CN103710338B - DNA extraction kit in a kind of human whole blood white corpuscle - Google Patents
DNA extraction kit in a kind of human whole blood white corpuscle Download PDFInfo
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Abstract
The invention provides DNA extraction kit in a kind of human whole blood white corpuscle, described test kit comprises erythrocyte cracked liquid and DNA extraction liquid, and described erythrocyte cracked liquid comprises following component: the sodium-chlor of 4.5 ~ 5.5mmol/L, 0.8 ~ 1.2%(v/v) Triton X-100, the glucose of 300 ~ 340mmol/L and the Tutofusin tris of 0.008 ~ 0.012mol/L.The invention solves the defect of existing whole blood nucleic acid extraction kit, provide a kind of operate fast simple, extraction efficiency is high, DNA rapid extraction test kit in the human whole blood white corpuscle of use safety, low cost of manufacture.Apply this test kit, rapid extraction can be carried out to DNA in the clinical whole blood sample white corpuscle detected for round pcr, extract the DNA of white corpuscle DNA and the white corpuscle inner virus obtained in whole blood.
Description
Technical field
The present invention relates to DNA extraction kit in a kind of human whole blood white corpuscle.
Background technology
From round pcr since last century, the eighties was come out, it is more and more extensive in the application of molecular biology association area.Especially in recent years, along with the raising that people require clinical diagnosis precision sensitivity, and the proposition of Personalized medicine concept and popularization, round pcr relies on its advantage such as quick, responsive, special in clinical diagnosis, be applied to clinic diagnosis widely rapidly, by detecting the gene in clinical samples, thus doctor is helped to determine medication and treatment plan.
And sample DNA extraction is the prerequisite that round pcr uses, human peripheral blood plays an important role in immune response and metabolism in human body, can be used for the molecular monitoring of blood system and other numerous systemic diseases, because it samples simple and easy to get clinically, be most widely used in clinical diagnosis or research.
Current whole blood DNA extracting method mainly contains phenol chloroform method, centrifugal column method, paramagnetic particle method.Wherein phenol chloroform method complicated operation and containing the rare use of organic solvent of large toxicity; Centrifugal column method remains current most popular whole blood DNA extracting method, but it needs special pellosil chromatography column, and could obtain DNA by wash-out after needing eccentric cleaning repeatedly; Paramagnetic particle method adopts magnetic bead absorption, cleaning wash-out obtains DNA; Need repeatedly centrifugal, replace tubes, bar magnet sepn process in these method operating process, complicated operation, extraction time is longer, higher to operational requirement, is unfavorable for clinic diagnosis or research application.
The existing multiple test kit for the extraction of whole blood gene can be applied to clinical both at home and abroad at present.The whole blood DNA extracting method that these test kits provide has a lot of shortcoming: as DNA extraction process is complicated, sample process length consuming time; During process, there is consume in various degree in the DNA in sample, especially for high density sample, cracking is insufficient, and enrichment is incomplete, causes DNA lose and make sample quantitatively on the low side; Treatment step is many, and repeatedly, bar magnet sepn process, easily causes crossed contamination and environmental aerosols between sample to pollute for the replace tubes that needs repeatedly to uncap, centrifugal, rinsing.Leaching process needs multiple instrument or special equipment, and manufacturing cost is higher, high to operational requirement.
Summary of the invention
The object of this invention is to provide a kind of human whole blood DNA rapid extraction test kit and from human whole blood white corpuscle, extract the method for DNA, solve the defect of existing whole blood nucleic acid extraction kit, provide a kind of operate fast simple, extraction efficiency is high, DNA rapid extraction test kit in the human whole blood white corpuscle of use safety, low cost of manufacture.Apply this test kit, rapid extraction can be carried out to DNA in the clinical whole blood sample white corpuscle detected for round pcr.
Therefore, the invention provides DNA extraction kit in a kind of human whole blood white corpuscle, described test kit comprises erythrocyte cracked liquid and DNA extraction liquid, and described erythrocyte cracked liquid comprises following component: the sodium-chlor of 4.5 ~ 5.5mmol/L, 0.8 ~ 1.2%(v/v) Triton X-100, the glucose of 300 ~ 340mmol/L and the Tutofusin tris of 0.008 ~ 0.012mol/L.
Preferably, in described erythrocyte cracked liquid, the concentration of described sodium-chlor is 5mmol/L, the concentration of described Triton X-100 is 1.0%(v/v), the concentration of described glucose is 320mmol/L, and the concentration of described Tutofusin tris is 0.01mol/L, surplus is sterilized water, and use sodium hydroxide or hydrochloric acid soln adjust ph to make the pH value of described erythrocyte cracked liquid be 7.8 ~ 8.6, preferably 8.0 ~ 8.4, more preferably 8.2.
In a kind of embodiment of the present invention, described DNA extraction liquid comprises the Repone K of 0.6 ~ 1.0mol/L, the ethylenediamine tetraacetic acid (EDTA) of 0.4 ~ 0.8mol/L, 0.8 ~ 1.2%(m/v) sodium lauryl sulphate, surplus is sterilized water, and use sodium hydroxide or hydrochloric acid soln adjust ph to make the pH value of described DNA extraction liquid be 7.8 ~ 8.6, preferably 8.0 ~ 8.4, more preferably 8.2.
In another kind of embodiment of the present invention, described DNA extraction liquid comprises Buddhist Sha graceful (surfactin) 0.01 ~ 0.5mmol/L, Repone K 20 ~ 300mmol/L, sodium laurylsulfonate 0.01 ~ 2% and ethanol 0.05 ~ 1%.
The present invention also provides a kind of using method of test kit described above, comprises the following steps, steps A, remove red corpuscle: in whole blood, add erythrocyte cracked liquid, fully shake mixing, obtain mixture, centrifugation is carried out to it, removes supernatant, leave white depositions; Step B, extracts DNA in white corpuscle: in steps A, add described DNA extraction liquid in gained white depositions, with pipettor piping and druming mixing, leaves standstill and makes the abundant cracking of white corpuscle and the DNA that discharges wherein, obtain the DNA solution for pcr amplification.
In the above-mentioned methods, preferably, repeating step A twice; Namely again erythrocyte cracked liquid is added to the white depositions in steps A, fully shake mixing, make precipitation Eddy diffusion and obtain mixed solution, centrifugation is carried out to it, removing supernatant, leaving white depositions.
Preferably, in steps A, volume of whole blood is more than 100 microlitres, and the volume ratio of whole blood and erythrocyte cracked liquid is 1:1 ~ 5.The rotating speed of centrifugation in preferred steps A is 10000 ~ 13000 revs/min, and the time of centrifugation is 1 ~ 3 minute.
Preferably, in steps A, in whole blood and step B, the volume ratio of DNA extraction liquid is 1:0.25 ~ 1.
The present invention also provides the application of a kind of test kit as above in fluorescent PCR detection technique.
Containing unique erythrocyte cracked liquid in test kit of the present invention, it makes the erythrocyte-specific lysis in whole blood sample, isolates the karyocyte in whole blood; The DNA of white corpuscle DNA in whole blood and white corpuscle inner virus is obtained again under the effect of DNA extraction liquid.Test kit in the application of the invention, greatly can avoid the loss of DNA in leaching process, considerably increase the extraction efficiency of whole blood DNA; The present invention uses less operation steps, reduces crossed contamination and the pollution to environment between pipe; Extraction step of the present invention is simple, without the need to heating, only needs to carry out centrifugal segregation supernatant liquor once or twice, then adds DNA extraction liquid, can complete whole extraction, substantially reduce extraction time, be conducive to the raising of clinical diagnosis working efficiency.Test kit of the present invention is cheap for manufacturing cost, without the need to special material or instrument, only needs whizzer and pipettor, use safety.The present invention can suppress Dnase and Rnase active simultaneously, and does not have an impact to subsequent experimental.Extraction efficiency of the present invention is higher than common whole blood DNA extracting method.
Embodiment
Embodiment 1
1, the compound method of DNA extraction kit and middle reagent thereof in human whole blood white corpuscle:
1. the preparation of erythrocyte cracked liquid: first add a small amount of sterilized water in volumetric flask, take and add the sodium-chlor making concentration finally for 5mmol/L, add the Triton solution that volume ratio is finally 1%, add the glucose that ultimate density is 320mmol/L, add the Tutofusin tris that ultimate density is 0.01mol/L; Adding sterilized water is settled to volume required, fully mixes dissolving, uses sodium hydroxide or hydrochloric acid soln adjust ph to 8.2; Autoclaving 10 minutes.
2. the preparation of DNA extraction liquid: first add a small amount of sterilized water in volumetric flask, take and add the Repone K making concentration finally for 0.8mol/L, add the ethylenediamine tetraacetic acid (EDTA) that ultimate density is 0.6mol/L, add the sodium lauryl sulphate (mass volume ratio) that ultimate density is 1%; Adding sterilized water is settled to volume required, fully mixes dissolving, uses sodium hydroxide or hydrochloric acid soln adjust ph to 8.2; Autoclaving 15 minutes.
2, the application of test kit:
The application of test kit is 100 routine whole blood samples.The concrete operation step of application is: in every example 200 microlitre whole blood, add 800 microlitre erythrocyte cracked liquids, and fully concussion mixing 10 seconds, obtains the limpid mixture of red, transparent; Carry out centrifugation to said mixture, centrifugal rotational speed is 12000 revs/min, and the centrifugation time is 1 minute, removes supernatant, stays white depositions; In above-mentioned throw out, add 800 microlitre erythrocyte cracked liquids again, fully shake mixing, make precipitation Eddy diffusion and obtain mixed solution; Carry out centrifugation to above-mentioned mixed solution, centrifugal rotational speed is 12000 revs/min, and the centrifugation time is 1 minute, removes supernatant, stays white depositions; In described throw out, add 100 microlitre DNA extraction liquid, with pipettor piping and druming mixing several, leave standstill 10 minutes, make the abundant cracking of white corpuscle, discharge DNA, obtain the DNA solution that can be used for pcr amplification.
3, to effect detection and the evaluation of test kit application:
β-Actin house-keeping gene detection reagent is used to detect above-mentioned DNA solution.As a result, the test kit in employing the present invention detects β-Actin house-keeping gene wherein after extracting 100 routine whole blood sample DNA, 100 examples all have amplification curve, illustrate that test kit provided by the invention can effectively extract and obtain leukocytic DNA in whole blood sample.
In addition, test kit of the present invention and commercially centrifugal column method of the prior art are extracted test kit and are carried out DNA extraction to same whole blood sample, show by β-Actin house-keeping gene detection kit detected result, to same 100 routine samples carry out process the present invention comparatively centrifugal column method save 2 hours, and the forward 1-4 of the detected result Ct extracted, visible test kit of the present invention is easier compared with the operation of centrifugal column method, and has more high extracting efficiency.
Embodiment 2
1, the compound method of DNA extraction kit and middle reagent thereof in human whole blood white corpuscle:
1. the preparation of erythrocyte cracked liquid: with embodiment 1.
2. the preparation of DNA extraction liquid: first add a small amount of sterilized water in volumetric flask, take and add the Repone K making concentration finally for 0.1mol/L, add Buddhist Sha graceful (surfactin) that ultimate density is 0.2mmol/L, to add ultimate density be the sodium laurylsulfonate (mass volume ratio) of 0.5% and add the ethanol that ultimate density is 0.2%; Adding sterilized water is settled to volume required, fully mixes dissolving, uses sodium hydroxide or hydrochloric acid soln adjust ph to 8.2; Autoclaving 15 minutes.
2, the application of test kit:
The application of test kit is 1 × 10 for containing epstein barr virus dna concentration
4the whole blood sample of IU.The concrete operation step of application is: in 200 microlitre whole bloods, add 800 microlitre erythrocyte cracked liquids, and fully concussion mixing 10 seconds, obtains the limpid mixture of red, transparent; Carry out centrifugation to said mixture, centrifugal rotational speed is 12000 revs/min, and the centrifugation time is 1 minute, removes supernatant, stays white depositions; In above-mentioned throw out, add 800 microlitre erythrocyte cracked liquids again, fully shake mixing, make precipitation Eddy diffusion and obtain mixed solution; Carry out centrifugation to above-mentioned mixed solution, centrifugal rotational speed is 12000 revs/min, and the centrifugation time is 1 minute, removes supernatant, stays white depositions; In described throw out, add 50 microlitre DNA extraction liquid, with pipettor piping and druming mixing several, leave standstill 10 minutes, make the abundant cracking of white corpuscle, discharge DNA, obtain the DNA solution that can be used for pcr amplification.
3, to effect detection and the evaluation of test kit application:
Epstein-Barr virus nucleic acid quantitative determination reagent kit is commercially used to detect above-mentioned DNA solution.As a result, it is 1 × 10 that the test kit in employing the present invention repeats to extract the concentration detected wherein after whole blood sample DNA for 6 times
4the epstein barr virus dna of IU, from amplification curve, baseline is smooth, and exponential region is obvious, the variation coefficient <2% of the Ct value that same sample detects for 6 times, concentration variation coefficient <50%, reproducible.Illustrate that test kit provided by the invention effectively can collect the viral DNA in white corpuscle.
Claims (4)
1. DNA extraction kit in a human whole blood white corpuscle, described test kit comprises erythrocyte cracked liquid and DNA extraction liquid, described erythrocyte cracked liquid comprises following component: concentration is the sodium-chlor of 5mmol/L, concentration is the Triton X-100 of 1.0% (v/v), concentration is the glucose of 320mmol/L, concentration is the Tutofusin tris of 0.01mol/L, surplus is sterilized water, and uses sodium hydroxide or hydrochloric acid soln adjust ph to make the pH value of described erythrocyte cracked liquid be 8.0 ~ 8.4;
Described DNA extraction liquid comprises the Repone K of 0.6 ~ 1.0mol/L, the ethylenediamine tetraacetic acid (EDTA) of 0.4 ~ 0.8mol/L, the sodium lauryl sulphate of 0.8 ~ 1.2% (m/v), surplus is sterilized water, and uses sodium hydroxide or hydrochloric acid soln adjust ph to make the pH value of described DNA extraction liquid be 8.0 ~ 8.4; Or described DNA extraction liquid comprises Sha graceful 0.01 ~ 0.5mmol/L, Repone K 20 ~ 300mmol/L of ancient India, sodium laurylsulfonate 0.01 ~ 2% and ethanol 0.05 ~ 1%.
2. test kit according to claim 1, is characterized in that, the pH value of described erythrocyte cracked liquid is 8.2.
3. test kit according to claim 1 and 2, it is characterized in that, the Repone K of 0.6 ~ 1.0mol/L is comprised at described DNA extraction liquid, the ethylenediamine tetraacetic acid (EDTA) of 0.4 ~ 0.8mol/L, the sodium lauryl sulphate of 0.8 ~ 1.2% (m/v), when surplus is sterilized water, sodium hydroxide or hydrochloric acid soln adjust ph is used to make the pH value of described DNA extraction liquid be 8.2.
4. the application of the test kit in claims 1 to 3 described in any one in fluorescent PCR detection technique.
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| CN107988210A (en) * | 2018-01-09 | 2018-05-04 | 厦门基源医疗科技有限公司 | A kind of method of rapid extraction blood and buccal swab genomic DNA |
| CN110804589A (en) * | 2018-08-06 | 2020-02-18 | 上海医药临床研究中心 | Separation and purification method of leucocyte |
| CN108977436A (en) * | 2018-08-10 | 2018-12-11 | 邵忠民 | Biotinylated nucleic acid DNA quick release extracts reagent and preparation method and application |
| CN109932359A (en) * | 2019-02-11 | 2019-06-25 | 张丽英 | A method of utilizing clostridium botulinum in ATP bioluminescence reaction detection food |
| CN109706143A (en) * | 2019-02-28 | 2019-05-03 | 武汉华大医学检验所有限公司 | A kind of extracting method of peripheral blood high molecular weight genomic DNA |
| CN110747255B (en) * | 2019-11-27 | 2023-08-22 | 郑州安图生物工程股份有限公司 | Pretreatment reagent and method for whole blood sample for nucleic acid detection |
| CN111808843A (en) * | 2020-07-21 | 2020-10-23 | 武汉海吉力生物科技有限公司 | DNA extraction kit and extraction method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101041846A (en) * | 2007-01-29 | 2007-09-26 | 中国科学院等离子体物理研究所 | Preparation method and usage for novel lipopeptide type biosurfactant surfactin |
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| CN102747036A (en) * | 2011-04-19 | 2012-10-24 | 北京同盛颐和生物科技有限公司 | Kit for in-vitro separation purification of karyocytes and removal of erythrocytes and use method of the kit |
-
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101041846A (en) * | 2007-01-29 | 2007-09-26 | 中国科学院等离子体物理研究所 | Preparation method and usage for novel lipopeptide type biosurfactant surfactin |
Non-Patent Citations (5)
| Title |
|---|
| 三种提取血细胞基因组DNA方法的比较;陈俊杰等;《遗传与疾病》;19881231;第5卷(第4期);第230页左栏第1节 * |
| 王小谦.全血基因组DNA 的提取和纯化方法比较.《河南科技大学学报（医学版）》.2005,第23卷(第1期),第13页1.2.1节. * |
| 闾宏伟等.四种微量全血DNA 提取方法的比较.《中国医学工程》.2004,第12卷(第5期),第44页第1.2节. * |
| 陈子祥等.一种快速高效提取全血基因组DNA 的实验方法.《中国现代医学杂志》.2010,第20卷(第22期),第3404-3405页第1.2.2-1.3.2节. * |
| 韩芬霞等.全血DNA 提取方法的改进及PCR 检测.《江苏农业科学》.2012,第40卷(第8期),第56页1.1-1.2节. * |
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