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Blood cfDNA extracting kit

A kit and blood technology, applied in the biological field, can solve problems such as pollution, failure of cfDNA extraction, inaccurate test results, etc.

Inactive Publication Date: 2019-05-14
NINGBO AJCORE BIOSCIENCES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This new device helps protect against damage caused by bacteria or other harmful substances from affecting genetic material extracted through samples collected on plasma membranes (PCM). It uses an enclosed container called a preserve tube that keeps the sample safe before being used with PCR analysis techniques like qRT-PCNA.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the accuracy and reliability of detecting specific types or amounts of target polynucleotides present during different stages of disease development due to their ability to form stable duplex structures when subjected to external stimuli like heat stress or chemical agents.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] This embodiment discloses a blood cfDNA extraction kit, which includes a sample DNA storage tube, a lysate, a binding solution, and a washing solution, and the sample DNA storage tube is filled with a blood cfDNA storage solution. For the blood cfDNA preservation solution in the blood cfDNA extraction kit of the present invention, the applicant focused on the selection of new types of anticoagulant and membrane protection agent for research. Based on the unique chelating properties provided by multiple valence bonds in EDDHA and good compatibility with other components in blood / reagents, the applicant replaced the commonly used anticoagulant EDTA-3K with part of EDDHA–Na; based on the effect of rhamnose and trehalose on cells Good tonicity and surfactant properties under low toxicity and high salt conditions, choose sorbitol, trehalose, rhamnose as membrane protective agent.

[0022] According to the different anticoagulants used, the formulations are divided into three...

Embodiment 3

[0031] Embodiment 3: each formulation performance comparison

[0032] Test product: formula 1-14 blood sample DNA preservation tube obtained in Example 2;

[0033] Control substance: Kangjie lithium heparin blood collection tube: produced by Jiangsu Kangjie; Streck non-invasive vacuum blood collection tube: produced by Streck (unknown anticoagulant, protective agent, presumably EDTA and sugar, etc.); blank glass blood collection tube;

[0034] Most of the reagents used in the research stage are well-known imported reagents. Those skilled in the art can understand that these reagents can be replaced by similar / higher quality domestic / imported reagents for reasons of price / convenience, and achieve the same or similar effects.

[0035] Recruit 3 male volunteers aged 22-25 (after physical examination: normal blood routine indicators, no physical examination tumor indications, no inflammation indications), blood collection 180ml (formulation 1-14 blood collection tubes, Kangjiehepa...

Embodiment 4

[0046] Embodiment 4: the actual detection application of preferred formula

[0047] The free blood DNA storage tubes of formula 3 and formula 12 of this application were used together with Streck non-invasive vacuum blood collection tubes for the actual detection of EGFR T790M in tumor patients (cfDNA was extracted by QIAamp Circulating Nucleic Acid Kit kit, detected by cobas® EGFR Mutation Test kit, The operation is according to the standard procedure of the kit), involving a total of 42 patients. Each patient uses three kinds of blood collection tubes for sampling. The first test is performed within 48 hours after blood collection, and two tests are performed after 14 and 21 days of storage at room temperature to compare the test results.

[0048] Table 6 EGFR T790M detection results

[0049]

[0050] Except that the test results of the three storage methods within 48 hours and on the 14th in the formula 3 group were consistent, there were 3 cases of inconsistency (false ...

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Abstract

The invention relates to a blood cfDNA extracting kit. The kit comprises a sample DNA preserving tube, a lysis solution, a binding solution and a washing solution. The kit is characterized in that thesample DNA preserving tube is filled with a blood cfDNA preserving solution, and the preserving solution is prepared from 10-50 mg/ml of glucose, 1-5 mg/ml of glutamine, 1-5 mg/ml of glycine, 200-1,000 mg/ml of sodium orthovanadate, 1-5 mg/ml of genistein, 1-5 mg/ml of trasylol, 1-5 mg/ml of aluminum fluoride, 10-40 mg/ml of glutathione, 100-500 mg/ml of diazolidinyl urea, 1-3 mg/ml of an anticoagulant and 5-20 mg/ml of a membrane protection agent. According to the blood cfDNA extracting kit, the preserving tube for a blood sample is additionally arranged; the preserving solution guarantees the content of free DNA in plasma on the condition that preserved cells are not damaged, and the cell viability can be still kept high even after the sample is preserved for 21 days at normal temperature.

Description

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Claims

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Application Information

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Owner NINGBO AJCORE BIOSCIENCES INC
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