CN102643917A - Lead ion detection test paper and preparation method thereof - Google Patents
Lead ion detection test paper and preparation method thereof Download PDFInfo
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- CN102643917A CN102643917A CN2012101247360A CN201210124736A CN102643917A CN 102643917 A CN102643917 A CN 102643917A CN 2012101247360 A CN2012101247360 A CN 2012101247360A CN 201210124736 A CN201210124736 A CN 201210124736A CN 102643917 A CN102643917 A CN 102643917A
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Abstract
The invention discloses a lead ion detection test paper and a preparation method thereof. The test paper consists of a bar-type base plate, a sample pad, a glass fiber combination pad, a specifically enveloped nitrocellulose membrane and a water absorption pad, wherein the sample pad, the glass fiber combination pad, the specifically enveloped nitrocellulose membrane and the water absorption pad are lapped on the bar-type base plate in sequence; the glass fiber combination pad is coated with a special bar-type code DNA-nano gold-deoxyribozyme probe; the nitrocellulose membrane is provided with a quality control line C and a detection line T; and pairing can be formed between a basic group capturing DNA and bar-type code DNA basic group on the surface of the special bar-type code DNA-nano gold-deoxyribozyme probe. The preparation method comprises the following steps: the prepared sample pad, the glass fiber combination pad coated with the special bar-type code DNA-nano gold-deoxyribozyme probe, the specifically enveloped nitrocellulose membrane and the water absorption pad are lapped, are pasted on the base plate and are cut into a thin strip with the width of 3-5mm to obtain the lead ion detection test paper. The lead ion detection test paper has the advantage of capability of detecting the lead ion in the detection sample in high flexibility and high selectivity.
Description
Technical field
The present invention relates to heavy metal ion detection technique field, especially relate to a kind of lead ion test strip and preparation method thereof.
Background technology
Plumbous (Pb) is one of heavy metal, in vivo with environment in the residence time long, nerve system of human body is had heavy damage, to HUMAN HEALTH generation serious threat.The lead of occurring in nature is mainly with ionic species (Pb
2+) exist, historical facts or anecdotes existing to lead ion accurately, sensitive, fast on-the-spot detect extremely important.
The method that detects lead ion at present mainly contains: ultraviolet-visible spectrophotometry, atomic emission spectrometry, ICP-MS method, atomic absorption spectrometry, electrochemical process, ion chromatography, capillary electrophoresis, heavy metal rapid detector method, test strip method etc.Ultraviolet-visible spectrophotometry poor selectivity detects limit for height; Atomic emission spectrometry, ICP-MS method, atomic absorption spectrometry, electrochemical process, ion chromatography, capillary electrophoresis, accuracy and highly sensitive, but all need use specific instrument; Cost an arm and a leg; Complex operation, and require the testing staff to possess certain expertise, analysis cost is high; Can't be applied to on-the-spot the detection, be difficult to popularize; The heavy metal rapid detector method can be applicable to on-the-spot the detection, but sensitivity is not high, poor selectivity, and sample pretreatment is complicated.Compare with aforesaid method, the test strip method is quick, easy and other tool advantage in detecting at the scene with it, but based on the test strip of traditional chelating developer, poor selectivity, sensitivity is low.
Summary of the invention
Technical problem to be solved by this invention provides a kind of highly sensitive, selectivity good and cost is low lead ion test strip based on nanometer gold (AuNPs), DNAzyme (DNAzyme) and barcode DNA and preparation method thereof.
The present invention solves the problems of the technologies described above the technical scheme that is adopted: a kind of lead ion test strip; Form successively by the sample pad of overlap joint, the coating specificity barcode DNA-nanometer gold-glass fiber conjugate pad of DNAzyme probe, the nitrocellulose membrane and the absorbent pad of specified packet quilt by the bar shaped base plate with on the bar shaped base plate for described test strip; Described specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold; In the nanometer gold surface bonding some barcode DNA are arranged; On described nitrocellulose membrane; Encapsulate orthoscopic nature controlling line C line with strepto-affinity element; Encapsulate orthoscopic detection line T line with biotinylated captured dna and the plain bonded mixture of strepto-affinity, complementary pairing between the base of the barcode DNA of the base of described biotinylated captured dna and described specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of described specificity barcode DNA-nanometer gold-DNAzyme probe does
, the structure of described biotinylated captured dna is biotin-5'-TTTTTTTTTTTTTTT-3', the diameter of described nanometer gold is 13～30 nm.
Described sample pad is the glass fiber paper through 0.01～0.05 mol/L Tris-HCl damping fluid immersion treatment, and wherein to contain the mass percent of sucrose be 1～10% to the Tris-HCl damping fluid, and the pH value is 5.0～8.0.
The described specificity barcode DNA-nanometer gold-quantity for spray of DNAzyme probe solution on described glass fiber conjugate pad is 2.0～10.0 μ L/cm
2Described specificity barcode DNA-nanometer gold-DNAzyme probe solution be the 100 μ M sequences of 2 μ L be the 100 μ M sequences of DNA and the 2 μ L of biotin-3'-ACTGTAGAGAAGG rA TATCACAAAAAAAAAAAAAAA-5'-SH be the DNA of 5'-TGACATCTCTTCTCCGAGCCGGTCGAAATAGTG-3' mix the DNAzyme binding substances; 100 μ M sequences with 8 μ L are after HS-5'-AAAAAAAAAAAAAAA-3' barcode DNA and DNAzyme binding substances mix then; The 50 nM nano-Au solutions of 4 mL are added in the said mixture; Behind incubation 16 h; Centrifugal abandoning supernatant, deposition are dissolved in 50 μ L again and contain gained in the 25 mM Tris-HCl damping fluids of 100 mM NaCl and 8% sucrose.
It is 1.0～3.0 μ g/cm that described strepto-affinity element encapsulates consumption.
The plain bonded mixture of described biotinylated captured dna and strepto-affinity encapsulates orthoscopic detection line T line, and to prepare process following: the strepto-affinity cellulose solution of the biotinylated captured dna solution of the 100 μ M of 108 μ L and 2 mg/mL of 50 μ L mixes; Under the room temperature behind incubation 2 h; Centrifugal 10 min under 12000 rpm; Abandoning supernatant after dissolving again with the PBS damping fluid of 25 mM of 54 μ L, is sprayed at nitrocellulose membrane with 1～3 μ L/cm and forms detection line T line.
A kind of preparation method of lead ion test strip may further comprise the steps:
(1) preparation of sample pad
With glass fiber paper with 0.01～0.05 mol/L Tris-HCl damping fluid soak wetting after, promptly obtained sample pad down in dry 4～12 hours at 20 ℃～37 ℃, wherein to contain the mass percent of sucrose be 1～10% to the Tris-HCl damping fluid, the pH value is 5.0～8.0;
(2) preparation of the glass fiber conjugate pad of coating specificity barcode DNA-nanometer gold-DNAzyme probe
With glass fiber paper with 0.01～0.05 mol/L Tris-HCl damping fluid soak wetting after; After under 20 ℃～37 ℃ dry 4～12 hours; On every square centimeter glass fiber paper, evenly spray specificity barcode DNA-nanometer gold-DNAzyme probe solution of 2.0～10.0 μ L; After under 20 ℃～37 ℃ dry 2～5 hours; Promptly obtain applying the glass fiber conjugate pad of specificity barcode DNA-nanometer gold-DNAzyme probe, described specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme, and its other end is combined with nanometer gold; In the nanometer gold surface bonding some barcode DNA are arranged, the structure of described specificity barcode DNA-nanometer gold-DNAzyme probe does
Wherein to contain the mass percent of sucrose be 1～10% to the Tris-HCl damping fluid, and the pH value is 5.0～8.0, and the diameter of described nanometer gold is 13～30 nm;
(3) preparation of the nitrocellulose membrane of specified packet quilt
Using spray appearance appearance is that the strepto-affinity cellulose solution of 1 mg/mL evenly is sprayed on the nitrocellulose membrane with concentration; Drying is 2～8 hours under 25 ℃～37 ℃; Form orthoscopic nature controlling line C line; Evenly be sprayed at nitrocellulose membrane on the direction of the amount edge of 1～3 μ L/cm and nature controlling line C line parallel biotinylated captured dna and strepto-affinity element bonded complex solution with spray appearance appearance; Drying is 2～8 hours under 25 ℃～37 ℃, forms orthoscopic detection line T line, promptly obtains the nitrocellulose membrane of specified packet quilt; Can form complementary pairing between the base of the barcode DNA of the base of wherein said biotinylated captured dna and described specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of described barcode DNA is biotin-5'-TTTTTTTTTTTTTTT-3';
(4) sample pad that step (1) is obtained; The glass fiber conjugate pad of coating specificity barcode DNA-nanometer gold-DNAzyme probe that step (2) obtains; The nitrocellulose membrane of the specified packet quilt that step (3) obtains and absorbent pad overlap according to the order of sequence and are pasted on the base plate; Be cut into the wide slice of 3～5 mm, promptly get the lead ion test strip.
The preparation method of described specificity barcode DNA-nanometer gold-DNAzyme probe solution is following: the 100 μ M sequences of 2 μ L are that the 100 μ M sequences of DNA and the 2 μ L of biotin-3'-ACTGTAGAGAAGG rA TATCACAAAAAAAAAAAAAAA-5'-SH are that the DNA of 5'-TGACATCTCTTCTCCGAGCCGGTCGAAATAGTG-3' mixes; At 90 C water-baths, 5 min; Get the DNAzyme binding substances after naturally cooling to room temperature; Be after barcode DNA and the DNAzyme binding substances of HS-5'-AAAAAAAAAAAAAAA-3' mixes with the 100 μ M sequences of 8 μ L then, the 50 nM nano-Au solutions of 4 mL are added in the said mixture, behind incubation 16 h; Centrifugal 15 min of 12000 rpm; Abandoning supernatant, deposition is dissolved in 50 μ L again and contains in 100 mM NaCl and the 25 mM Tris-HCl damping fluids, after mixing; Promptly get specificity barcode DNA-nanometer gold-DNAzyme probe solution; The mass percent that wherein said Tris-HCl damping fluid contains sucrose is 1～10%, and the concentration that contains NaCl is 100 mM, and described Tris-HCl pH of buffer value is 5.0～8.0.
The plain bonded complex solution of described biotinylated captured dna and strepto-affinity process for preparation is following: the strepto-affinity element of the captured dna solution of the 100 μ M of 108 μ L and 2 mg/mL of 50 μ L mixes; Under the room temperature behind incubation 2 h; Centrifugal 10 min under 12000 rpm; Abandoning supernatant after dissolving again with the PBS damping fluid of 25 mM of 54 μ L, promptly obtains the plain bonded complex solution of biotinylated captured dna and strepto-affinity.
It is 1.0～3.0 μ L/cm that described strepto-affinity cellulose solution encapsulates consumption; The solvent of described strepto-affinity cellulose solution is concentration 0.01～0.05 a mol/L PBS damping fluid; Containing the NaCl mass percent in the described PBS damping fluid is 0.5～1%; Containing the sodium lauryl sulphate mass percent is 0.01～0.05%, and described PBS pH of buffer value is 5.0～8.0.
The lead ion test strip that the present invention proposes: apply Streptavidin on the nature controlling line; Apply the plain bonded mixture of biotinylated captured dna and strepto-affinity on the detection line; Apply an end on the pad and be combined with nanometer gold, the other end is combined with the specificity barcode DNA-nanometer gold-DNAzyme probe of vitamin H; One terminal modified vitamin H of specificity barcode DNA-nanometer gold-DNAzyme probe chain can with the Streptavidin that applies on the nature controlling line through vitamin H-Streptavidin effect generation specificity combination; So specificity barcode DNA-nanometer gold-DNAzyme probe is hunted down and rests on the nature controlling line; Its end of the chain bonded nanometer gold is assembled colour developing, reaches the Quality Control purpose of detection; When lead ion exists; DNAzyme double-spiral structure in specificity barcode DNA-nanometer gold-DNAzyme probe disintegrate (lead ion act in the DNAzyme structure rA place and from rA cut-out DNAzyme DNA chain); Nanometer gold is dropped from nature controlling line (C); The barcode DNA on nanometer gold surface and the captured dna on the detection line (T) take place to match stranded and are hunted down, and its end of the chain bonded nanometer gold is assembled colour developing, reaches the purpose that detects lead ion; Wherein the strepto-affinity on the detection line (T) is plain combines with biotinylated captured dna to form mixture through vitamin H-Streptavidin effect, reaches the purpose of the captured dna of fixed biologically elementization.
This test strip has reduced the consumption of DNAzyme, thereby only needs little amounts of Pb
2+Just can the DNAzyme double-spiral structure be disintegrated; Nanometer gold is dropped from nature controlling line (C); Improve detection sensitivity, introduce barcode DNA again first, a large amount of barcode DNA are attached to the nanometer gold surface; Nanometer gold is very easily captured by captured dna on detection line (T), further improve detection sensitivity.Sample liquid is detected with this test strip: if when the plumbum ion concentration in the sample liquid is higher than detectability, detection line and nature controlling line while exhibit red; If when not containing lead ion or plumbum ion concentration in the sample liquid and being lower than detectability, detection line does not develop the color, the nature controlling line exhibit red; If nature controlling line does not develop the color, it is invalid then to detect.
Compared with prior art, the invention has the advantages that:
(1) highly sensitive, detectability reach 0.005 mg/L, and reason is: the first, and specificity barcode DNA-nanometer gold-DNAzyme probe is very sensitive to lead ion; The second, reduced the DNAzyme consumption in a large number, only need a small amount of lead ion just can the DNAzyme double-spiral structure be disintegrated, nanometer gold is dropped from nature controlling line, improved detection sensitivity; The 3rd, introduce barcode DNA first, a large amount of barcode DNA are attached to the nanometer gold surface, and nanometer gold is very easily captured by captured dna on detection line, have further improved detection sensitivity; The 4th, the molar absorptivity of nanometer gold is very high, and colour developing is distinct; The 5th, detect and on test strip, carry out, be a colour developing under white background, people's naked eyes are more prone to identification; The 6th, test strip can effectively be eliminated the influence of coloured interfering substance to colour developing through filtering and the expansion effect.
(2) highly selective, common metal ion such as Hg
2+, Cd
2+, Mg
2+, Ca
2+, Fe
2+, Cu
2+, Ni
2+, Co
2+, Mn
2+, Zn
2+All noiseless to detecting.Reason is: the DNAzyme in specificity barcode DNA-nanometer gold-DNAzyme probe has the recognition capability of high specific to lead ion, and the interference of other metals ion can be ignored.
(3) sample need not complicated pre-treatment.Reason is: but specificity barcode DNA-nanometer gold-DNAzyme probe specific recognition lead ion, and can eliminate sample matrices and disturb through the filtration capacity of expansion effect and test strip itself.
(4) simple to operate, detection time is short, only needs several minutes, and reason is: only need can observe detected result through simple immersion, expansion.
(5) the detection cost is low, about 5 yuan of each test strip cost of manufacture, and reason is: reagent and material usage are few, and production stage is simple.
In sum, based on the lead ion test strip of nanometer gold, DNAzyme and barcode DNA, this test strip can highly sensitive, the lead ion in the highly selective test sample, and the preparation method of this test strip is simple, easy to operate.
Description of drawings
Fig. 1 is the structural representation of test strip of the present invention;
Fig. 2 detects the effect synoptic diagram for the present invention;
A: contain the lead ion that is higher than detectability in the sample liquid;
B: not leaded or lead ion content is lower than detectability in the sample liquid;
C: it is invalid to detect;
Fig. 3 detects the figure as a result of actual sample for the present invention.
Embodiment
Embodiment describes in further detail the present invention below in conjunction with accompanying drawing.
Embodiment 1
A kind of lead ion test strip of the present invention; Form successively by the sample pad 2 of overlap joint, the coating specificity barcode DNA-nanometer gold-glass fiber conjugate pad 3 of DNAzyme probe, the nitrocellulose membrane 4 of specified packet quilt and absorbent pad 5 by bar shaped base plate 1 with on bar shaped base plate 1 for this test strip; This specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold; In the nanometer gold surface bonding some barcode DNA are arranged; Useful strepto-affinity element encapsulates orthoscopic nature controlling line C line 6 and encapsulates orthoscopic detection line T line 7, complementary pairing between the base of the barcode DNA of the base of this captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface with biotinylated captured dna and the plain bonded mixture of strepto-affinity on the nitrocellulose membrane
The structure of described specificity barcode DNA-nanometer gold-DNAzyme probe does
, the structure of captured dna is biotin-5'-TTTTTTTTTTTTTTT-3', the diameter of nanometer gold is 13～30 nm.
In this specific embodiment, sample pad 2 is the glass fiber paper through 0.01～0.05 mol/L Tris-HCl damping fluid immersion treatment, and wherein to contain the mass percent of sucrose be 1～10% to the Tris-HCl damping fluid, and the pH value is 5.0～8.0.
In this specific embodiment, the specificity barcode DNA-nanometer gold-quantity for spray of DNAzyme probe solution on glass fiber conjugate pad 3 is 2.0～10 μ L/cm
2The compound method of specificity barcode DNA-nanometer gold-DNAzyme probe solution is following: with 2 μ L concentration is that the sequence of 100 μ M is that to be called the DNA of 17DS1 and sequence that 2 μ L concentration are 100 μ M be to obtain the DNAzyme binding substances after the DNA that is called 17E1 of 5'-TGACATCTCTTCTCCGAGCCGGTCGAAATAGTG-3' mixes for the name of biotin-3'-ACTGTAGAGAAGG rA TATCACAAAAAAAAAAAAAAA-5'-SH; Be that the sequence of 100 μ M is after the barcode DNA of HS-5'-AAAAAAAAAAAAAAA-3' mixes with DNAzyme binding substances and 8 μ L concentration then; The nano-Au solution of 50 nM of 4 mL is added in the said mixture; Behind incubation 16 h; Centrifugal abandoning supernatant; With gained in the Tris-HCl damping fluid that precipitates 25 mM that are dissolved in 50 μ L again; Wherein the Tris-HCl pH of buffer is 8.0, and containing NaCl concentration is 100 mM, and the massfraction of sucrose is 8%.
In this specific embodiment; The plain bonded mixture of biotinylated captured dna and strepto-affinity encapsulates orthoscopic detection line T line, and to prepare process following: the strepto-affinity element of the biotinylated captured dna solution of the 100 μ M of 108 μ L and 2 mg/mL of 50 μ L mixes; Under the room temperature behind incubation 2 h; Centrifugal 10 min under 12000 rpm, abandoning supernatant is after dissolving again with the PBS damping fluid of 25 mM of 54 μ L; The centrifuging and taking supernatant is sprayed at nitrocellulose membrane with 1～3 μ L/cm and forms detection line T line.
In this specific embodiment, it is 1.0～3.0 μ g/cm that strepto-affinity element encapsulates consumption.
The preparation method of a kind of lead ion test strip of the present invention, concrete steps are following:
1, the preparation of sample pad 2
With glass fiber paper with 0.01～0.05 mol/L Tris-HCl damping fluid soak wetting after, promptly obtained sample pad down in dry 4～12 hours at 20 ℃～37 ℃, wherein to contain the massfraction of sucrose be 1～10% to the Tris-HCl damping fluid, the pH value is 5.0～8.0.
2, apply the preparation of the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe
(massfraction that this Tris-HCl damping fluid contains sucrose is 1～10% with 0.01～0.05 mol/L Tris-HCl damping fluid with glass fiber paper; The pH value is 5.0～8.0) soak wetting after; After under 20 ℃～37 ℃ dry 4～12 hours; On every square centimeter glass fiber paper, evenly spray specificity barcode DNA-nanometer gold-DNAzyme probe solution of 2.0～10.0 μ L; After under 20 ℃～37 ℃ dry 2～5 hours, promptly obtain applying the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe, above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold; In the nanometer gold surface bonding barcode DNA is arranged, the structure of this specificity barcode DNA-nanometer gold-DNAzyme probe is with above-mentioned embodiment 1, and the diameter of this nanometer gold is 13～30 nm; The preparation process of above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe solution is following:
A. be the name of the biotin-3'-ACTGTAGAGAAGG rA TATCACAAAAAAAAAAAAAAA-5'-SH DNA that is called 17DS1 with the 100 μ M sequences of 2 μ L and the 100 μ M sequences of 2 μ L are that the DNA that is called 17E1 of 5'-TGACATCTCTTCTCCGAGCCGGTCGAAATAGTG-3' mixes; At 90 C water-baths, 5 min, naturally cool to after the room temperature the DNAzyme binding substances;
B. 100 μ M sequences with above-mentioned DNAzyme binding substances and 8 μ L are after HS-5'-AAAAAAAAAAAAAAA-3' barcode DNA mixes; The 50 nM nano-Au solutions of 4 mL are added in the said mixture, behind incubation 16 h, centrifugal 15 min of 12000 rpm; Abandoning supernatant; Deposition is dissolved in the Tris-HCl damping fluid (pH is 8.0, and containing NaCl concentration is 100 mM, and the massfraction of sucrose is 8%) of 25 mM of 50 μ L again; After mixing, promptly get specificity barcode-DNA-nanometer gold-DNAzyme probe solution.
3, the preparation of the nitrocellulose membrane 4 of specified packet quilt
Using spray appearance appearance is that the strepto-affinity cellulose solution of 1 mg/mL evenly is sprayed on the nitrocellulose membrane 4 with concentration; Drying is 2～8 hours under 25～37 ℃; Form orthoscopic nature controlling line C line 6; With spray appearance appearance the amount edge of the plain bonded complex solution of biotinylated captured dna and strepto-affinity with 1～3 μ L/cm evenly is sprayed on the nitrocellulose membrane 4 with nature controlling line C line 6 parallel directions; Drying is 2～8 hours under 25 ℃～37 ℃, forms orthoscopic detection line T line 7, promptly obtains the nitrocellulose membrane 4 of specified packet quilt; Wherein can form complementary pairing between the base of the barcode DNA of the base of captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of captured dna is biotin-5'-TTTTTTTTTTTTTTT-3'.
It is 1.0～3.0 μ L/cm that above-mentioned strepto-affinity cellulose solution encapsulates consumption; The solvent of this strepto-affinity cellulose solution is concentration 0.01～0.05 a mol/L PBS damping fluid; Containing the NaCl massfraction in this PBS damping fluid is 0.5～1%; Containing the sodium lauryl sulphate massfraction is 0.01～0.05%, and described PBS pH of buffer value is 5.0～8.0.
The plain bonded complex solution of above-mentioned biotinylated captured dna and strepto-affinity process for preparation is following: the strepto-affinity element of the captured dna solution of the 100 μ M of 108 μ L and 2 mg/mL of 50 μ L mixes; Under the room temperature behind incubation 2 h; Centrifugal 10 min under 12000 rpm; Abandoning supernatant after dissolving again with the PBS damping fluid of 25 mM of 54 μ L, promptly obtains the plain bonded complex solution of biotinylated captured dna and strepto-affinity.
4, lead ion test strip preparation
With sample pad 2; Apply the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe; The nitrocellulose membrane 4 and the absorbent pad 5 of specified packet quilt overlap and are pasted on the bar shaped base plate 1 according to the order of sequence; Be cut into the wide slice of 3～5 mm, promptly get lead ion test strip (like Fig. 1, shown in Figure 2).
Embodiment 3
A kind of lead ion test strip of the present invention; Form successively by the sample pad 2 of overlap joint, the coating specificity barcode DNA-nanometer gold-glass fiber conjugate pad 3 of DNAzyme probe, the nitrocellulose membrane 4 of specified packet quilt and absorbent pad 5 by bar shaped base plate 1 with on bar shaped base plate 1 for this test strip; This specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold; In the nanometer gold surface bonding barcode DNA is arranged; Useful strepto-affinity element encapsulates orthoscopic nature controlling line C line 6 and encapsulates orthoscopic detection line T line 7 with biotinylated captured dna and the plain bonded mixture of strepto-affinity on the nitrocellulose membrane; Complementary pairing between the base of the barcode DNA of the base of this captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, its concrete preparation as follows:
1, the preparation of sample pad 2
With glass fiber paper with 0.01 mol/L Tris-HCl damping fluid soak wetting after, promptly obtained sample pad down in dry 4 hours at 20 ℃, wherein to contain the massfraction of sucrose be 1% to the Tris-HCl damping fluid, the pH value is 5.0;
2, apply the preparation of the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe
(massfraction that this Tris-HCl damping fluid contains sucrose is 1% with 0.01 mol/L Tris-HCl damping fluid with glass fiber paper; The pH value is 5.0) soak wetting after; After under 20 ℃ dry 4 hours; On every square centimeter glass fiber paper, evenly spray specificity barcode DNA-nanometer gold-DNAzyme probe solution of 2.0 μ L; After under 20 ℃～37 ℃ dry 2 hours, promptly obtain applying the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe, the structure of this specificity barcode DNA-nanometer gold-DNAzyme probe is with above-mentioned embodiment 1; The diameter of nanometer gold is 13 nm, and the preparation process of above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe solution is with embodiment 2;
3, the preparation of the nitrocellulose membrane 4 of specified packet quilt
Using spray appearance appearance is that the strepto-affinity cellulose solution of 1 mg/mL evenly is sprayed on the nitrocellulose membrane 4 with concentration; Drying is 2 hours under 25 ℃; Form orthoscopic nature controlling line C line 6; With spray appearance appearance the amount edge of the plain bonded complex solution of biotinylated captured dna and strepto-affinity with 1 μ L/cm evenly is sprayed on the nitrocellulose membrane 4 with nature controlling line C line 6 parallel directions, 25 ℃ dry 2 hours down, formation orthoscopic detection line T line 7; Promptly obtain the nitrocellulose membrane 4 of specified packet quilt, wherein the structure of captured dna is biotin-5'-TTTTTTTTTTTTTTT-3';
It is 1.0 μ L/cm that above-mentioned strepto-affinity cellulose solution encapsulates consumption; The solvent of this strepto-affinity cellulose solution is concentration 0.01 a mol/L PBS damping fluid; Containing the NaCl massfraction in this PBS damping fluid is 0.5%; Containing the sodium lauryl sulphate massfraction is 0.01%, and described PBS pH of buffer value is 5.0; The compound method of the plain bonded complex solution of above-mentioned biotinylated captured dna and strepto-affinity is with above-mentioned embodiment 2;
4, lead ion test strip preparation
With sample pad 2; Apply the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe; The nitrocellulose membrane 4 and the absorbent pad 5 of specified packet quilt overlap and are pasted on the bar shaped base plate 1 according to the order of sequence; Be cut into the wide slice of 3 mm, promptly get lead ion test strip (like Fig. 1, shown in Figure 2).
A kind of lead ion test strip of the present invention; Form successively by the sample pad 2 of overlap joint, the coating specificity barcode DNA-nanometer gold-glass fiber conjugate pad 3 of DNAzyme probe, the nitrocellulose membrane 4 of specified packet quilt and absorbent pad 5 by bar shaped base plate 1 with on bar shaped base plate 1 for this test strip; This specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold; In the nanometer gold surface bonding barcode DNA is arranged; Useful strepto-affinity element encapsulates orthoscopic nature controlling line C line 6 and encapsulates orthoscopic detection line T line 7, complementary pairing between the base of the barcode DNA of the base of this captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface with biotinylated captured dna and the plain bonded mixture of strepto-affinity on the nitrocellulose membrane
Its concrete preparation as follows:
1, the preparation of sample pad 2
With glass fiber paper with 0.03 mol/L Tris-HCl damping fluid soak wetting after, promptly obtained sample pad down in dry 8 hours at 30 ℃, wherein to contain the massfraction of sucrose be 5% to the Tris-HCl damping fluid, pH is 6.5;
2, apply the preparation of the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe
(massfraction that this Tris-HCl damping fluid contains sucrose is 5% with 0.03 mol/L Tris-HCl damping fluid with glass fiber paper; The pH value is 6.5) soak wetting after; After under 30 ℃ dry 8 hours; On every square centimeter glass fiber paper, evenly spray specificity barcode DNA-nanometer gold-DNAzyme probe solution of 6 μ L, after under 30 ℃ dry 3.5 hours, promptly obtain applying the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe; Above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold, in the nanometer gold surface bonding barcode DNA is arranged, and the structure of this specificity barcode DNA-nanometer gold-DNAzyme probe is with above-mentioned embodiment 1; The diameter of nanometer gold is 20 nm, and the preparation of above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe solution is with embodiment 2;
3, the preparation of the nitrocellulose membrane 4 of specified packet quilt
Using spray appearance appearance is that the strepto-affinity cellulose solution of 1 mg/mL evenly is sprayed on the nitrocellulose membrane 4 with concentration; Drying is 5 hours under 30 ℃; Form orthoscopic nature controlling line C line 6; With spray appearance appearance the amount edge of the plain bonded complex solution of biotinylated captured dna and strepto-affinity with 2 μ L/cm evenly is sprayed on the nitrocellulose membrane 4 with nature controlling line C line 6 parallel directions; Drying is 5 hours under 30 ℃, forms orthoscopic detection line T line 7, promptly obtains the nitrocellulose membrane 4 of specified packet quilt; Wherein can form complementary pairing between the base of the barcode DNA of the base of captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of captured dna is biotin-5'-TTTTTTTTTTTTTTT-3';
It is 2.0 μ L/cm that above-mentioned strepto-affinity cellulose solution encapsulates consumption; The solvent of this strepto-affinity cellulose solution is concentration 0.03 a mol/L PBS damping fluid; Containing the NaCl massfraction in this PBS damping fluid is 0.8%, and containing the sodium lauryl sulphate massfraction is 0.03%, and described PBS pH of buffer is 6.5; The preparation method of the plain bonded complex solution of above-mentioned biotinylated captured dna and strepto-affinity is with above-mentioned embodiment 2;
4, lead ion test strip preparation
With sample pad 2; Apply the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe; The nitrocellulose membrane 4 and the absorbent pad 5 of specified packet quilt overlap and are pasted on the bar shaped base plate 1 according to the order of sequence; Be cut into the wide slice of 4 mm, promptly get lead ion test strip (like Fig. 1, shown in Figure 2).
Embodiment 5
A kind of lead ion test strip of the present invention; Form successively by the sample pad 2 of overlap joint, the coating specificity barcode DNA-nanometer gold-glass fiber conjugate pad 3 of DNAzyme probe, the nitrocellulose membrane 4 of specified packet quilt and absorbent pad 5 by bar shaped base plate 1 with on bar shaped base plate 1 for this test strip; Useful strepto-affinity element encapsulates orthoscopic nature controlling line C line 6 and encapsulates orthoscopic detection line T line 7 with biotinylated captured dna and the plain bonded mixture of strepto-affinity on the nitrocellulose membrane; Complementary pairing between the base of the barcode DNA of the base of this captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, it is following that it specifically prepares process:
1, the preparation of sample pad 2
With glass fiber paper with 0.05 mol/L Tris-HCl damping fluid soak wetting after, promptly obtained sample pad down in dry 12 hours at 37 ℃, wherein to contain the massfraction of sucrose be 10% to the Tris-HCl damping fluid, the pH value is 8.0;
2, apply the preparation of the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe
With glass fiber paper with 0.05 mol/L Tris-HCl damping fluid soak wetting after; After under 37 ℃ dry 12 hours; On every square centimeter glass fiber paper, evenly spray specificity barcode DNA-nanometer gold-DNAzyme probe solution of 10.0 μ L, after under 37 ℃ dry 5 hours, promptly obtain applying the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe; Above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold, in the nanometer gold surface bonding barcode DNA is arranged, and the structure of this specificity barcode DNA-nanometer gold-DNAzyme probe is with above-mentioned embodiment 1; The diameter of nanometer gold is 30 nm, and the preparation process of above-mentioned specificity barcode DNA-nanometer gold-DNAzyme probe solution is with embodiment 2;
3, the preparation of the nitrocellulose membrane 4 of specified packet quilt
Using spray appearance appearance is that the strepto-affinity cellulose solution of 1 mg/mL evenly is sprayed on the nitrocellulose membrane 4 with concentration; Drying is 8 hours under 37 ℃; Form orthoscopic nature controlling line C line 6; With spray appearance appearance the amount edge of the plain bonded complex solution of biotinylated captured dna and strepto-affinity with 3 μ L/cm evenly is sprayed on the nitrocellulose membrane 4 with nature controlling line C line 6 parallel directions; Drying is 8 hours under 37 ℃, forms orthoscopic detection line T line 7, promptly obtains the nitrocellulose membrane 4 of specified packet quilt; Wherein can form complementary pairing between the base of the barcode DNA of the base of captured dna and specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of captured dna is biotin-5'-TTTTTTTTTTTTTTT-3';
It is 3.0 μ L/cm that above-mentioned strepto-affinity cellulose solution encapsulates consumption; The solvent of this strepto-affinity cellulose solution is concentration 0.05 a mol/L PBS damping fluid; Containing the NaCl massfraction in this PBS damping fluid is 1%, and containing the sodium lauryl sulphate massfraction is 0.05%, and described PBS pH of buffer is 8.0; The preparation method of the plain bonded complex solution of above-mentioned biotinylated captured dna and strepto-affinity is with above-mentioned embodiment 2;
4, lead ion test strip preparation
With sample pad 2; Apply the glass fiber conjugate pad 3 of specificity barcode DNA-nanometer gold-DNAzyme probe; The nitrocellulose membrane 4 and the absorbent pad 5 of specified packet quilt overlap and are pasted on the bar shaped base plate 1 according to the order of sequence; Be cut into the wide slice of 5 mm, promptly get lead ion test strip (like Fig. 1, shown in Figure 2).
The detection test of embodiment 6 highly sensitive and highly selective
According to embodiment 2 preparation lead ion test strip, detect mixing solutions respectively and (contain Hg
2+, Cd
2+, Mg
2+, Ca
2+, Fe
2+, Cu
2+, Ni
2+, Co
2+, Mn
2+, Zn
2+, each ionic concn is 1 mg/L), 0.01 mg/L Pb
2+Solution.This lead ion test strip is inserted mixing solutions and Pb respectively
2+In the solution, the result shown in Fig. 3 (a), Fig. 3 (b), explains that lead ion test strip of the present invention has highly selective and highly sensitive respectively.
The application of test strip in preparation lead ion test card; Test card comprises plastic clip and lead ion test strip, and this plastic clip is a card with plastics preparations, is made up of two getting stuck of can being entrenched togather up and down; Last slice except a hole that drips sample liquid is arranged; Also indicate C, T, T representes the position of detection line, and C representes the position of nature controlling line.Use this lead ion test card only to need in the sample drop hand-hole, this process operation is simple, and detection time is short, only needs several minutes, and it is low to detect cost, about 5 yuan of each test strip cost of manufacture, and sample also need not complicated pre-treatment.
Present embodiment provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited only to the foregoing description, and protection scope of the present invention is as the criterion with claims being to implement under the prerequisite with technical scheme of the present invention.
Claims (10)
1. lead ion test strip; It is characterized in that: form successively by the sample pad of overlap joint, the coating specificity barcode DNA-nanometer gold-glass fiber conjugate pad of DNAzyme probe, the nitrocellulose membrane and the absorbent pad of specified packet quilt by the bar shaped base plate with on the bar shaped base plate for described test strip; Described specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme; Its other end is combined with nanometer gold; In the nanometer gold surface bonding some barcode DNA are arranged; On described nitrocellulose membrane, encapsulate orthoscopic nature controlling line C line with strepto-affinity element, encapsulate orthoscopic detection line T line with biotinylated captured dna and the plain bonded mixture of strepto-affinity; Complementary pairing between the base of the barcode DNA of the base of described biotinylated captured dna and described specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of described specificity barcode DNA-nanometer gold-DNAzyme probe does
, the structure of described biotinylated captured dna is biotin-5'-TTTTTTTTTTTTTTT-3', the diameter of described nanometer gold is 13～30 nm.
2. a kind of lead ion test strip according to claim 1; It is characterized in that: described sample pad is the glass fiber paper through 0.01～0.05 mol/L Tris-HCl damping fluid immersion treatment; Wherein to contain the mass percent of sucrose be 1～10% to the Tris-HCl damping fluid, and the pH value is 5.0～8.0.
3. a kind of lead ion test strip according to claim 1 is characterized in that: the described specificity barcode DNA-nanometer gold-quantity for spray of DNAzyme probe solution on described glass fiber conjugate pad is 2.0～10 μ L/cm
2
4. a kind of lead ion test strip according to claim 3; It is characterized in that: described specificity barcode DNA-nanometer gold-DNAzyme probe solution be the 100 μ M sequences of 2 μ L be the 100 μ M sequences of DNA and the 2 μ L of biotin-3'-ACTGTAGAGAAGG rA TATCACAAAAAAAAAAAAAAA-5'-SH be the DNA of 5'-TGACATCTCTTCTCCGAGCCGGTCGAAATAGTG-3' mix the DNAzyme binding substances; 100 μ M sequences with 8 μ L are after HS-5'-AAAAAAAAAAAAAAA-3' barcode DNA and DNAzyme binding substances mix then; The 50 nM nano-Au solutions of 4 mL are added in the said mixture; Behind incubation 16 h; Centrifugal abandoning supernatant, deposition are dissolved in 50 μ L again and contain gained in the 25 mM Tris-HCl damping fluids of 100 mM NaCl and 8% sucrose.
5. a kind of lead ion test strip according to claim 1; It is characterized in that: the plain bonded mixture of described biotinylated captured dna and strepto-affinity encapsulates orthoscopic detection line T line, and to prepare process following: the strepto-affinity cellulose solution of the biotinylated captured dna solution of the 100 μ M of 108 μ L and 2 mg/mL of 50 μ L mixes; Under the room temperature behind incubation 2 h; Centrifugal 10 min under 12000 rpm; Abandoning supernatant after dissolving again with the PBS damping fluid of 25 mM of 54 μ L, is sprayed at nitrocellulose membrane with 1～3 μ L/cm and forms detection line T line.
6. a kind of lead ion test strip according to claim 1 is characterized in that: it is 1.0～3.0 μ g/cm that described strepto-affinity element encapsulates consumption.
7. the preparation method of a lead ion test strip according to claim 1 is characterized in that may further comprise the steps:
(1) preparation of sample pad
With glass fiber paper with 0.01～0.05 mol/L Tris-HCl damping fluid soak wetting after, promptly obtained sample pad down in dry 4～12 hours at 20 ℃～37 ℃, wherein to contain the massfraction of sucrose be 1～10% to the Tris-HCl damping fluid, the pH value is 5.0～8.0;
(2) preparation of the glass fiber conjugate pad of coating specificity barcode DNA-nanometer gold-DNAzyme probe
With glass fiber paper with 0.01～0.05 mol/L Tris-HCl damping fluid soak wetting after; After under 20 ℃～37 ℃ dry 4～12 hours; On every square centimeter glass fiber paper, evenly spray specificity barcode DNA-nanometer gold-DNAzyme probe solution of 2.0～10.0 μ L; After under 20 ℃～37 ℃ dry 2～5 hours; Promptly obtain applying the glass fiber conjugate pad of specificity barcode DNA-nanometer gold-DNAzyme probe, described specificity barcode DNA-nanometer gold-DNAzyme probe is combined with vitamin H at an end of DNAzyme, and its other end is combined with nanometer gold; In the nanometer gold surface bonding some barcode DNA are arranged, the structure of said specificity barcode DNA-nanometer gold-DNAzyme probe does
, wherein to contain the mass percent of sucrose be 1～10% to the Tris-HCl damping fluid, and the pH value is 5.0～8.0, and the diameter of described nanometer gold is 13～30 nm;
(3) preparation of the nitrocellulose membrane of specified packet quilt
Using spray appearance appearance is that the strepto-affinity cellulose solution of 1 mg/mL evenly is sprayed on the nitrocellulose membrane with concentration; Drying is 2～8 hours under 25 ℃～37 ℃; Form orthoscopic nature controlling line C line; Evenly be sprayed at nitrocellulose membrane on the direction of the amount edge of 1～3 μ L/cm and nature controlling line C line parallel biotinylated captured dna and strepto-affinity element bonded complex solution with spray appearance appearance; Drying is 2～8 hours under 25 ℃～37 ℃, forms orthoscopic detection line T line, promptly obtains the nitrocellulose membrane of specified packet quilt; Can form complementary pairing between the base of the barcode DNA of the base of wherein said biotinylated captured dna and described specificity barcode DNA-nanometer gold-DNAzyme detecting probe surface, the structure of described barcode DNA is biotin-5'-TTTTTTTTTTTTTTT-3';
(4) sample pad that step (1) is obtained; The glass fiber conjugate pad of coating specificity barcode DNA-nanometer gold-DNAzyme probe that step (2) obtains; The nitrocellulose membrane of the specified packet quilt that step (3) obtains and absorbent pad overlap according to the order of sequence and are pasted on the base plate; Be cut into the wide slice of 3～5 mm, promptly get the lead ion test strip.
8. the preparation method of a kind of lead ion test strip according to claim 7; It is characterized in that: the preparation method of described specificity barcode DNA-nanometer gold-DNAzyme probe solution is following: the 100 μ M sequences of 2 μ L are that the 100 μ M sequences of DNA and the 2 μ L of biotin-3'-ACTGTAGAGAAGG rA TATCACAAAAAAAAAAAAAAA-5'-SH are that the DNA of 5'-TGACATCTCTTCTCCGAGCCGGTCGAAATAGTG-3' mixes; At 90 C water-baths, 5 min; Get the DNAzyme binding substances after naturally cooling to room temperature; Be after barcode DNA and the DNAzyme binding substances of HS-5'-AAAAAAAAAAAAAAA-3' mixes with the 100 μ M sequences of 8 μ L then, the 50 nM nano-Au solutions of 4 mL are added in the said mixture, behind incubation 16 h; Centrifugal 15 min of 12000 rpm; Abandoning supernatant, deposition is dissolved in 50 μ L again and contains in 100 mM NaCl and the 25 mM Tris-HCl damping fluids, after mixing; Promptly get specificity barcode DNA-nanometer gold-DNAzyme probe solution; The mass percent that wherein said Tris-HCl damping fluid contains sucrose is 1～10%, and the concentration that contains NaCl is 100 mM, and described Tris-HCl pH of buffer value is 5.0～8.0.
9. the preparation method of a kind of lead ion test strip according to claim 7; It is characterized in that: it is 1.0～3.0 μ L/cm that described strepto-affinity cellulose solution encapsulates consumption; The solvent of described strepto-affinity cellulose solution is concentration 0.01～0.05 a mol/L PBS damping fluid; Containing the NaCl mass percent in the described PBS damping fluid is 0.5～1%, and containing the sodium lauryl sulphate mass percent is 0.01～0.05%, and described PBS pH of buffer value is 5.0～8.0.
10. the preparation method of a kind of lead ion test strip according to claim 7; It is characterized in that: the plain bonded complex solution of described biotinylated captured dna and strepto-affinity process for preparation is following: the strepto-affinity element of the captured dna solution of the 100 μ M of 108 μ L and 2 mg/mL of 50 μ L mixes; Under the room temperature behind incubation 2 h; Centrifugal 10 min under 12000 rpm; Abandoning supernatant after dissolving again with the PBS damping fluid of 25 mM of 54 μ L, promptly obtains the plain bonded complex solution of biotinylated captured dna and strepto-affinity.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103852436A (en) * | 2014-03-14 | 2014-06-11 | 厦门大学 | Method for detecting lead ion through high-specificity DNA hydrogel |
| CN104155290A (en) * | 2014-06-25 | 2014-11-19 | 宁波大学 | Preparation method for solid electrochemical luminescence sensor for detecting lead ions and application of solid electrochemical luminescence sensor for detecting lead ions |
| CN105181947A (en) * | 2015-08-20 | 2015-12-23 | 中国科学院广州生物医药与健康研究院 | G-tetramer based detection method |
| CN110672834A (en) * | 2019-09-26 | 2020-01-10 | 北京丹大生物技术有限公司 | Kit for rapidly detecting lead content in sample |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253213A (en) * | 2011-07-06 | 2011-11-23 | 江南大学 | Colloidal gold chromatography test strip for quickly detecting lead ions |
| CN102352413A (en) * | 2011-10-17 | 2012-02-15 | 中国科学院广州生物医药与健康研究院 | Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof |
| CN102426239A (en) * | 2011-09-16 | 2012-04-25 | 王利兵 | Colloidal gold chromatographic test strip for rapidly testing lead ion |
-
2012
- 2012-04-25 CN CN2012101247360A patent/CN102643917B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253213A (en) * | 2011-07-06 | 2011-11-23 | 江南大学 | Colloidal gold chromatography test strip for quickly detecting lead ions |
| CN102426239A (en) * | 2011-09-16 | 2012-04-25 | 王利兵 | Colloidal gold chromatographic test strip for rapidly testing lead ion |
| CN102352413A (en) * | 2011-10-17 | 2012-02-15 | 中国科学院广州生物医药与健康研究院 | Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103852436A (en) * | 2014-03-14 | 2014-06-11 | 厦门大学 | Method for detecting lead ion through high-specificity DNA hydrogel |
| CN103852436B (en) * | 2014-03-14 | 2016-08-17 | 厦门大学 | A kind of high specific DNA hydrogel detection method to lead ion |
| CN104155290A (en) * | 2014-06-25 | 2014-11-19 | 宁波大学 | Preparation method for solid electrochemical luminescence sensor for detecting lead ions and application of solid electrochemical luminescence sensor for detecting lead ions |
| CN105181947A (en) * | 2015-08-20 | 2015-12-23 | 中国科学院广州生物医药与健康研究院 | G-tetramer based detection method |
| CN110672834A (en) * | 2019-09-26 | 2020-01-10 | 北京丹大生物技术有限公司 | Kit for rapidly detecting lead content in sample |
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