CN106086187A - The method of DTT interference when hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion - Google Patents
The method of DTT interference when hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion Download PDFInfo
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Abstract
The method of DTT interference when hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion, by containing mismatch site and have the recognition sequence causing the primer of nucleic acid constant-temperature amplification and template and mix by concentration 100nM equal-volume, recognition sequence combines to be measured containing can be formed after mercury ion in mercury solution can be by the stable hybrid structure of polymerase identification；To stablize hybrid structure and hydrogenperoxide steam generator, amplification substrate, archaeal dna polymerase KF, SYBR Green I and amplified reaction buffer hybrid reaction, under polymerase effect, cause amplified reaction, produce double-stranded DNA；SYBR Green I can be specific binding on double chain DNA molecule, is simultaneously emitted by fluorescence signal；Utilize solubility curve and agarose gel electrophoresis that sequence after amplification is characterized, confirming that hydrogen peroxide eliminates DTT interference, the present invention eliminates the interference that trace amount mercury ion is detected by DTT, has good system compatible, the most easy and simple to handle, do not rely on complex device.
Description
Technical field
The invention belongs to the trace detection of biological technical field mercury ion, eliminate nucleic acid constant-temperature particularly to hydrogen peroxide and expand
The method of DTT interference when increasing detection mercury ion.
Technical background
Heavy metal ion has at a relatively high stability in the environment, and is difficult to be degraded by microorganisms.They once enter
Environment, is difficult to naturally be repaired, and can constantly be enriched with in ecosystem and transmit, also can produce very even if trace is taken in
Big toxicity, is the significant threat of ecological balance and human health.The Typical Representative polluted as heavy metal ion, mercury ion is being given birth to
In fabric texture, accumulation causes DNA damage, affects the interaction of ligand-receptor, destroys immune system, causes a series of disease
Disease, such as brain injury, renal failure, various cognitions and dyskinesias etc..Therefore, in environment, the detection by quantitative of mercury ion has important
Researching value and realistic meaning.
The method of traditional detection mercury ion detecting have Atomic Absorption/emission spectrum, chromatography, inductive etc. from
Daughter mass spectrum, cold vapor atoms fluorescence spectrum, high performance liquid chromatography, anodic stripping voltammetry etc., but these methods depend on mostly
Relying in main equipment, experimental cost is high, needs professional to operate, and the preprocessing process of sample is loaded down with trivial details, and these all constrain
The extensive application in actually detected of these methods.
In recent years, the method carrying out detecting based on functionalization nucleic acid is widely developed.Nucleic acid is with its high stability, raw
The thing compatibility, easy labelling and modification, low cost, the advantages such as application is wide play more and more important in biological detection and biochemical analysis
Effect.Akira Ono is equal within 2004, finding and propose that mercury ion can be with the 3rd nitrogen-atoms on thymine alkali bases first
Proton substitution reaction is occurred to form stable T-Hg2+-T structure.This structure even can be with A-T alkali original in substituted nucleic acids sequence
Base pairing structure, makes the secondary structure of nucleic acid recombinate.The discovery of this special coordination structure improves to a great extent
The sensitivity of mercury ion detecting and specificity, make breakthrough contribution for heavy metal analysis.Have been developed in multiple based on this at present
The method of the detection of nucleic acids mercury ion of structure, the detection method such as including colorimetric, electrochemistry, electrochemiluminescence, fluorescence spectrum.But
Although colorimetry result naked eyes are visible but insufficient sensitivity is high, and the synthesis of nanometer gold is the most relatively complicated with set-up procedure.
But and the highly sensitive operation of electrochemical process is complicated, the time is longer, need to be equipped with price electrochemistry correlator costly
Device, meanwhile, although relevant fluorescence method is quicker and easy owing to not being amplified, but is relatively difficult to acquisition and makes us full
The sensitivity of meaning and detection range.
Nucleic acid constant-temperature amplification technology is development in recent years, it is possible to amplify rapidly a plurality of few core under constant temperature
Thuja acid strand.Compared to conventional amplification technology such as PCR, nucleic acid constant-temperature amplification technology not only has higher amplification efficiency, and
Need not heat circulating equipment, be increasingly used for DNA, microRNA, and other bioactive molecule and metal from
The detection of son.The method utilizing nucleic acid constant-temperature amplification technology for detection mercury ion is necessarily developed, compared to traditional detection method,
Mercury ion detecting can be realized quickly and efficiently.Application nucleic acid constant-temperature amplification technology for detection mercury ion is the most necessarily developed.
When application nucleic acid constant-temperature amplification technology carries out signal amplification, it is often necessary to use toolenzyme such as Klenow
Fragment polymerase KF.And in the storage liquid and optimal reaction buffer of existing polymerase, generally contain biological thiol divide
Son such as dithiothreitol, DTT DTT is to ensure rock-steady structure and the activity of enzyme.Research in the past shows, biological thiol molecule has with mercury ion
There is the strongest affinity interaction.The existence of biological thiol molecule can be competed with thymus pyrimidine, and trace amount mercury ion in capture system causes
Mercury ion reduces with thymus pyrimidine action effect, and the detection sensitivity causing trace amount mercury ion is on the low side, affects nucleic acid constant-temperature amplification
Method application in terms of mercury ion detecting.
Summary of the invention
For the defect overcoming above-mentioned prior art to exist, during present invention aims to nucleic acid constant-temperature amplification
The DTT impact on mercury ion detecting in polymerase KF, it is provided that when a kind of hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion
The method of DTT interference, utilizes hydrogen peroxide to eliminate dithiothreitol, DTT DTT, to promote that nucleic acid constant-temperature amplification technology is at hydrargyrum further
Extensive application in ion detection.
In order to achieve the above object, the present invention is achieved through the following technical solutions:
The method of DTT interference when hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion, comprises the following steps:
(1) by the recognition sequence of the primer that containing mismatch site and there is initiation nucleic acid constant-temperature amplification and template by concentration
100nM equal-volume mixes, and recognition sequence combines to be measured containing can be formed after mercury ion in mercury solution can be by polymerase identification
Stablize hybrid structure；
(2) by combine stable hybrid structure after mercury ion and hydrogenperoxide steam generator, amplification substrate, archaeal dna polymerase KF,
SYBR Green I and amplified reaction buffer hybrid reaction, reaction temperature is 37 DEG C, and the time is 15min；In mixture respectively
Individual concentration of component is respectively as follows: primer concentration 100nM, template concentrations 100nM, concentration of hydrogen peroxide 0.01%-0.5%, the amplification end
Thing 40 μMs, archaeal dna polymerase KF 0.5U, SYBR Green I concentration are 2 × SYBR Green I, and amplified reaction buffer is 10mM
Tris,75mM KAc,10mM Mg(Ac)2, pH=7.9；Primer and template expose 3 ' ends after forming stable hybrid structure,
Cause amplified reaction under polymerase effect, produce double-stranded DNA；SYBR Green I can be specific binding at double chain DNA molecule
On, it is simultaneously emitted by fluorescence signal；
(3) utilize solubility curve and agarose gel electrophoresis that sequence after amplification is characterized, it was demonstrated that hydrogen peroxide eliminates
DTT interference, the condition used by solubility curve that makes is: scan fluorescence signal in the range of 60-95 DEG C with real-time fluorescence PCR；Fine jade
Sepharose electrophoresis characterizes condition: 2% agarose gel concentration, under 60V voltage, and electrophoresis 30min.
The described primer that containing mismatch site and there is initiation nucleic acid constant-temperature amplification and the recognition sequence of template, its nucleotide
Sequence (5 ' to 3 '):
Mismatched primers: TAGAGGTT
Template: TGAGGCTAGAGCGAGCTGAGGCGGATATGGAATACTACCTCTAAA.
Described containing mercury solution: Mercury pernitrate..
Described amplification substrate is commercially available dNTPs mixture.
Compared with prior art, the present invention has a following useful technique effect:
1. relatively more traditional mercury ion detecting method have Atomic Absorption/emission spectrum, chromatography, inductive etc. from
Daughter mass spectrum, cold vapor atoms fluorescence spectrum, high performance liquid chromatography, anodic stripping voltammetry etc., but these methods depend on mostly
Relying in main equipment, experimental cost is high, needs professional to operate, and the preprocessing process of sample is loaded down with trivial details, and these all constrain
The extensive application in actually detected of these methods.
2. although nucleic acid constant-temperature amplification method is easy and simple to handle in the past, but owing to polymerase used generally containing DTT, should
Molecule and mercury ion have the strongest affinity, trace amount mercury ion can be disturbed to detect, cause sensitivity low, and poor reproducibility etc. is asked
Topic.The present invention is directed to the interference problem of DTT, the one of proposition in the application in terms of mercury ion detecting of the nucleic acid constant-temperature amplification technology
Utilize hydrogen peroxide to eliminate the simple and easy to do solution of DTT, can effectively eliminate the interference of DTT molecule in polymerase, improve
Nucleic acid amplification efficiency, has important application prospect.
Accompanying drawing explanation
Fig. 1 is the reaction process principle schematic of the present invention.
Fig. 2 is the hydrogen peroxide reinforced effects schematic diagram to existing nucleic acid amplification technologies.
Fig. 3 is that checking DTT is on mercury ion detecting impact and the schematic diagram of hydrogen peroxide elimination effect.
Detailed description of the invention
Below in conjunction with the accompanying drawings the present invention is done narration in detail.
Embodiment one
The method of DTT interference when hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion, comprises the following steps:
(1) by the recognition sequence of the primer that containing mismatch site and there is initiation nucleic acid constant-temperature amplification and template by concentration
100nM equal-volume mixes, and recognition sequence combines to be measured containing can be formed after mercury ion in mercury solution can be by polymerase identification
Stablize hybrid structure；
(2) by combine stable hybrid structure after mercury ion and hydrogenperoxide steam generator, amplification substrate, archaeal dna polymerase KF,
SYBR Green I and amplified reaction buffer hybrid reaction, reaction temperature is 37 DEG C, and the time is 15min；In mixture respectively
Individual concentration of component be respectively as follows: primer concentration 100nM, template concentrations 100nM, concentration of hydrogen peroxide 0.01%, amplification substrate 40 μMs,
Archaeal dna polymerase KF 0.5U, SYBR Green I concentration are 2 × SYBR Green I, and amplified reaction buffer is 10mM Tris,
75mM KAc,10mM Mg(Ac)2, pH=7.9；Primer and template expose 3 ' ends, at polymerase after forming stable hybrid structure
Cause amplified reaction under effect, produce double-stranded DNA.SYBR Green I can be specific binding on double chain DNA molecule, simultaneously
Send fluorescence signal；
(3) utilize solubility curve and agarose gel electrophoresis that sequence after amplification is characterized, it was demonstrated that hydrogen peroxide eliminates
DTT interference, the condition used by solubility curve that makes is: scan fluorescence signal in the range of 60-95 DEG C with real-time fluorescence PCR.Fine jade
Sepharose electrophoresis characterizes condition: 2% agarose gel concentration, under 60V voltage, and electrophoresis 30min.
The described primer that containing mismatch site and there is initiation nucleic acid constant-temperature amplification and the recognition sequence of template, its nucleotide
Sequence (5 ' to 3 '):
Mismatched primers: TAGAGGTT
Template: TGAGGCTAGAGCGAGCTGAGGCGGATATGGAATACTACCTCTAAA.
Described containing mercury solution: Mercury pernitrate..
Described amplification substrate is commercially available dNTPs mixture.
From with reference to Fig. 1 it can be seen that contain mismatch site and have and cause the primer of nucleic acid constant-temperature amplification and the knowledge of template
Other sequence recognition after combining mercury ion, exposing can be by 3 ' end of polymerase identification, and under the effect of polymerase, DNA knows
3 ' ends of other element are with from as template initial amplification.But, owing to polymerase KF containing biological thiol molecule DTT, this point
Son can cause primer and template hybridization instability with recognition sequence competition binding mercury ion, cause nucleic acid amplification to be obstructed.And when to
When reaction system adds proper amount of hydrogen peroxide, peroxidating Hydrogen Energy oxidation DTT so that it is can not be in conjunction with mercury ion, therefore, by mercury ion
The hybrid structure energy stable existence triggered and formed, and make nucleic acid chains extend by nucleic acid amplification, produce the effect that signal amplifies.
Characterizing method one is to utilize the specific binding double-strandednucleic acid of fluorescent dye SYBR Green I, scans dissolving with real-time fluorescence PCR
Curve；Two is to utilize agarose gel electrophoresis to characterize double-stranded products.
With reference to Fig. 2, compare the hydrogen peroxide solubility curve to existing nucleic acid amplification technologies reinforced effects.Curve a, b are respectively
Represent reaction system background when without hydrogen peroxide and signal.Curve c, d represent interpolation peroxidating in reaction system
Background during hydrogen and signal.Illustration represents the electrophoresis pattern of the most corresponding a, b, c, d.As seen from the figure, only when detection hydrargyrum
Time in the reaction system of ion containing hydrogen peroxide, obvious fluorescence signal could be produced.Compared to being not added with hydrogen peroxide
Reaction system, fluorescence signal strengthens nearly 6 times.Can also draw from electrophoresis pattern, only when peroxide in the system of detection mercury ion
In the presence of changing hydrogen, amplification could be passed through and produce double-stranded products.Thus proving, amplified reaction only triggers at mercury ion, and exists
Could efficiently carry out during hydrogen peroxide, show that reaction system exists interference simultaneously, affect the stable knot of mercury ion and recognition sequence
Close.
With reference to Fig. 3, checking DTT is on mercury ion detecting impact and the elimination effect of hydrogen peroxide.For confirmatory reaction system
Mercury ion detecting is disturbed from DTT.Impact and the hydrogen oxide of mercury ion detecting system fluorescence signal are eliminated by relatively DTT
The change of system fluorescence signal after DTT.Curve a, b represent that reaction system is being added after hydrogen peroxide respectively, background and signal
Fluorescence intensity.Curve c, d add after DTT in being illustrated respectively in the reaction system (a, b) adding hydrogen peroxide, background and signal
Fluorescence intensity.Curve e, f represent respectively, in the reaction system (c, d) of above-mentioned interpolation DTT, after again adding hydrogen peroxide, and the back of the body
Scape and the fluorescence intensity of signal.It can be seen that after reaction system adds hydrogen peroxide first, mercurous ionic system produces
Raw obvious fluorescence signal, fluorescence signal strengthens nearly 6 times, shows the hydrogen peroxide potentiation to amplified reaction.And when reaction
After being individually added into DTT in system, signal substantially reduces, and close with background, shows that DTT is really and recognition sequence competition binding hydrargyrum
Ion, causes mercury ion with recognition sequence stable bond, and then amplified reaction can not be caused not carry out.When to adding DTT's
When reaction system adds hydrogen peroxide again, it can be seen that fluorescence signal recovers again, signal strengthens nearly 4 times, shows strictly
Being had the DTT of reducing property by hydrogen peroxide oxidation, reduce the DTT interference to the detection of mercury ion, amplified reaction is able to efficiently
Carry out.
Embodiment two
In the present embodiment, remaining step is identical, and difference is, the concentration of hydrogen peroxide in mixture described in step (2) is
0.5%.
Testing result shows, compared to being not added with the reaction system of hydrogen peroxide, fluorescence signal strengthens nearly 5 times.
Embodiment three
In the present embodiment, remaining step is identical, and difference is, the concentration of hydrogen peroxide in mixture described in step (2) is
0.2%.
Testing result shows, compared to being not added with the reaction system of hydrogen peroxide, fluorescence signal strengthens nearly 6 times.
Comparative example one
In the present embodiment, remaining step is identical, and difference is, the concentration of hydrogen peroxide in mixture described in step (2) is
0.005%.
Testing result shows, compared to being not added with the reaction system of hydrogen peroxide, fluorescence signal strengthens nearly 4 times, but low
In the effect of 0.01%.
Comparative example two
In the present embodiment, remaining step is identical, and difference is, the concentration of hydrogen peroxide in mixture described in step (2) is
0.06%.
Testing result shows, compared to the reaction system of interpolation hydrogen peroxidase 10 .05%, fluorescence signal strengthens nearly 4 times, but
Be less than 0.05% effect a lot.
Comprehensive Correlation example, therefore the optium concentration that hydrogen peroxide adds is at 0.01%-0.5%.
Claims (5)
1. the method for DTT interference when hydrogen peroxide eliminates nucleic acid constant-temperature amplification detection mercury ion, it is characterised in that include following step
Rapid:
(1) by the recognition sequence of the primer that containing mismatch site and there is initiation nucleic acid constant-temperature amplification and template by concentration 100nM
Equal-volume mixes, and recognition sequence combines to be measured containing can be formed after mercury ion in mercury solution can be miscellaneous by stablizing of polymerase identification
Knot structure；
(2) the stable hybrid structure after mercury ion and hydrogenperoxide steam generator, amplification substrate, archaeal dna polymerase KF, SYBR will be combined
Green I and amplified reaction buffer hybrid reaction, reaction temperature is 37 DEG C, and the time is 15min；Each component in mixture
Concentration be respectively as follows: primer concentration 100nM, template concentrations 100nM, concentration of hydrogen peroxide 0.01%-0.5%, amplification substrate 40 μMs,
Archaeal dna polymerase KF 0.5U, SYBR Green I concentration are 2 × SYBR Green I, and amplified reaction buffer is 10mM Tris,
75mM KAc,10mM Mg(Ac)2, pH=7.9；Primer and template expose 3 ' ends, at polymerase after forming stable hybrid structure
Cause amplified reaction under effect, produce double-stranded DNA；SYBR Green I can be specific binding on double chain DNA molecule, simultaneously
Send fluorescence signal；
(3) utilize solubility curve and agarose gel electrophoresis that sequence after amplification is characterized, it was demonstrated that hydrogen peroxide eliminates DTT
Interference, the condition used by solubility curve that makes is: scan fluorescence signal in the range of 60-95 DEG C with real-time fluorescence PCR；Agarose coagulates
Gel electrophoresis characterizes condition: 2% agarose gel concentration, under 60V voltage, and electrophoresis 30min.
The method of DTT interference when hydrogen peroxide the most according to claim 1 eliminates nucleic acid constant-temperature amplification detection mercury ion, its
It is characterised by,
The described primer that containing mismatch site and there is initiation nucleic acid constant-temperature amplification and the recognition sequence of template, its nucleotide sequence
(5 ' to 3 '):
Mismatched primers: TAGAGGTT
Template: TGAGGCTAGAGCGAGCTGAGGCGGATATGGAATACTACCTCTAAA.
The method of DTT interference when hydrogen peroxide the most according to claim 1 eliminates nucleic acid constant-temperature amplification detection mercury ion, its
It is characterised by, described containing mercury solution: Mercury pernitrate..
The method of DTT interference when hydrogen peroxide the most according to claim 1 eliminates nucleic acid constant-temperature amplification detection mercury ion, its
Being characterised by, described amplification substrate is commercially available dNTPs mixture.
The method of DTT interference when hydrogen peroxide the most according to claim 1 eliminates nucleic acid constant-temperature amplification detection mercury ion, its
It is characterised by, described concentration of hydrogen peroxide 0.01%.
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| CN201610460402.9A CN106086187B (en) | 2016-06-22 | 2016-06-22 | Method for eliminating DTT interference during detection of mercury ions by nucleic acid isothermal amplification by using hydrogen peroxide |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102778492A (en) * | 2012-07-13 | 2012-11-14 | 首都师范大学 | Electrochemical transducer for mercury ion detection and manufacturing method and detection method thereof |
| CN103305612A (en) * | 2013-06-04 | 2013-09-18 | 西安交通大学 | Lead ion detection kit based on constant-temperature cascade nucleic acid amplification and detection method of lead ion detection kit |
| CN103760205A (en) * | 2014-02-26 | 2014-04-30 | 长沙理工大学 | Method for dithiothreitol membrane mercury ion selective electrode |
| CN104198557A (en) * | 2014-09-03 | 2014-12-10 | 大连理工大学 | Enzyme based electrochemical biosensing method applied to detection of trace mercury ions |
| CN105256033A (en) * | 2015-10-22 | 2016-01-20 | 西安交通大学 | Mercuric ion detection kit based on constant-temperature cascading nucleic acid amplification and detection method thereof |
-
2016
- 2016-06-22 CN CN201610460402.9A patent/CN106086187B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102778492A (en) * | 2012-07-13 | 2012-11-14 | 首都师范大学 | Electrochemical transducer for mercury ion detection and manufacturing method and detection method thereof |
| CN103305612A (en) * | 2013-06-04 | 2013-09-18 | 西安交通大学 | Lead ion detection kit based on constant-temperature cascade nucleic acid amplification and detection method of lead ion detection kit |
| CN103760205A (en) * | 2014-02-26 | 2014-04-30 | 长沙理工大学 | Method for dithiothreitol membrane mercury ion selective electrode |
| CN104198557A (en) * | 2014-09-03 | 2014-12-10 | 大连理工大学 | Enzyme based electrochemical biosensing method applied to detection of trace mercury ions |
| CN105256033A (en) * | 2015-10-22 | 2016-01-20 | 西安交通大学 | Mercuric ion detection kit based on constant-temperature cascading nucleic acid amplification and detection method thereof |
Non-Patent Citations (5)
| Title |
|---|
| HIDEHITO URATA等: "Incorporation of Thymine Nucleotides by DNA Polymerases through T–HgII–T Base Pairing", 《ANGEWANDTE CHEMIE》 * |
| TAO LI等: "G-quadruplex-based DNAzyme for sensitive mercury detection with the naked eye", 《THE ROYAL SOCIETY OF CHEMISTRY》 * |
| WANGREN等: "Label-free colorimetric detection of Hg2+ based on Hg2+-triggered exonuclease III-assisted target recycling and DNAzyme amplification", 《BIOSENSORS AND BIOELECTRONICS》 * |
| XIAO ZHU等: "Ultrasensitive and selective detection of mercury(II) in aqueous solution by polymerase assisted fluorescence amplification", 《BIOSENSORS AND BIOELECTRONICS》 * |
| YUE ZHAO等: "Fidelity quantification of mercury(II) ion via circumventing biothiols-induced sequestration in enzymatic amplification system", 《THE ROYAL SOCIETY OF CHEMISTRY》 * |
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