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 contamin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0029] Example 1 Preparation of saliva protectant

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

[0031]

Example Embodiment

[0032] Example 2 Collection of saliva samples and quality inspection

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

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

[0035] 3. Divide packaging and labeling, save for use under different conditions;

[0036] 4. Extract the preserved saliva DNA with tissue extraction kit at different time nodes;

[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.

Example Embodiment

[0039] Example 3 Analysis of preservation effect

[0040] Sample information: sample source, LQ and LSB are saliva samples of two people saved on April 15, 2014, 7.5ml each, numbered 140415LQ and 140415LS; in addition, samples were donated by company colleagues on April 21, 2014, after mixing Pack 9 tubes, marked with the date and number: 14042101—14042109, each tube has a sample volume of 7.5ml; 14042101-14042103 is a mixture of saliva samples and water in equal proportions, and 14042104-14042106 is a protective agent (product Standard number: YZB / Minxia 0022-2013) and saliva samples were mixed and stored, 14042107-14042109 were the protective agent prepared in Example 1 of the present invention and saliva were stored in equal proportions. The test plan and implementation monitoring are shown in Table 1. At different time nodes in the following table, use the same tissue extraction kit to extract DNA from saliva samples, and quantify, electrophoresis quality inspection, partial...

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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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