CN106404734A - Method of using silver nanocluster to detect potassium ion concentration in vitreous fluid to deduce death time - Google Patents
Method of using silver nanocluster to detect potassium ion concentration in vitreous fluid to deduce death time Download PDFInfo
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Abstract
The invention belongs to the technical field of medicine, and particularly relates to a method of using silver nanocluster to detect potassium ion concentration in vitreous fluid to deduce death time. The method includes that a novel DNA strand containing potassium ion aptamer is used as a template to synthesize DNA-AgNCs having fluorescent property, the system specifically recognizes K+ to quench fluorescence of the silver nanocluster, and a standard curve is drawn according to changing of the fluorescence; vitreous fluid to be detected is added into a DNA-AgNCs solution, potassium ions in the vitreous fluid lead to changing of the fluorescence of the silver nanocluster, the potassium ion concentration in the detected vitreous fluid is acquired by comparing with the standard curve; a standard curve between K+ concentration in a vitreous fluid sample and known death time is drawn, and a result has high uniformity to reported death time and vitreous fluid potassium ion relation. The method can be used for deducing death time by legal medical experts, has high specificity and sensitivity, is simple and quick to operate and low in cost and can be applied to detecting potassium ion content in other body fluids and water environment.
Description
Technical field
The invention belongs to prudence technical field is and in particular to a kind of silver nanoclusters detect potassium concentration in vitreous humor
And then the method for infering diing time.
Background technology
Potassium ion (K+) it is one of people's in-vivo content cation the abundantest, there is important physiological function to organism.
For keeping normal acid-base balance and the osmotic pressure of body, participate in various protein and carbohydrate metabolism institute is required；It is to maintain cell newly old
One of metabolism, important electrolyte of regulation body fluid pervious pressure, maintenance acid-base balance and holding cellular stress function.Its content is machine
The very important index of body physiological activity, simultaneously blood, the contents level of potassium ion can be used for Diagnosing Cardiac, kidney etc. in urine
The disease of aspect and the prognosis judging disease.
Additionally, in forensic identification, because malpractice etc. happens occasionally；Especially electrolyte disturbance, myocardial cell function
There is the relatively straightforward relation of ratio with the concentration of some cationes inside and outside cell membrane, wherein with the impact of calcium ion and potassium concentration
Substantially.Wherein especially with K in vitreous humor+Content is prudence study hotspot；In human body after death, K in vitreous humor+Content be in
Regular rising, extremely close with death time relation, by measuring K in vitreous humor+Content to infer that human body is dead when
Between；Have proven to K in vitreous humor simultaneously+Content is subject to outside contamination, the putrefaction of dead body and such environmental effects very little.Therefore, accurate
Really measurement K+Content has extremely important meaning to deduction Postmortem Interval.
K is measured in prior art+The method of content mainly has：Chemical assay, isotopic mass spectrometry, flame spectrometry,
Atom spectrophotography, enzyme process and ion selective electrode method etc..Wherein common detection methods are ion selective electrode method and fire
Flame photometry.
(1) flame spectrometry：The spectral intensity that the method is launched when being returned and be down to ground state using excited atom in flame is entered
Row content analysis, are a kind of emission spectrometry methods, can detect the K in vitreous humor, urine, serum, cerebrospinal fluid+Content.Often
The quantitative approach seen includes internal standard method and external standard method, and wherein external standard method general operation error is larger, and is of little use；Internal standard method will be marked
Add the internal standard of same concentrations to be measured in quasi- liquid and specimen, be usually added into lithium internal standard, subsequently measure lithium/potassium current ratio
Value, rather than the electric current of independent measurement potassium ion, so just can reduce error, improve the accuracy of measurement.Itself the disadvantage is that, fire
In detection process, sensitivity is affected flame photometry by gaseous-pressure, the pressure of combustion-supporting gas, specimen sample introduction speed, and specimen adds
To fully mix after distilled water, the sample otherwise analyzed and actual value have certain deviation.Additionally, the maximum deficiency of flame spectrometry exists
In combustion gas such as used propane, bring potential safety hazard to laboratory.
(2) chemical assay：Main using ionophore, ring crown compound is measured again, has size due to inside ring
The cavity structure differing, its internal oxygen atom electronics is to can be with metal ion-chelant.Different size of cavity is alternative to be combined
The metal ion of different-diameter, thus measure ion concentration.But anion, solvent, crown compound concentration, thickness of liquid film and
The migration rate impact in compound is larger on metal ion for the factors such as temperature, is easily caused measurement result inaccurate.
(3) ion selective electrode method：Urine, blood, K+ determination in water are carried out on instrumentation.Its principle is：
Ion-selective electrode is a class chemical sensor, within the specific limits, fixed ionic activity given in its current potential and solution
Logarithm is in linear relation, can be utilized this linear relationship to measure liquor sample, the color of solution and turbidity typically will not be to measurements
Result has an impact.The method is divided into direct mensure and Indirect Determination, at present frequently with the latter's mensure again.Due to Indirect Determination
Measure after determinand is diluted, gained ionic activity is closer to ion concentration.The main electrode adopting has：Solid phase electrode, liquid
Membrane electrode, glass-membrane electrode etc..But these electrodes can be aging with use time, expensive.
Content of the invention
The problem existing for prior art, the present invention provides potassium concentration in a kind of silver nanoclusters detection vitreous humor
The method of infering diing time it is therefore an objective to potassium concentration is measured by the DNA-AgNCs fluorescent probe being synthesized based on DNA,
Make K+ determination easier, more rapid, more accurate.
The present invention provide silver nanoclusters detect vitreous humor in potassium concentration infering diing time method, according to
Lower step is carried out：
(1) prepare fluorescent DNA-AgNCs：Design containing potassium ion aptamers first and the DNA of AgNCs can be loaded,
AgNO is added in DNA3Solution, adds NaHB after vibration4Solution, puts into refrigerator 3 hours after vibrating again, obtains brown color liquid,
I.e. fluorescent DNA-AgNCs；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared：Prepare variable concentrations potassium salt soln, be added into step
Suddenly, in the fluorescent DNA-AgNCs solution of (1), reaction measures its fluorescence intensity at 630nm after a period of time, obtains potassium ion
Concentration and the standard curve of fluorescence intensity；
(3) actual glass body fluid sample potassium concentration detection：By the vitreous humor to be measured of known death time add to
In the fluorescent DNA-AgNCs solution of step (1), reaction measures its fluorescence intensity at 630nm, according to preparation after a period of time
The potassium concentration of step (2) and Standardization curve for fluorescence intensity draw K in institute's test sample basis+Concentration；
(4) potassium concentration and death time standard curve are prepared：According to the known death time measured in step (3)
Vitreous humor K+Concentration and the death time of known sample, draw linear relationship chart, prepared potassium concentration and death time
Standard curve；
(5) corrupt corpse：Measure the vitreous humor K to be measured of unknown death time+Concentration, according to the potassium of step (4)
Ion concentration and death time standard curve, thus be inferred to this death time of institute's test sample.
Wherein, described containing potassium ion aptamers and can to load the DNA of AgNCs be that can to form G- tetrameric
DNA, selected from nucleotide sequence table id NO.1, nucleotide sequence table id NO.2, nucleotide sequence table id NO.3, nucleotide
Sequence table id NO.4, nucleotide sequence table id NO.5, nucleotide sequence table id NO.6, nucleotide sequence table id NO.7, core
Nucleotide sequence table id NO.8, nucleotide sequence table id NO.9, nucleotide sequence table id NO.10, nucleotide sequence table id
NO.11, nucleotide sequence table id NO.12, DNA sequence shown in nucleotide sequence table id NO.13..
Described DNA and AgNO3、NaHB4Molar concentration rate is 1:2～10:2～10, preferably 1:6:6.
The pH value of described fluorescence AgNCs solution is 6-7.5.
Described potassium salt includes potassium chloride, potassium nitrate and potassium bromide and other potassium-containing compound.
Described solution to be measured is vitreous humor, blood, urine or other liquid of human or animal.
The response time of described step (2) and step (3) is 0.1～4 hour.
In described utilization silver nanoclusters detection liquid, potassium concentration is used for the accurate supposition death time in forensic identification.
Methods described is used for detecting other compositions in vitreous humor, including ATP, hypoxanthine, magnesium ion.
Compared with prior art, the feature of the present invention and beneficial effect are：
The silver nanoclusters of the present invention detect the ultimate principle of the method for potassium concentration infering diing time in vitreous humor
It is that potassium concentration is measured based on DNA synthetic DNA-AgNCs fluorescent probe, formed with Ag using the N atom in DNA monomer
Stronger coordinate bond, sends hyperfluorescence after amino specificity connects DNA-AgNCs, is by AgNO in the present invention3、NaHB4Press
Certain proportion is coupled on target dna by chemical bond, obtains fluorescent DNA-AgNCs after Physical temperature-lowering, subsequently this fluorescence nano
Race specific chelating K again+Make its fluorescent quenching, the change according to fluorescence can measure K+Concentration.Potassium ion solution to be measured is dripped
In above-mentioned fluorescent DNA-AgNCs solution, detect its change in fluorescence, by compared with obtained standard curve, draw by
Survey the Potassium Level of body fluid.
It is an advantage of the invention that：
(1) present invention using DNA specific recognition and combines K+, form the G- tetramer conformation of regulation and control, can not be subject to it
The impact of his ion, has high specificity to potassium ion.
(2) present invention is used DNA-AgNCs as fluorescent probe, to K+The G- tetramer conformation of regulation and control is very sensitive, tool
There is higher sensitivity.
(3) the DNA-AgNCs fluorescent probe based on DNA of present invention design, only needs fluorescence spectrophotometer, cost is relatively low,
Can be widely popularized.
(4) present invention for K+The condition that the DNA-AgNCs fluorescent probe preparation process based on DNA of detection requires
Easy to control, operation is succinct, and low energy consumption, is process for cleanly preparing, can be produced in batches.
(5) present invention for K+The DNA-AgNCs fluorescent probe based on DNA of detection, agents useful for same is commonly
Chemical reagent safety non-toxic, environment friendly and pollution-free, can extensively apply.
(6) agents useful for same species of the present invention is less, simple operationss, quick and with low cost；Material stability is good, when can be long
Between storage it is ensured that measure effect.
(7) K+ determination method provided by the present invention not only can measure human body, blood, urine, glass in animal body
K in the body fluid such as glass body fluid+Concentration, can also be measured to the potassium concentration in other samples such as water quality, is widely used.
(8) provided by the present invention measure vitreous humor in potassium concentration method, can quickly and accurately measure potassium from
Fluorescent technique is applied to forensic identification by sub- concentration first, is that legal medical expert worker infers the dead death time provider exactly
Method.
Brief description
Fig. 1 is that the present invention prepares measurement fluorescence intensity schematic diagram at 630nm containing potassium solution during standard curve；
Fig. 2 is the canonical plotting being obtained according to Fig. 1；
Fig. 3 is the canonical plotting in the embodiment of the present invention 1；
Fig. 4 is the canonical plotting in the embodiment of the present invention 2；
Fig. 5 is the canonical plotting in the embodiment of the present invention 3.
Specific embodiment
Below by way of being embodied as case, the present invention will be described, but protection scope of the present invention is not limited thereto.
Embodiment 1：
The silver nanoclusters of the present embodiment detect the method for potassium concentration infering diing time in vitreous humor according to following
Step is carried out：
(1) prepare fluorescent DNA-AgNCs：Take the DNA solution of 50 μM of 90 μ L with 12000rpm under the conditions of 4 DEG C, take 50 μ L
Bottom liquid, to centrifuge tube, adds the AgNO of 500 μM of 54 μ L in centrifuge tube3Solution and 102 μ L secondary waters, shake at room temperature
Swing 30 minutes；Subsequently, 500 μM of NaHB being prepared with frozen water of 54 μ L are added wherein4Solution, concussion put into 4 DEG C of ice after 5 minutes
Case stores 3 hours, obtains brown color liquid, i.e. fluorescent DNA-AgNCs solution, and the pH of solution is 7.4, is sub-packed in centrifuge tube, often
Part Deca 30 μ LDNA-AgNCs solution, standby；
DNA nucleotides sequence used in this example is classified as DNA sequence shown in nucleotide sequence table id NO.6；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared：Compound concentration be 0.1,0.4,0.8,1.2,1.8,
3.0th, 5.0,7.5,10.0, the potassium nitrate solution of 20.0mM, is added into the fluorescent DNA-AgNCs solution of step (1), treats
After solution reaction one minute, measure its fluorescence intensity at 630nm, draw solution K+Concentration and the linear pass of fluorescence intensity
System, prepared standard curve；
(3) actual glass body fluid sample potassium concentration detection：By vitreous humor to be measured for 20 μ L (known dead 13.2 hours
Obtaining afterwards) sample solution adds to the fluorescent DNA-AgNCs solution of step (1), and add 100 μ L secondary water dilutions, 1 minute
Measure its fluorescence intensity at 630nm afterwards.
(4) potassium concentration and death time standard curve are prepared：According to the known death time measured in step (3)
Vitreous humor K+Concentration and the death time of known sample, measure a series of vitreous humor K of difference death times+Concentration, draws
The standard curve of linear relationship chart, prepared potassium concentration and death time；
(5) corrupt corpse：K in drawing institute's test sample originally according to the standard curve of preparation process (2)+Concentration is 8.6mM；
The potassium concentration of step (4) and death time standard curve, thus infer that the death time obtaining detected sample is 13.6
Hour, substantially it is consistent with the actual sample death time situation of the present embodiment.Take this vitreous humor sample solution standard again
EDTA titrimetry is measured, and obtains K+Concentration is 8.71mM；Measuring result error is less than 2%.Measurement result has good
Concordance, illustrate the inventive method can quantitative measurement variable concentrations K+, concentration ranges are 0.1～20.0mM..
Embodiment 2：
The silver nanoclusters of the present embodiment detect the method for potassium concentration infering diing time in vitreous humor according to following
Step is carried out：
(1) prepare fluorescent DNA-AgNCs：Take the DNA solution of 50 μM of 100 μ L with 12000rpm under the conditions of 4 DEG C, take 60 μ
L bottom liquid, to centrifuge tube, adds the AgNO of 500 μM of 108 μ L in centrifuge tube3Solution and 204 μ L secondary waters, at room temperature
Concussion 30 minutes；Subsequently, 500 μM of NaHB being prepared with frozen water of 108 μ L are added wherein4Solution, concussion put into 4 DEG C after 5 minutes
Refrigerator storage 3 hours, obtains brown color liquid, i.e. fluorescent DNA-AgNCs solution, and the pH of solution is 7.4, is sub-packed in centrifuge tube,
Every part of Deca 50 μ L DNA-AgNCs solution, standby；
DNA nucleotides sequence used in this example is classified as DNA sequence shown in nucleotide sequence table id NO.6；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared：Compound concentration be 0.1,0.8,2.4,5.0,10.0,
18.0th, 25.0, the 30.0, potassium nitrate solution of 40.0mM, by 20 μ L potassium nitrate solutions add to the fluorescent DNA of step (1)-
In AgNCs solution and add 100 μ L secondary waters dilution, after solution reaction one minute, measure its fluorescence at 630nm strong
Degree, draws solution K+Concentration and the linear relationship of fluorescence intensity, prepared standard curve；
(3) actual glass body fluid sample potassium concentration detection：By vitreous humor to be measured for 20 μ L (known dead 19.5 hours
Obtaining afterwards) sample adds to the fluorescent DNA-AgNCs solution of step (1), and add 100 μ L secondary water dilutions, survey after 1 minute
Measure its fluorescence intensity at 630nm.
(4) potassium concentration and death time standard curve are prepared：According to the known death time measured in step (3)
Vitreous humor K+Concentration and the death time of known sample, measure a series of vitreous humor K of difference death times+Concentration, draws
The standard curve of linear relationship chart, prepared potassium concentration and death time；
(5) corrupt corpse：K in drawing institute's test sample originally according to the standard curve of preparation process (2)+Concentration is
11.8mM；The potassium concentration of step (4) and death time standard curve, thus infer the death time obtaining detected sample
For 18.9 hours, it is consistent with the actual sample death time situation of the present embodiment.Take this vitreous humor sample standard again
EDTA titrimetry is measured, and obtains K+Concentration is 11.67mM；Measuring result error is less than 1%.Measurement result has good
Concordance, illustrate the inventive method can quantitative measurement variable concentrations K+, concentration ranges are 0.1～40.0mM.
Embodiment 3：
The silver nanoclusters of the present embodiment detect the method for potassium concentration infering diing time in vitreous humor according to following
Step is carried out：
(1) prepare fluorescent DNA-AgNCs：Take the DNA solution of 50 μM of 100 μ L with 12000rpm under the conditions of 4 DEG C, take 60 μ
L bottom liquid, to centrifuge tube, adds the AgNO of 500 μM of 108 μ L in centrifuge tube3Solution and 204 μ L secondary waters, at room temperature
Concussion 30 minutes；Subsequently, 500 μM of NaHB being prepared with frozen water of 108 μ L are added wherein4Solution, concussion put into 4 DEG C after 5 minutes
Refrigerator storage 3 hours, obtains brown color liquid, i.e. fluorescent DNA-AgNCs solution, and the pH of solution is 7.4, is sub-packed in centrifuge tube,
Every part of Deca 50 μ L DNA-AgNCs solution, standby；
DNA nucleotides sequence used in this example is classified as DNA sequence shown in nucleotide sequence table id NO.6；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared：Compound concentration be 0.1,0.8,2.4,5.0,10.0,
18.0th, 25.0, the 30.0, potassium nitrate solution of 40.0mM, by 20 μ L potassium nitrate solutions add to the fluorescent DNA of step (1)-
In AgNCs solution and add 100 μ L secondary waters dilution, after solution reaction one minute, measure its fluorescence at 630nm strong
Degree, draws solution K+Concentration and the linear relationship of fluorescence intensity, prepared standard curve；
(3) actual glass body fluid sample potassium concentration detection：By vitreous humor to be measured for 20 μ L (after known dead 6 hours
Obtaining) sample adds to the fluorescent DNA-AgNCs solution of step (1), and add 100 μ L secondary water dilutions, measure after 1 minute
Its fluorescence intensity at 630nm.
(4) potassium concentration and death time standard curve are prepared：According to the known death time measured in step (3)
Vitreous humor K+Concentration and the death time of known sample, measure a series of vitreous humor K of difference death times+Concentration, draws
The standard curve of linear relationship chart, prepared potassium concentration and death time；
(5) corrupt corpse：K in drawing institute's test sample originally according to the standard curve of preparation process (2)+Concentration is 4.2mM；
The potassium concentration of step (4) and death time standard curve, thus infer that the death time obtaining detected sample is little for 6.3
When, it is consistent with the actual sample death time situation of the present embodiment.Take the EDTA titrimetry of this vitreous humor sample standard again
It is measured, obtain K+Concentration is 4.1mM；Measuring result error is less than 1%.Measurement result has good concordance, explanation
The inventive method can quantitative measurement variable concentrations K+, concentration ranges are 0.1～40.0mM.
Embodiment 4：
The detection method of the present embodiment is same as Example 1, only changes the DNA in embodiment into nucleotides sequence and is classified as core
DNA sequence shown in nucleotide sequence table id NO.1, final measurement equally can be realized to K+The quantitative determination of concentration, measures
A series of vitreous humor K of difference death times+Concentration, draws linear relationship chart, prepared potassium concentration and death time
Standard curve, measures the vitreous humor K to be measured of unknown death time+Concentration, bent with death time standard according to potassium concentration
Line, thus be inferred to this death time of institute's test sample.
Embodiment 5：
The detection method of the present embodiment is same as Example 2, only changes the DNA in embodiment into nucleotides sequence and is classified as core
DNA sequence shown in nucleotide sequence table id NO.2, final measurement equally can be realized to K+The quantitative determination of concentration, measures
A series of vitreous humor K of difference death times+Concentration, draws linear relationship chart, prepared potassium concentration and death time
Standard curve, measures the vitreous humor K to be measured of unknown death time+Concentration, bent with death time standard according to potassium concentration
Line, thus be inferred to this death time of institute's test sample.
Embodiment 6：
The detection method of the present embodiment is same as Example 3, only changes the DNA in embodiment into nucleotides sequence and is classified as core
The DNA of DNA sequence shown in nucleotide sequence table id NO.8, final measurement equally can be realized to K+The quantitation of concentration is surveyed
Fixed, measure a series of vitreous humor K of difference death times+Concentration, draws linear relationship chart, prepared potassium concentration and death
The standard curve of time, measures the vitreous humor K to be measured of unknown death time+Concentration, according to potassium concentration and death time
Standard curve, thus be inferred to this death time of institute's test sample.
Embodiment 7：
The detection method of the present embodiment is same as Example 1, only changes the DNA in embodiment into nucleotides sequence and is classified as core
The DNA of DNA sequence shown in nucleotide sequence table id NO.9, final measurement equally can be realized to K+The quantitation of concentration is surveyed
Fixed, measure a series of vitreous humor K of difference death times+Concentration, draws linear relationship chart, prepared potassium concentration and death
The standard curve of time, measures the vitreous humor K to be measured of unknown death time+Concentration, according to potassium concentration and death time
Standard curve, thus be inferred to this death time of institute's test sample.
Embodiment 8：
The detection method of the present embodiment is same as Example 2, only changes the DNA in embodiment into nucleotides sequence and is classified as core
DNA sequence shown in nucleotide sequence table id NO.5, final measurement equally can be realized to K+The quantitative determination of concentration, surveys
A series of vitreous humor K of fixed difference death time+Concentration, draws linear relationship chart, prepared potassium concentration and death time
Standard curve, measure the unknown death time vitreous humor K to be measured+Concentration, according to potassium concentration and death time standard
Curve, thus be inferred to this death time of institute's test sample.
Embodiment 9：
The detection method of the present embodiment is same as Example 3, only changes the DNA in embodiment into nucleotides sequence and is classified as core
DNA sequence shown in nucleotide sequence table id NO.13, final measurement equally can be realized to K+The quantitative determination of concentration, survey
A series of vitreous humor K of fixed difference death time+Concentration, draws linear relationship chart, prepared potassium concentration and death time
Standard curve, measure the unknown death time vitreous humor K to be measured+Concentration, according to potassium concentration and death time standard
Curve, thus be inferred to this death time of institute's test sample.
SEQUENCE LISTING
<110>Central South University
Gorgeous monarch, fourth
River, insults
Prunus mume var.bungo Makino, Lee
Continue peak, Cai
Yadong, Guo
Baiyi draws, Zha Laga
<120>A kind of method that silver nanoclusters detect potassium concentration infering diing time in vitreous humor
<130> 1.01
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<170> PatentIn version 3.5
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ccaaccacac caaccccccc ccccc 25
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Claims (9)
1. a kind of silver nanoclusters detect potassium concentration infering diing time in vitreous humor method it is characterised in that according to
Following steps are carried out：
(1) prepare fluorescent DNA-AgNCs：Design containing potassium ion aptamers first and the DNA of AgNCs can be loaded, in DNA
Middle addition AgNO3Solution, adds NaHB after vibration4Solution, puts into refrigerator 3 hours after vibrating again, obtains brown color liquid, that is, glimmering
Light DNA-AgNCs；
(2) potassium concentration and Standardization curve for fluorescence intensity are prepared：Prepare variable concentrations potassium salt soln, be added into step
(1), in fluorescent DNA-AgNCs solution, reaction measures its fluorescence intensity at 630nm after a period of time, obtains potassium ion dense
Degree and the standard curve of fluorescence intensity；
(3) actual glass body fluid sample potassium concentration detection：The vitreous humor to be measured of known death time is added to step
(1), in fluorescent DNA-AgNCs solution, reaction measures its fluorescence intensity at 630nm, according to preparation process after a period of time
(2) potassium concentration and Standardization curve for fluorescence intensity draw K in institute's test sample basis+Concentration；
(4) potassium concentration and death time standard curve are prepared：Glass according to the known death time measured in step (3)
Body fluid K+Concentration and the death time of known sample, draw linear relationship chart, the standard of prepared potassium concentration and death time
Curve；
(5) corrupt corpse：Measure the vitreous humor K to be measured of unknown death time+Concentration, dense according to the potassium ion of step (4)
Degree and death time standard curve, thus be inferred to this death time of institute's test sample.
2. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method it is characterised in that described containing potassium ion aptamers and can to load the DNA of AgNCs be that can to form G- tetrameric
DNA, selected from nucleotide sequence table id NO.1, nucleotide sequence table id NO.2, nucleotide sequence table id NO.3, nucleotide
Sequence table id NO.4, nucleotide sequence table id NO.5, nucleotide sequence table id NO.6, nucleotide sequence table id NO.7, core
Nucleotide sequence table id NO.8, nucleotide sequence table id NO.9, nucleotide sequence table id NO.10, nucleotide sequence table id
NO.11, nucleotide sequence table id NO.12, DNA sequence shown in nucleotide sequence table id NO.13.
3. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that described DNA and AgNO3、NaHB4Molar concentration rate is 1:2～10:2～10, preferably 1:6:6.
4. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that the pH value of described fluorescent DNA-AgNCs solution is 6-7.5.
5. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that described potassium salt includes potassium chloride, potassium nitrate, potassium bromide and other potassium-containing compound.
6. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that described solution to be measured is vitreous humor, blood, urine or other liquid of human or animal.
7. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that the response time of described step (2) and step (3) is 0.1～4 hour.
8. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that described utilization silver nanoclusters detect that in liquid, potassium concentration is used for accurate supposition death in legal medical expert's detection
Time.
9. a kind of silver nanoclusters according to claim 1 detect the side of potassium concentration infering diing time in vitreous humor
Method is it is characterised in that methods described is used for detecting other compositions in vitreous humor, including ATP, hypoxanthine, magnesium ion.
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| CN107290516A (en) * | 2017-05-31 | 2017-10-24 | 中南大学 | Silver nanoclusters probe based on sandwich structure is to A β1‑40The method of oligomer specificity fluorescents detection |
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| CN102866185A (en) * | 2012-09-06 | 2013-01-09 | 北京航空航天大学 | Method for detecting potassium ion by preparing biosensor based on G-quadruplex and gold nanoparticle |
| CN103913443A (en) * | 2014-04-23 | 2014-07-09 | 安徽师范大学 | Aptamer sensor based on DNA-Ag NCs (deoxyribonucleic acid-silver nanoclusters) as well as preparation method, application and detection method thereof |
| CN106018366A (en) * | 2016-05-09 | 2016-10-12 | 福建中医药大学 | Fluorescent DNA-silver nanocluster, and preparation method and application thereof |
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| CN102866185A (en) * | 2012-09-06 | 2013-01-09 | 北京航空航天大学 | Method for detecting potassium ion by preparing biosensor based on G-quadruplex and gold nanoparticle |
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| CN107290516A (en) * | 2017-05-31 | 2017-10-24 | 中南大学 | Silver nanoclusters probe based on sandwich structure is to A β1‑40The method of oligomer specificity fluorescents detection |
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