broadening the range of protection conditions

Inactive Publication Date: 2015-11-11
SHANGHAI MAJORBIO BIO PHARM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DNA used for genetic testing should first ensure the purity and integrity. If the DNA is damaged such as breakage, it will directly affect subsequent tests such as PCR, thereby affecting the accuracy of the analysis results; in addition, saliva preservation fluid is used as a preservation medium for saliva DNA. , there is also a risk of oxidation and microbial contamination
These microorganisms may also promote the accelerated degradation of DNA and destroy the integrity of DNA. At the same time, after the saliva is contaminated by microorganisms, the DNA extracted from it will be impure, and eventually the integrity and purity of DNA cannot be guaranteed, which will lead to the failure of follow-up tests. Reliability is greatly reduced

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 saliva protectant preparation

[0030] According to the formula in the following table, dissolve each component in distilled water to prepare the saliva protectant, and adjust the pH value to 7.5-8.5. Among them, the reagents are all chemically pure.

[0031]

Embodiment 2

[0032] Embodiment 2 collects saliva sample and quality inspection

[0033] 1. Mix the collected saliva samples as samples;

[0034] 2. The sample and the saliva protectant prepared in Example 1 are mixed in a volume ratio of 1:1;

[0035] 3. Subpackage, label, and store under different conditions for later use;

[0036] 4. Extract preserved saliva DNA with tissue extraction kits at different time points;

[0037] 5. Use Nanodrop to check the concentration and purity of sample DNA, and electrophoresis to check the integrity of DNA.

[0038] 6. Target gene PCR amplification, PCR product sequencing detection.

Embodiment 3

[0039] Embodiment 3 preservation effect analysis

[0040] Sample information: Sample source, LQ and LSB are two saliva samples saved on April 15, 2014, each 7.5ml, numbered 140415LQ and 140415LS; another sample was donated by the company’s colleagues on April 21, 2014, after mixing Pack into 9 tubes, mark the date and number respectively: 14042101-14042109, the sample volume of each tube is 7.5ml; 14042101-14042103 are saliva samples mixed with water in equal proportions, and 14042104-14042106 are protective agents bought from the market (product Standard No.: YZB / Minxia 0022-2013) mixed with saliva samples for preservation, 14042107-14042109 is the protective agent prepared in Example 1 of the present invention and saliva equal ratio preservation. The test plan and implementation monitoring are shown in Table 1. At different time nodes as shown in the table below, the DNA of the saliva sample was extracted with the same tissue extraction kit, and quantified, electrophoresis ...

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Abstract

The invention relates to the field of biology, and in particular relates to a saliva protection agent as well as a preparation method and application thereof. Each 1000ml of the saliva protection agent consists of 10-50mmol of tris-hydroxymethyl aminomethane, 0.5-20mmol of ethylenediaminetetraacetic acid and / or sodium ethylene diamine tetracetate, 200-1000mmol of ammonium sulfate and 1-50mmol of glucose. Saliva, preserved in the saliva protection agent disclosed by the invention, is nearly free from change in the first 60 days, still complete in DNA and free from obvious degradation; the saliva is remained so after 6 months; and after 12 months, the saliva still remains the integrity of DNA as before. The saliva protection agent disclosed by the invention can be used for quite effectively preserving a saliva sample and for guaranteeing the integrity and long-term stability of the sample.

Description

technical field [0001] The invention relates to the field of biology, in particular to a saliva protectant and its preparation and application. Background technique [0002] In the field of biology, DNA extraction of blood and other body fluid samples plays an important role in the fields of molecular biology, genomics and medical testing. The DNA usually used for PCR amplification or detection is extracted from anticoagulated blood, but the blood is easy to agglomerate when stored for a long time, and the collection of blood will also cause certain trauma to the organism, and it requires professionals to collect blood safely. Therefore, the source of blood sample DNA limits its wide range of applications. [0003] Human saliva is a body fluid containing human cells. It is simple, convenient, efficient and non-invasive to collect. It is an ideal body fluid sample and suitable for large-scale sampling. However, the main components of saliva are water, oral mucosa cells, bac...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
Inventor 李胜彬于丹宋泽世胡秋萍张祥林
Owner SHANGHAI MAJORBIO BIO PHARM TECH
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