CN112698020A - Multimodal coupling analysis method of cross response system based on DNA-AuNP coding - Google Patents
Multimodal coupling analysis method of cross response system based on DNA-AuNP coding Download PDFInfo
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Abstract
The invention belongs to the field of biomedical sensing detection, and particularly relates to a multimodal coupling analysis method of a cross-response sensing system based on DNA-AuNP coding, which comprises the following steps: modifying the gold nanoparticles by polyA-DNA probes with different lengths to obtain various DNA-AuNPs with different binding capacities of polyA-DNA and the surfaces of the gold nanoparticles, and completing the construction of a cross-response sensor taking the aggregation degree of the polyA-DNA modified gold nanoparticles with different lengths as a variation factor; obtaining fingerprint information of aggregation degrees of different types of gold nanoparticles induced by the object to be detected by coupling multimodal signals; and classifying and identifying different types of objects to be detected according to the fingerprint information. The invention can solve the technical problems of complicated biochemical detection and classification process, strong specialization, high cost and easy interference of the existing metal ions, and provides an analysis method with convenient detection, time saving, non-specificity and high sensitivity. The invention can also detect and compare the fingerprint information obtained by the body fluids of the normal person and the patient, construct an aggregation degree database and hopefully improve the distinction between the normal person and the patient.
Description
Technical Field
The invention belongs to the field of biomedical sensing detection, and particularly relates to a multimodal coupling analysis method of a cross response system based on DNA-AuNP coding.
Background
The gold nanoparticle compound is widely applied to the field of biomedical sensing detection, but most of the existing characterization methods only consider the single modal characteristic of the reaction of the single gold nanoparticle compound caused by a detection object, and ignore the complementary advantages among different modal characteristics; meanwhile, in the data processing process, the same weight is usually given to all data, wherein the quality of the extracted features is affected by the information of weak correlation, so that the detection of the detected object with high precision and high sensitivity cannot be realized.
For example: the current common gold nanoparticle composite analysis method comprises the following steps: ultraviolet spectrophotometry (UV), which utilizes electronic resonance on the surface of gold nanoparticles, and gathers gold nanoparticles to different degrees when an object to be detected interacts with modifiers on the surface of the gold nanoparticles, thereby obtaining ultraviolet absorption spectrum fingerprint data, wherein the single-mode analysis method has low accuracy and low detection efficiency due to the influence of the number of specific receptors; atomic Absorption Spectroscopy (AAS), inductively coupled plasma mass spectrometry (ICP-MS), Surface Enhanced Raman (SERS), electron microscopy (SEM, TEM), etc., have been greatly limited in their application due to the high cost and complex sample pretreatment. Based on these problems, it is important to develop a cross-response sensor that can analyze a plurality of signals quickly and has a simple detection means.
Disclosure of Invention
The invention provides a multimodal coupling analysis method of a cross response system based on DNA-AuNP coding, which can realize high-precision and high-sensitivity classification of various objects to be detected, and aims to overcome the problem that the existing detection method can not realize high-precision and high-sensitivity detection.
In order to solve the technical problems, the technical scheme of the invention is as follows: a multimodal coupling analysis method based on a cross-response system encoded by DNA-AuNP, said method comprising the steps of:
modifying the gold nanoparticles by using polyA-DNA probes with different lengths to obtain various DNA-AuNPs with different binding capacities of the polyA-DNA and the gold nanoparticles, wherein the different DNA-AuNPs have different aggregation degrees on a substance to be detected; thus completing the construction of the cross-response sensor taking the aggregation degree of the polyA-DNA modified gold nanoparticles with different lengths as a change factor;
obtaining fingerprint information of aggregation degrees of different types of gold nanoparticles induced by the object to be detected by coupling multimodal signals; and classifying and identifying different types of objects to be detected according to the fingerprint information.
Preferably, the analyte inducing the gold nanoparticles to aggregate includes, but is not limited to, metal ions, bacteria, body fluids; wherein, the metal ions include but are not limited to: ca2+、Cu2+、Fe3+、Mg2+、Zn2+(ii) a The body fluid includes but is not limited to cerebrospinal fluid, saliva, urine, sweat.
Further, the degree of aggregation of gold nanoparticles yields a multimodal signal by a variety of characterization means including, but not limited to: ultraviolet absorption spectrum, surface zeta potential and particle size.
Still further, the number of adenine bases of the polyA-DNA probe is: 5. 10, 20, 30, 40, 50.
And further, classifying and identifying the obtained fingerprint data by adopting a linear discriminant analysis algorithm.
Further, modifying the gold nanoparticles by using a polyA-DNA probe, which comprises the following steps:
mixing polyA-DNA with the volume of 1uL and the concentration of 100uM with gold nanoparticle solution with the volume of 10uL and the concentration of 100nM, adding Citrate-HCl with the volume of 0.2uL, the concentration of 500mM and the pH of 3, and incubating for 10 minutes at room temperature; and (3) removing the supernatant through centrifugation, adding the supernatant into PBS buffer solution with the volume of 100uL, the concentration of 10mM and the pH of 7, repeating for many times, removing redundant polyA-DNA chains, and finishing modification of the gold nanoparticles by the polyA-DNA probe, wherein the obtained solution is the DNA functionalized gold nanoparticles with the concentration of 10 nM.
Still further, the rotation speed of centrifugation is 14000rpm, and the centrifugation time is 20 min.
Further, a cross-response sensor with the aggregation degree of the gold nanoparticles modified by the polyA-DNA with different lengths as a variation factor is constructed, and the specific steps are as follows:
adding 10uL of polyA-DNA modified gold nanoparticle solution into metal cation solutions with different concentrations and a volume of 90uL to ensure that the final concentration of the polyA-DNA modified gold nanoparticle solution in a reaction system is about 1nM, and carrying out a light-shielding reaction at room temperature;
after 3 hours, the corresponding absorption spectrum is recorded, and the response of the spectrum signal is represented by A/A0Obtaining wherein A and A0Respectively, is UV absorption, A/A0OD626nm/OD524 nm; and simultaneously recording zeta potential and particle size of the aggregated gold nanoparticles, thereby obtaining 'fingerprint' information of ultraviolet fluorescence absorption spectrum, particle size and zeta potential diagram of the DNA-AuNPs system in the presence of different types of metal ions.
Still further, in order to eliminate background interference, the ultraviolet absorption spectrum A/A of gold nanoparticles modified with polyA5-DNA in the absence of metal ions was measured0The resulting data of (a) is normalized.
Compared with the prior art, the technical scheme of the invention has the beneficial effects that:
the method can be used for accurately identifying various metal ions. The DNA-AuNP gold nanoparticles in the system can be aggregated in different degrees due to different concentrations and types of metal ions, and different signals are generated by the ultraviolet absorption spectrum, the surface potential and the particle size of the gold nanoparticles in the cross response sensor, so that the simultaneous high-precision detection and identification of multiple types of metal ions can be realized in a multimodal output signal mode.
The invention takes ultraviolet absorption spectrum, surface potential, particle size and polyA-DNA probes with different lengths as characteristic variable regulating and controlling factors, can meet the requirement of detecting various metal ions by increasing the dimensionality of the characteristic variables, obtains the big data of aggregation degree characteristic signals, and completes artificial intelligent mode recognition training operation and classification operation.
The invention can also detect and compare the fingerprint information obtained by the body fluids of the normal person and the patient, construct an aggregation degree database and hopefully improve the distinction between the normal person and the patient.
Drawings
FIG. 1 is the basic principle of the multimodal coupling analysis method of the DNA-AuNP-encoded cross-response sensor of the embodiment.
FIG. 2 is a diagram showing the preparation of DNA-AuNPs with different lengths and the multi-modal characterization of different aggregation degrees induced by different metal ions in this example.
Wherein, a) a DNA-attenuated coulombic self-assembly process; b) characterizing the AuNPs particle size detected by a Zetasizer Nano particle size analyzer and a Transmission Electron Microscope (TEM); c) ultraviolet absorption spectra of gold nanoparticles modified by polyA-DNA with different lengths; d) difference of ratio (A626/A524) of absorption spectra of polyA-DNA modified gold nanoparticles with different lengths; e) three different modalities, such as Abs; particle size; the surface zeta potential characterizes the aggregation behavior of 5 different DNA-AuNPs in the presence of different metal ions respectively.
FIG. 3 is the "fingerprint" information of the ultraviolet fluorescence absorption spectrum, particle size, zeta potential diagram of the aggregation degree of the gold nanoparticles of A5-AuNPs induced by five different metal ions in this example.
FIG. 4 is a classification diagram and a comparison diagram of identification accuracy of 5 different metal ions by controlling different types of DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs and A50-AuNPs) and numbers under the output of single-mode signals of ultraviolet absorption spectrum (UV-VIS) of the DNA-AuNP-encoded cross-response sensor constructed in this embodiment.
Fig. 5 is a classification diagram and a comparison diagram of identification accuracy of 5 different metal ions by controlling different types of DNA-AuNPs (e.g., a5-AuNPs, a10-AuNPs, a20-AuNPs, a30-AuNPs, a40-AuNPs, and a50-AuNPs) and numbers of the DNA-AuNPs encoded cross-response sensor constructed in this example under the output of a particle size (DLS) single-mode signal.
FIG. 6 is a classification chart and identification accuracy comparison chart of 5 different metal ions by controlling different types of DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs and A50-AuNPs) and numbers under the surface zeta potential (zeta potential chart) single-mode signal output of the DNA-AuNP coding cross-response sensor constructed in this example.
FIG. 7 is a classification chart and a comparison chart of identification accuracy of 5 different metal ions under ultraviolet absorption spectrum (UV-VIS) -particle size (DLS) bimodal signal output of the DNA-AuNP-encoded cross-response sensor constructed in the embodiment and under the participation of 6 DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs and A50-AuNPs).
FIG. 8 is a classification chart and a comparison chart of identification accuracy of 5 different metal ions by 6 DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs and A50-AuNPs) under the output of ultraviolet absorption spectrum (UV-VIS) -surface zeta potential (zeta potential chart) bimodal signals constructed in this example.
FIG. 9 is a classification chart and a comparison chart of identification accuracy of 5 different metal ions by 6 DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs and A50-AuNPs) under the bimodal signal output of particle size (DLS) -surface zeta potential (zeta potential chart) constructed in this example.
FIG. 10 is a classification chart and a comparison chart of identification accuracy of 5 different metal ions by the DNA-AuNP-encoded cross-response sensor constructed in this example under the output of ultraviolet absorption spectrum (UV-VIS) -particle size (DLS) -surface zeta potential (zeta potential chart) tri-modal signals and with the participation of 6 DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs, A50-AuNPs).
FIG. 11 is a classification chart of 5 different metal ions in cerebrospinal fluid, saliva, urine and sweat in 6 DNA-AuNPs (such as A5-AuNPs, A10-AuNPs, A20-AuNPs, A30-AuNPs, A40-AuNPs and A50-AuNPs) under the output of ultraviolet absorption spectrum (UV-VIS) -particle size (DLS) -surface zeta potential (zeta potential diagram) three-mode signals.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and are used for illustration only, and should not be construed as limiting the patent. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The technical solution of the present invention is further described below with reference to the accompanying drawings and examples.
Example 1
As shown in fig. 1 and fig. 2, a multimodal coupling analysis method based on a DNA-AuNP encoded cross response system comprises the following steps:
modifying the gold nanoparticles by using polyA-DNA probes with different lengths to obtain various DNA-AuNPs with different binding capacities of the polyA-DNA and the gold nanoparticles, wherein the different DNA-AuNPs have different aggregation degrees on a substance to be detected; thus completing the construction of the cross-response sensor taking the aggregation degree of the polyA-DNA modified gold nanoparticles with different lengths as a change factor;
obtaining fingerprint information of aggregation degrees of different types of gold nanoparticles induced by the object to be detected by coupling multimodal signals; and classifying and identifying different types of objects to be detected according to the fingerprint information.
The analyte inducing the gold nanoparticles to aggregate comprises but is not limited to metal ions, bacteria and body fluid; wherein, the metal ions include but are not limited to: ca2+、Cu2+、Fe3+、Mg2+、Zn2+(ii) a The body fluid includes but is not limited to cerebrospinal fluid, saliva, urine, sweat.
In the embodiment, the gold nanoparticles are modified by adopting the polyA-DNA probes with different lengths, and different induction factors enable DNA with different lengths to generate competitive reaction with the gold nanoparticles, so that DNA with different lengths can be separated from the surface of the gold nanoparticles to different degrees, thereby inducing the gold nanoparticles to gather, and fingerprint information can be obtained by coupling DNA with different lengths and outputting three signals. Different DNA-AuNPs have different aggregation degrees on metal ions, and according to the characteristics, the method can classify and identify the metal ions and can detect bacteria and body fluid. Any factor that can induce aggregation of gold nanoparticles can be detected using the method described in this example. In this embodiment, the method is described in detail by using gold nanoparticles with a particle size of 13nm, wherein the step of synthesizing the gold nanoparticles with a particle size of 13nm specifically comprises the following steps:
1% trisodium citrate (3.5mL) was added to a boiling and rapidly stirring aqueous solution of HAuCl4(100mL, 0.01%), the solution was kept boiling for 20 minutes, then cooled to room temperature to obtain a gold nanoparticle solution, which was stored at 4 ℃ until use.
In a specific embodiment, the number of bases adenine (A) of the polyA-DNA probe is: 5. 10, 20, 30, 40, 50. The polyA-DNA probe sequences used for modification were as follows:
in a specific example, gold nanoparticles modified with polyA-DNA probes were as follows: mixing polyA-DNA with the volume of 1uL and the concentration of 100uM with gold nanoparticle solution with the volume of 10uL and the concentration of 100nM, adding Citrate-HCl with the volume of 0.2uL, the concentration of 500mM and the pH of 3, and incubating for 10 minutes at room temperature; the supernatant was removed by centrifugation, and added to PBS buffer solution with a volume of 100uL, a concentration of 10mM, and a pH of 7, and repeated several times to remove excess polyA-DNA strands, thereby completing modification of gold nanoparticles with polyA-DNA probe, and the obtained solution was DNA functionalized gold nanoparticles with a concentration of 10nM, which was defined as DNA-AuNP in this example. The rotation speed adopted by the centrifugation is 14000rpm, and the centrifugation time is 20 min.
In a specific embodiment, the metal ion is used as a factor for inducing aggregation of DNA-AuNP, and the method is implementedExamples of metal ions include, but are not limited to: ca2+、Cu2+、Fe3+、Mg2+、Zn2+。
In a specific embodiment, the degree of aggregation of the gold nanoparticles is obtained by a number of characterization means including, but not limited to: ultraviolet absorption spectrum, surface zeta potential and particle size.
In a specific embodiment, a cross-response sensor with the aggregation degree of gold nanoparticles modified by polyA-DNA probes with different lengths as a change factor is constructed, and the specific steps are as follows: 10uL of the AuNPs solution modified by the polyA-DNA is added into metal cation solutions (90uL) with different concentrations, so that the final concentration of the AuNPs solution modified by the polyA-DNA in a reaction system is about 1nM, and the light-shielding reaction is carried out at room temperature. After 3 hours, the corresponding absorption spectra were recorded using a microplate reader and an ultraviolet-visible spectrophotometer. The response of the spectral signal is represented by A/A0Obtaining wherein A and A0Respectively, ultraviolet absorption (a/a0 ═ OD626nm/OD524 nm). To eliminate background interference, the raw data were obtained by modifying AuNPs UV absorption Spectroscopy A/A with polyA5-DNA in the absence of metal ions0The results of (a) are normalized. At the same time, the zeta potential and the particle size of the aggregated nanoparticles were recorded using a Malvern nano-potentiostat ZS 90. Thus, the 'fingerprint data' information of the ultraviolet fluorescence absorption spectrum, the particle size and the zeta potential diagram of the DNA-AuNPs system in the presence of different metal ions is obtained. This example was performed in 8 replicate experiments for each metal ion.
In a specific embodiment, a Linear Discriminant Analysis (LDA) algorithm is used to realize high-precision classification of different types of metal ions according to the obtained "fingerprint data": the method can be specifically operated in the Rstudio, and specific codes are as follows:
setwd ("E:/test") # sets the D-disk down test folder as the working folder, and puts in the csv format data file, where
setwd("E:/test")
library(devtools)
library(ggord)
library(MASS)
library(ggplot2)
data<-read.csv("metal ions.csv",sep＝",",header＝T)
data$group<-as.factor(data$group)
model1＝lda(group～.,data＝data)
model1
plot(model1,dimen＝2)
ld<-predict(model1)$x
ord<-lda(group～.,data)
p < -ggord (ord, data group, arrow ═ 0, vec _ ext ═ 0, size ═ 5, txt ═ NULL, poly ═ FALSE, and coord _ fix ═ F) # drawing
p+theme(panel.grid＝element_blank())
The ultraviolet fluorescence absorption spectrum, the particle size and the fingerprint information of a zeta potential diagram of the DNA-AuNPs system in the presence of five metal ions are shown in figure 3.
In the embodiment, the controllable gold nanoparticles modified by the length of the polyA-DNA sequence are aggregated, and a multi-modal analysis mode recognition algorithm is adopted to achieve simple and high-speed accurate analysis. Compared with the prior traditional DNA-AuNP sensor, the method is limited by the number and the type of receptors. The method improves the condition that the quantity and the type of the recognition receptors are limited due to steric hindrance of the gold nanoparticles, reduces the complexity of a mode recognition algorithm by combining a multi-mode coupling analysis algorithm, and effectively improves the recognition accuracy of rapid detection and accurate analysis. And the method is used for detecting various metal ions, and the accuracy can reach 100%.
In addition, the detection object of the cross-response sensor has universality, and by designing different gold nanoparticles modified by polyA-DNA, rapid and ultrasensitive detection and analysis on different detection objects (including biological small molecules (such as metal ions and the like), biological macromolecules (such as DNA, miRNA, protein and the like), viruses, bacteria, cells and the like) can be realized.
To further illustrate the effect obtained by the method of this embodiment, the following is a demonstration of the classification effect of different metal ions as the number of modes increases, and as the result shows, the classification effect of metal ions becomes better and better as the number of modes increases. Therefore, the fingerprint information generated by different metal ions by adopting the linear discriminant analysis algorithm is practical and effective.
In a specific embodiment, since the body fluid contains metal ions, the method described in the above embodiment can realize accurate distinction of metal ions in different body fluids, specifically as follows:
dissolving metal ions into cerebrospinal fluid, saliva, urine and sweat, and then adding 10ul of polyA-DNA modified gold nanoparticle solution to ensure that the final concentration of the polyA-DNA modified gold nanoparticle solution in the reaction system is about 1nM, and the final concentration of the metal ions is 1 MuM to carry out light-shielding reaction at room temperature; after 3 hours, the corresponding absorption spectrum is recorded, and the response of the spectrum signal is represented by A/A0Obtaining wherein A and A0Respectively, is UV absorption, A/A0OD626nm/OD524 nm; and simultaneously recording zeta potential and particle size of the aggregated gold nanoparticles, thereby obtaining 'fingerprint' information of ultraviolet fluorescence absorption spectrum, particle size and zeta potential diagram of the DNA-AuNPs system in the presence of different types of metal ions. The results thus obtained are shown in fig. 11, from which it can be seen that the metal ion classification can be significantly improved by the method described in this example from fig. 11. Biological fluids including saliva, urine, sweat, and cerebrospinal fluid play an important role in human health. Biological fluids can be analyzed in medical laboratories for the discovery of microorganisms, inflammation, cancer, and the like. Medically, it is a specimen used in diagnostic examinations or evaluations and to determine a disease or condition.
It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (9)
1. A multimodal coupling analysis method based on a DNA-AuNP coding cross response system is characterized by comprising the following steps: the method comprises the following steps:
modifying the gold nanoparticles by using polyA-DNA probes with different lengths to obtain various DNA-AuNPs with different binding capacities of the polyA-DNA and the gold nanoparticles, wherein the different DNA-AuNPs have different aggregation degrees on a substance to be detected; thus completing the construction of the cross-response sensor taking the aggregation degree of the polyA-DNA modified gold nanoparticles with different lengths as a change factor;
obtaining fingerprint information of aggregation degrees of different types of gold nanoparticles induced by the object to be detected by coupling multimodal signals; and classifying and identifying different types of objects to be detected according to the fingerprint information.
2. The method of claim 1 for multimodal coupling analysis based on a cross-response system encoded by DNA-AuNP, characterized in that: the analyte inducing the gold nanoparticles to aggregate comprises but is not limited to metal ions, bacteria and body fluid; wherein, the metal ions include but are not limited to: ca2+、Cu2+、Fe3+、Mg2+、Zn2+(ii) a The body fluid includes but is not limited to cerebrospinal fluid, saliva, urine, sweat.
3. The method of claim 2 for multimodal coupling analysis based on a cross-response system with DNA-AuNP coding, characterized in that: the degree of aggregation of gold nanoparticles a multimodal signal was obtained by a variety of characterization means including, but not limited to: ultraviolet absorption spectrum, surface zeta potential and particle size.
4. The method of claim 3 for multimodal coupling analysis based on a DNA-AuNP-encoded cross-response system, wherein: the number of adenine bases of the polyA-DNA probe is respectively as follows: 5. 10, 20, 30, 40, 50.
5. The method of claim 4 for multimodal coupling analysis based on a DNA-AuNP-encoded cross-response system, wherein: and classifying and identifying the obtained fingerprint data by adopting a linear discriminant analysis algorithm.
6. The method of claim 5 for multimodal coupling analysis based on a DNA-AuNP-encoded cross-response system, wherein: modifying the gold nanoparticles by adopting a polyA-DNA probe, which comprises the following steps:
mixing polyA-DNA with the volume of 1uL and the concentration of 100uM with gold nanoparticle solution with the volume of 10uL and the concentration of 100nM, adding Citrate-HCl with the volume of 0.2uL, the concentration of 500mM and the pH of 3, and incubating for 10 minutes at room temperature; and (3) removing the supernatant through centrifugation, adding the supernatant into PBS buffer solution with the volume of 100uL, the concentration of 10mM and the pH of 7, repeating for many times, removing redundant polyA-DNA chains, and finishing modification of the gold nanoparticles by the polyA-DNA probe, wherein the obtained solution is the DNA functionalized gold nanoparticles with the concentration of 10 nM.
7. The method of claim 6 for multimodal coupling analysis based on a cross-response system encoded by DNA-AuNP, wherein: the rotation speed adopted by the centrifugation is 14000rpm, and the centrifugation time is 20 min.
8. The method of claim 7, wherein the method comprises the following steps: constructing a cross-response sensor taking the aggregation degree of the gold nanoparticles modified by polyA-DNA with different lengths as a variation factor, which comprises the following steps:
adding 10uL of polyA-DNA modified gold nanoparticle solution into metal cation solutions with different concentrations and a volume of 90uL to ensure that the final concentration of the polyA-DNA modified gold nanoparticle solution in a reaction system is 1nM, and carrying out a light-shielding reaction at room temperature;
after 3 hours, recording the corresponding absorption spectrum and the sound of the spectrum signalShall be formed by A/A0Obtaining wherein A and A0Respectively, is UV absorption, A/A0OD626nm/OD524 nm; and simultaneously recording zeta potential and particle size of the aggregated gold nanoparticles, thereby obtaining 'fingerprint' information of ultraviolet fluorescence absorption spectrum, particle size and zeta potential diagram of the DNA-AuNPs system in the presence of different types of metal ions.
9. The method of claim 8 for multimodal coupling analysis based on a cross-response system encoded by DNA-AuNP, characterized in that: to eliminate background interference, the ultraviolet absorption spectrum A/A of gold nanoparticles modified with polyA5-DNA in the absence of metal ions0The resulting data of (a) is normalized.
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