CN108844951A - A kind of mercury ion detecting product, method and smart phone Image analysis system - Google Patents
A kind of mercury ion detecting product, method and smart phone Image analysis system Download PDFInfo
- Publication number
- CN108844951A CN108844951A CN201810688361.8A CN201810688361A CN108844951A CN 108844951 A CN108844951 A CN 108844951A CN 201810688361 A CN201810688361 A CN 201810688361A CN 108844951 A CN108844951 A CN 108844951A
- Authority
- CN
- China
- Prior art keywords
- value
- gray
- image
- detection
- seq
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
- G01N21/8483—Investigating reagent band
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
- G01N21/8483—Investigating reagent band
- G01N2021/8494—Measuring or storing parameters of the band
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides a kind of mercury ion detecting product, method and smart phone Image analysis systems.Sidestream chromatography sensor provided by the invention based on nucleic acid base mispairing includes SEQ ID № in sequence table：At least one nucleotide sequence shown in 1-3, the specific good, high sensitivity of detection.The smart phone Image analysis system that the cooperation Sidestream chromatography sensor provided by the invention uses, the testing result shown on Sidestream chromatography sensor can be converted to the concentration value of mercury ion by mobile phone, what is be simple and efficient realizes the quantitative detection of mercury ion in sample.Mercury ion detecting product, method and smart phone Image analysis system provided by the invention are very suitable for untrained personnel and carry out on-the-spot test, provide great convenience for on-site tests such as food safety, Environmental securities.
Description
Technical field
The invention belongs to technical field of biological, and in particular to a kind of mercury ion detecting product, method and smart phone
Image analysis system.
Background technique
Watersoluble divalent mercury ion is heavy metal classes risks and assumptions relatively conventional in food safety and drinking water safety, tool
There is stronger bioconcentration, it is larger to the harm of human body, nervous system can be damaged in the case where extremely low concentration, disappeared
Change system, brain tissue and nephridial tissue.Many countries and tissue have adjusted the maximum allowable upper limit for drinking mercury ion in water sample, example
As the World Health Organization (World Health Organization, WHO) provides the maximum allowable limitation of drinking water mercury ion not
More than 6ng mL-1(30nM), Environmental Protection Agency USA (Environmental Protec tion Agency, EPA) regulation drink
Acceptable limitation with mercury ion in water is 2ng mL-1(10nM), European Union (European Union, EU) drinking water standard are in
The Ministry of Public Health of state all provides that the highest Permissible limit of mercury ion is no more than 1ng mL-1(5nM).Therefore, the detection of trace amount mercury ion is
The problem of global common concern.Currently, being mainstream based on " nucleic acid base mispairing " identification system construction sensor detection mercury ion
Research tendency, refer to DNA two thymidine alkali can with mispairing combine a mercury ion form stable " T-Hg (II)-
T " structure.But these sensors are faced with the awkward situation that system is complex, is not easy quantitative detection mostly.
For Sidestream chromatography sensor as a kind of emerging quick detection platform, it is quick, simple, single-minded, accurate and clever to possess
The features such as quick.But current Sidestream chromatography sensor can only realize qualitative or half-quantitative detection, and when quantitative detection needs additional
Special instrument, and there is presently no one kind, and quantitative detection data can be simply and easily directly read from Sidestream chromatography sensor
Technology or portable instrument.
Summary of the invention
The present invention provides a kind of mercury ion detecting product, method and smart phone Image analysis systems, can pass through hand
The testing result shown on Sidestream chromatography sensor is converted to the concentration value of mercury ion by machine, is simple and efficient at least realizing
The concentration of mercury ion in quantitative detection sample.Sidestream chromatography sensor provided by the invention based on nucleic acid base mispairing and/or
The smart phone Image analysis system being used cooperatively is very suitable for untrained personnel and carries out on-the-spot test, for food
The on-site tests such as safety, Environmental security provide great convenience.
At least one of it is an object of the present invention to provide composition, composition includes following 1) -3)：
1) SEQ ID № in sequence table：Nucleotide sequence shown in 1；Or by SEQ ID № in sequence table：Nucleotide shown in 1
Sequence by one or several nucleotide substitution and/or deletion and/or addition and with SEQ ID № in sequence table：Core shown in 1
Nucleotide sequence nucleotide sequence with the same function；Specifically, the function includes at least one of following (1)-(4)：
It (1) can SEQ ID № in specific recognition or binding sequence table：Nucleotide sequence shown in 3；(2) it specific recognition or can combine sequence
SEQ ID № in list：Nucleotide sequence shown in 3 by one or several nucleotide substitution and/or deletion and/or addition
Nucleotide sequence；It (3), can be with SEQ ID № in mercury, sequence table in the presence of mercury：Nucleotide sequence shown in 2 forms T-Hg
(II)-T structure；(4) in the presence of mercury, can with mercury, by SEQ ID № in sequence table：Nucleotide sequence shown in 2 by one or
The nucleotide sequence of substitution and/or the deletion and/or addition of several nucleotide forms T-Hg (II)-T structure；
2) SEQ ID № in sequence table：Nucleotide sequence shown in 2；Or by SEQ ID № in sequence table：Nucleotide shown in 2
Sequence by one or several nucleotide substitution and/or deletion and/or addition and with SEQ ID № in sequence table：Core shown in 2
Nucleotide sequence nucleotide sequence with the same function；Specifically, the function includes at least one of following (1)-(2)：
It (1), can be with SEQ ID № in mercury, sequence table in the presence of mercury：Nucleotide sequence shown in 1 forms T-Hg (II)-T structure；(2)
In the presence of mercury, can with mercury, by SEQ ID № in sequence table：Nucleotide sequence shown in 1 takes by one or several nucleotide
The nucleotide sequence of generation and/or deletion and/or addition forms T-Hg (II)-T structure；
3) SEQ ID № in sequence table：Nucleotide sequence shown in 3；Or by SEQ ID № in sequence table：Nucleotide shown in 3
Sequence by one or several nucleotide substitution and/or deletion and/or addition and with SEQ ID № in sequence table：Core shown in 3
Nucleotide sequence nucleotide sequence with the same function.Specifically, the function includes at least one of following (1)-(2)：
It (1) can SEQ ID № in specific recognition or binding sequence table：Nucleotide sequence shown in 1；(2) it specific recognition or can combine sequence
SEQ ID № in list：Nucleotide sequence shown in 1 by one or several nucleotide substitution and/or deletion and/or addition
Nucleotide sequence.
A further object of the present invention is to provide a kind of Sidestream chromatography sensor, which includes described in any of the above item
Composition.
A further object of the present invention is to provide a kind of method for detecting mercury and/or mercury ion.This method includes using upper
The composition or Sidestream chromatography sensor described in face are detected.
A further object of the present invention be to provide it is a kind of obtained from any Sidestream chromatography sensor of the present invention it is to be measured
The method of object concentration first drips sample to be tested in the sample pad area of flow measurement chromatography sensor, the inspection of fluid layer analysis sensor to be measured
After showing testing result on survey line, carries out quantitative analysis and obtain testing concentration, the method also includes：
1) detection image of the testing result of Sidestream chromatography sensor is obtained and/or shown by mobile phone；
2) gray scale for calculating and/or exporting the detection line region formation of the flow measurement chromatography sensor in the detection image is strong
Angle value and peak area S；
3) the quantitative measurement standard curve S=of the Sidestream chromatography sensor is inputted into cell phone software manually
693.71lgC-1360.4, R2=0.9868, wherein lgC is the logarithm of testing concentration, and S is peak area described in step 2)
S；
4) peak area S value obtained by step 2) is substituted into step 3) the quantitative measurement standard curve, calculates and exports to be measured
The concentration value of determinand in sample completes detection work；
Wherein, the gray-scale intensity value and described that the detection line region of the flow measurement chromatography sensor in the detection image is formed
The calculation method of peak area S value includes：
Using flow direction of the sample to be tested in the detection image on flow measurement chromatography sensor as the direction of abscissa, indulge
Coordinate is vertical with the abscissa, and the gray value Y's at all ordinates in the detection image with identical abscissa x is flat
Mean value is denoted as the gray-scale intensity value y of column, establishes gray-scale intensity P (x) function with the gray-scale intensity value y and abscissa value x of gained column
Curve；
The gray value Y calculation method is：Y=0.299R+0.587G+0.114B, wherein R, G, B be pixel R, G,
B value；
The resolution ratio is that the gray-scale intensity function curve of the detection image of m × n is：
In the gray-scale intensity function curve, Y is gray value, and x is abscissa value, and y is ordinate value, and m, n are detection figure
The resolution ratio of picture；
The peak face of the gray-scale intensity function curve in detection line region is selected, its peak area of integral calculation is to get peak face
Product S.
Of the invention a further object is provides a kind of storage medium, and the storage medium includes the program of storage, wherein
Any method of the present invention is executed by processor in described program operation.
It is of the invention that a kind of quantitative detecting analysis system that provides is provided, including：
Image capture module, image interception module, area image processing module, standard curve module, described image acquisition
For calling camera to carry out image taking or reading image from mobile phone storage device, described image interception module is used for module
The part for needing to detect in interception described image, the area image processing module are used to calculate the pixel grey scale of the part
Value constructs gray-scale intensity function, selects peak face according to the gray-scale intensity function and calculates peak face area S；The standard curve
Module is for inputting standard curve and calculating and/or export product to be tested concentration；
Wherein, the gray-scale intensity function, peak area S calculation method include：
Using flow direction of the sample to be tested in described image on flow measurement paper base chromatography sensor as the direction of abscissa, indulge
Coordinate is vertical with the abscissa, the average value of the gray value Y at all ordinates in described image with identical abscissa x
It is denoted as the gray-scale intensity value y of column, gray-scale intensity function curve is established with the gray-scale intensity value y and abscissa value x of gained column；
The gray value Y calculation method is：Y=0.299R+0.587G+0.114B, wherein R, G, B be pixel R, G,
B value；
The resolution ratio is that the gray-scale intensity function curve of the image of m × n is：
In the gray-scale intensity function curve, Y is gray value, and x is abscissa value, and y is ordinate value, and m, n are image
Resolution ratio；
Select the peak of the gray-scale intensity function curve in the detection line region in image on flow measurement paper base chromatography sensor
Face, its peak area of integral calculation is to get peak area S.
Specifically, the preparation method of the standard curve includes：
1) multiple standard items samples are provided, wherein the concentration of the determinand in multiple standard items samples is pressed identical multiple
Dilution；
2) the multiple standard items sample is detected respectively using effluent paper base chromatography sensor, is distinguished by mobile phone
Obtain and/or show the detection image of the testing result of effluent paper base chromatography sensor；
3) calculate and/or export the inspection of the flow measurement paper base chromatography sensor in the detection image of the multiple standard items sample
Multiple peak area S that survey line region is formed；
4) with the logarithm lgC of testing concentration value C or testing concentration value C in multiple standard items samples for horizontal seat
Mark, does figure using the resulting multiple multiple peak area S values corresponding from different testing concentration values of step 3) as ordinate, obtains multiple
Multiple discrete point connections are in line by discrete point, and the slope of straight line is standard curve S=a × C+b or S=a × lgC+
The intercept of slope value a in b, straight line and axis of abscissas is values of intercept b, and wherein C is testing concentration, and S is peak area S.
It is also another object of the present invention to provide any compositions of the present invention, any Sidestream chromatography sensing of invention
Device, any the method for invention, any storage medium of invention, any systematic difference of invention.
Specifically, the application includes qualitative detection or quantitative detection mercury ion
Detection method has at least the following advantages compared with other detection techniques：
(1) nucleic acid base mispairing Sidestream chromatography sensor：Using gold nano grain as signal, using can be with specific recognition mercury
Ion forms " T-Hg (II)-T " structure rich in the mercury ion in thymine alkali bases (T) nucleic acid sequence and water sample to be measured,
The cognizable red lines of naked eyes are presented in detection line, the depth of line color and the concentration of mercury ion are positively correlated, at least solve
It has determined in water the quick identification of mercury ion and the problem of its concentration is quickly converted to reliable optical signalling.
(2) smart phone Image analysis system：It is developed based on Android system, including at human-computer interaction interface and image
Adjustment method designs two parts, and to realize the rapid quantitative detection of Sidestream chromatography sensor, user can directly be read by the system
The concentration values for the target to be measured for taking Sidestream chromatography sensor to detect, solving at least traditional quantitative methods need to additionally use
The problem of volume big, expensive, irremovable instrument.
(3) Sidestream chromatography sensor provided by the invention based on nucleic acid base mispairing and/or the intelligent hand being used cooperatively
Machine Image analysis system, only produces signal response to mercury ion, and the specificity of detection is good；The lowest detection line that may be implemented is
The mercury ion of 10nM, can in the range of linearity of 10nM to 1mM in liquid mercury ion carry out quantitative detection, detection it is sensitive
Degree is high.
(4) Sidestream chromatography sensor provided by the invention based on nucleic acid base mispairing and/or the intelligent hand being used cooperatively
Machine Image analysis system is very suitable for untrained personnel and carries out on-the-spot test, existing for food safety, Environmental security etc.
Field detecting provides great convenience.
Detailed description of the invention
The drawings described herein are used to provide a further understanding of the present application, constitutes part of this application, not
Constitute the improper restriction to the application.In the accompanying drawings：
Fig. 1 is the schematic diagram of the Sidestream chromatography sensor based on nucleic acid base mispairing, wherein serial number 1-5 successively represents modeling
Expect low lining, NC film, bonding pad, water absorption pad (paper), sample pad.
Fig. 2 is the specificity experiments result figure of the Sidestream chromatography sensor based on nucleic acid base mispairing, wherein serial number 1-13
Hg (II), Zn (II), Mg (II), Pb (II), Fe (III), Fe (II), Cu (II), K (I), Ca (II), Mn are successively represented respectively
(II), the testing result of Ag (I), Au (III), Ni (II) solution.
Fig. 3 is the photo for the testing result that Sidestream chromatography sensor is shown.
Fig. 4 is optical density scatter chart.
Fig. 5 is the photo for the testing result that Sidestream chromatography sensor is shown, wherein serial number 0-9 successively represents mercury respectively
Ion concentration is feminine gender, 1nM, 10nM, 100nM, 1 μM, 10 μM, 100 μM, the testing result of 1mM, 10mM, 100mM.
Fig. 6 be optical density scatter chart, wherein serial number 0-9 successively represent respectively ion concentration of mercury as feminine gender, 1nM,
Optical density scatter chart when 10nM, 100nM, 1 μM, 10 μM, 100 μM, 1mM, 10mM, 100mM.
Fig. 7 is the corresponding relationship curve graph of ion concentration of mercury in peak area and mercury standard solution.
Fig. 8 is the canonical plotting of peak area and ion concentration of mercury.
Fig. 9 is a kind of structural schematic diagram of quantitative detecting analysis system.
Specific embodiment
Made in following embodiments, experimental method is conventional method unless otherwise specified.
Do not make the experimental methods of molecular biology illustrated, equal reference in following embodiments《Molecular Cloning:A Laboratory guide》
Listed specific method carries out in one book of (third edition) J. Pehanorm Brooker, or carries out according to kit and product description.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Following embodiments and its illustrate the improper limit not constituted for explanation and understanding the application to the application
It is fixed.
The preparation of embodiment 1, Sidestream chromatography sensor based on nucleic acid base mispairing
(1) design for the nucleotide sequence of detection
Sequence 1 (nucleotide sequence on gold nano grain)：5'-ThioMC6-GGTGGTGGTGGTGG-3'
Sequence 2 (nucleotide sequence in detection line)：5'-Biotin-CCCCCCCTCCTCCTCCTCC-3'
Sequence 3 (nucleotide sequence on nature controlling line)：5'-Biotin-CCCCCCCACCACCACCACC-3'
All nucleotide sequences of above-mentioned design pass through artificial synthesized acquisition.Wherein sequence 1 is by SEQ in sequence table
ID №：It is obtained after 5 ' end progress ThioMC6 sulfydryl modifications of nucleotide sequence shown in 1；Sequence 2 is by SEQ in sequence table
ID №：It is obtained after 5 ' end progress biotin labelings of nucleotide sequence shown in 2；Sequence 3 is by SEQ ID № in sequence table：
It is obtained after 5 ' end progress biotin labelings of nucleotide sequence shown in 3.
(2) preparation of the Sidestream chromatography sensor based on nucleic acid base mispairing
1, the nucleic acid sequence rich in thymine alkali bases for being named as sequence 2 of above-mentioned design is fixed on to the detection of NC film
On line (Test Line, T line), specific fixation procedure can refer to document Nan Cheng, Yuancong Xu, Kunlun Huang,
Yuting Chen,Zhanshen Yang,Yunbo Luo,Wentao Xu.One-step competitive lateral
flow biosensor running on an independent quantification system for smart
phones based in-situ detection of trace Hg(II)in water.Food Chemistry,2017,
214:169-175.
2, by the nucleic acid sequence and gold nano grain coupling rich in thymine alkali bases for being named as sequence 1 of above-mentioned design
Together；The preparation of gold nano grain and coupling process can refer to bibliography described in above-mentioned steps 1.
3, the nucleic acid sequence of gold nano grain is fixed on bonding pad above-mentioned coupling, specific fixation procedure can refer to
State bibliography described in step 1.
4, the nucleic acid sequence rich in adenine base for being named as sequence 3 of above-mentioned design is fixed on to the nature controlling line of NC film
On (Control Line, C line), specific fixation procedure is see also bibliography described in above-mentioned steps 1.
5, the above-mentioned NC film prepared and bonding pad are prepared into Sidestream chromatography sensor according to existing method, specifically may be used
Choosing：As shown in Figure 1, the NC film prepared to be fixed to the middle part of the low lining 1 of plastics；The bonding pad prepared is covered into NC film 2
One end so that bonding pad 3 and NC film 2 partly overlaps；Water absorption pad (paper) 4 is covered to the other end of NC film 2, so that NC film 2
It partly overlaps with water absorption pad (paper) 4；Sample pad 5 is covered into the one end of bonding pad 3 far from NC film 2, and makes bonding pad 3 and sample
Product pad 5 partly overlaps；Last covering protection film, it is spare to be prepared into Sidestream chromatography sensor.
The NC film 2, bonding pad 3, sample pad 5, material is followed successively by nitrocellulose respectively used in water absorption pad (paper) 4
Film, glass fibre membrane, glass fibre membrane, blotting paper.
(3) testing principle and process of the Sidestream chromatography sensor based on nucleic acid base mispairing
The testing principle of nucleic acid base mispairing Sidestream chromatography sensor is based on sandwich structure (rich in thymine alkali bases
Nucleic acid sequence-mercury ion-is rich in thymine alkali bases nucleic acid sequence), as shown in Figure 1, being rich in thymine alkali bases core for one section
Acid sequence is fixed in detection line, and another section is coupled together admittedly rich in thymine alkali bases nucleic acid sequence and gold nano grain
It is scheduled on bonding pad, one section is fixed on nature controlling line rich in adenine base nucleic acid sequence.In the detection of a standard, contain
There is the sample of a certain concentration mercury ion to be dripped first in sample pad, solution is due to capillary force (i.e. water absorption pad or paper later
Suction) arrival bonding pad can be moved up with the direction of chromatography sensor；With on bonding pad be rich in thymine alkali bases nucleic acid
Sequence continues to move up arrival detection line with the direction of chromatography sensor together with the compound that gold nano grain is coupled；?
In detection line, a certain concentration mercury ion is rich in conjunction with thymine alkali bases nucleic acid sequence with two sections in sample, forms " T-Hg
(II)-T " structure makes gold nano grain be crawled and be piled up in detection line, occurs the cognizable red line of naked eyes in detection line
Item, ion concentration of mercury is bigger in sample, red deeper；It is excessive even rich in thymine alkali bases nucleic acid sequence and gold nano grain
The compound of connection continues to move up arrival nature controlling line, is combined by the base pair complementarity of thymidine and adenine, makes gold
Nano particle is crawled and is piled up in nature controlling line, occurs the cognizable red lines of naked eyes on nature controlling line.If not contained in sample
A certain concentration mercury ion, then can not form " T-Hg (II)-T " structure in detection line, the also not accumulation of gold nano grain, because
It is not in the cognizable red lines of naked eyes in this detection line.
The specificity experiments of embodiment 2, Sidestream chromatography sensor based on nucleic acid base mispairing
Using embodiment 1 it is preparation-obtained based on the Sidestream chromatography sensor of nucleic acid base mispairing to different metals from
Sub- solution is detected, and the specificity of the sensor is tested, wherein Hg (II) concentration is that 1 μM of other concentration of metal ions is
1mM。
Sample to be tested is dripped in sample pad, about after five minutes, Sidestream chromatography sensor can show testing result.Specifically
Property experimental result as shown in Fig. 2, should Sidestream chromatography sensor based on nucleic acid base mispairing signal only was produced to mercury ion rang
It answers, illustrates that this method specificity is good.
Embodiment 3, the quantitative detection that Sidestream chromatography sensor is realized by smart phone Image analysis system
Mercury ion is detected according to disclosure specific embodiment, that is, Sidestream chromatography described in embodiment 1 sensor
When, red lines can be showed in detection line and/or nature controlling line.Therefore the image comprising red lines is utilized, by hand
Establish corresponding with ion concentration of mercury standard curve in machine, i.e., image using shooting comprising red lines (or in advance
First in mobile phone storage, or the image for passing through mobile phone-downloaded) quantitative detection is carried out to ion concentration of mercury.
(1) foundation of standard curve
Prepare a series of mercury standard solutions by the diluted known concentration of multiple, ion concentration of mercury in different mercury standard solutions
It is followed successively by 0,1nM, 10nM, 100nM, 1 μM, 10 μM, 100 μM, 1mM, 10mM, 100mM.
The mercury standard solution of above-mentioned prepared various concentration is dripped prepared by 10 embodiments 1 respectively based on nucleic acid
In the sample pad of the Sidestream chromatography sensor of base mispairing, about after five minutes, Sidestream chromatography sensor can show testing result.
It is taken pictures, be can be obtained by the testing result that the camera of mobile phone respectively shows above-mentioned Sidestream chromatography sensor
As shown in Figure 3 is presented the image of red lines in nature controlling line and/or actual measurement line, which is also possible to be stored in advance in mobile phone
On image, or by mobile phone-downloaded obtain image.
Then, using the longitudinal extension of Sidestream chromatography sensor as abscissa, with the flat of the corresponding column of each abscissa
Equal gray value is ordinate, and curve image as shown in Figure 4 (i.e. optical density distribution curve) can be obtained.
The curve of detection line position is chosen, the peak area for calculating curve at this (carries out integral fortune to the curve
Calculate), peak area value corresponding with specific ion concentration of mercury can be obtained.
In the above manner, being taken pictures according to as shown in Figure 5 as a result, obtaining optical density distribution curve as shown in FIG. 6, root
According to curve obtained, calculate separately out different ion concentration of mercury values (for example, 0,1nM, 10nM, 100nM, 1 μM, 10 μM, 100 μM,
1mM, 10mM, 100mM) corresponding peak area value.As shown in fig. 7, using obtained peak area to known mercury standard solution
The concentration value of middle mercury ion makes curve.Standard curve as shown in Figure 8 is calculated finally by Excel artificial fitting：S=
693.71lgC-1360.4, R2=0.9868, wherein lgC is the logarithm of determinand ion concentration of mercury, and S represents peak area value.
(2) foundation of smart phone Image analysis system
Above-mentioned the Fitting Calculation is obtained into standard curve：S=693.71lgC-1360.4 (R2=0.9868) it is built into intelligence
In mobile phone Image analysis system；
Sample to be tested is dripped in sample pad, about after five minutes, Sidestream chromatography sensor can show testing result, pass through
The camera of mobile phone takes pictures to the testing result, obtains detection image, and result of taking pictures can be stored into mobile phone, also can be used directly；
The calculation method of gray-scale intensity value and peak area S value in detection image is：
Using flow direction of the sample to be tested in the detection image on flow measurement chromatography sensor as the direction of abscissa, indulge
Coordinate is vertical with the abscissa, and the gray value Y's at all ordinates in the detection image with identical abscissa x is flat
Mean value is denoted as the gray-scale intensity value y of column, establishes gray-scale intensity P (x) function with the gray-scale intensity value y and abscissa value x of gained column
Curve；
The gray value Y calculation method is：Y=0.299R+0.587G+0.114B, wherein R, G, B be pixel R, G,
B value；
The resolution ratio is that the gray-scale intensity function curve of the detection image of m × n is：
In the gray-scale intensity function curve, Y is gray value, and x is abscissa value, and y is ordinate value, and m, n are detection figure
The resolution ratio of picture；
The peak face of the gray-scale intensity function curve in detection line region is selected, its peak area of integral calculation is to get peak face
Product S.
By gained peak area S input built-in standard curve, calculate output concentration value C, that is, output captured inspection
Survey concentration value corresponding to the detection line in result photo.
In a specific embodiment, the present embodiment smart phone Image analysis system can be carried out based on Android system
Exploitation, design.
(3) mercury ion of the Sidestream chromatography sensor based on nucleic acid base mispairing and smart phone Image analysis system is fixed
Measure the sensitivity of detection
As shown in Figure 7, Figure 8, using peak area obtained by smart phone Image analysis system described in the present embodiment and mercury from
Sub- concentration has good correlation；Sidestream chromatography sensor based on nucleic acid base mispairing and this implementation prepared by embodiment 1
The mercury ion that the lowest detection line that smart phone Image analysis system described in example may be implemented is 10nM, high sensitivity；Linearly
Range is 10nM to 1mM, can carry out quantitative detection to mercury ion in water in this range.
In addition, Fig. 9 shows a kind of structural schematic diagram of quantitative detecting analysis system, including：
Image capture module, image interception module, area image processing module, standard curve module, described image acquisition
For calling camera to carry out image taking or reading image from mobile phone storage device, described image interception module is used for module
The part for needing to detect in interception described image, the area image processing module are used to calculate the pixel grey scale of the part
Value constructs gray-scale intensity function, selects peak face according to the gray-scale intensity function and calculates peak face area S；The standard curve
Module is for inputting standard curve and calculating and/or export product to be tested concentration；
Wherein, the gray-scale intensity function, peak area S calculation method include：
Using flow direction of the sample to be tested in described image on flow measurement paper base chromatography sensor as the direction of abscissa, indulge
Coordinate is vertical with the abscissa, the average value of the gray value Y at all ordinates in described image with identical abscissa x
It is denoted as the gray-scale intensity value y of column, gray-scale intensity function curve is established with the gray-scale intensity value y and abscissa value x of gained column；
The gray value Y calculation method is：Y=0.299R+0.587G+0.114B, wherein R, G, B be pixel R, G,
B value；
The resolution ratio is that the gray-scale intensity function curve of the image of m × n is：
In the gray-scale intensity function curve, Y is gray value, and x is abscissa value, and y is ordinate value, and m, n are image
Resolution ratio；
Select the peak of the gray-scale intensity function curve in the detection line region in image on flow measurement paper base chromatography sensor
Face, its peak area of integral calculation is to get peak area S.
The preparation method of the standard curve includes：
1) multiple standard items samples are provided, wherein the concentration of the determinand in multiple standard items samples is pressed identical multiple
Dilution；
2) the multiple standard items sample is detected respectively using effluent paper base chromatography sensor, is distinguished by mobile phone
Obtain and/or show the detection image of the testing result of effluent paper base chromatography sensor；
3) calculate and/or export the inspection of the flow measurement paper base chromatography sensor in the detection image of the multiple standard items sample
Multiple peak area S that survey line region is formed；
4) with the logarithm lgC of testing concentration value C or testing concentration value C in multiple standard items samples for horizontal seat
Mark, does figure using the resulting multiple multiple peak area S values corresponding from different testing concentration values of step 3) as ordinate, obtains multiple
Multiple discrete point connections are in line by discrete point, and the slope of straight line is standard curve S=a × C+b or S=a × lgC+
The intercept of slope value a in b, straight line and axis of abscissas is values of intercept b, and wherein C is testing concentration, and S is peak area S.
Claims (10)
- At least one of 1. a kind of composition, which is characterized in that the composition includes following 1) -3)：1) SEQ ID № in sequence table：Nucleotide sequence shown in 1；Or by SEQ ID № in sequence table：Nucleotide sequence shown in 1 By one or several nucleotide substitution and/or deletion and/or addition and with SEQ ID № in sequence table：Nucleotide shown in 1 Sequence nucleotide sequence with the same function；2) SEQ ID № in sequence table：Nucleotide sequence shown in 2；Or by SEQ ID № in sequence table：Nucleotide sequence shown in 2 By one or several nucleotide substitution and/or deletion and/or addition and with SEQ ID № in sequence table：Nucleotide shown in 2 Sequence nucleotide sequence with the same function；3) SEQ ID № in sequence table：Nucleotide sequence shown in 3；Or by SEQ ID № in sequence table：Nucleotide sequence shown in 3 By one or several nucleotide substitution and/or deletion and/or addition and with SEQ ID № in sequence table：Nucleotide shown in 3 Sequence nucleotide sequence with the same function.
- 2. composition according to claim 1, which is characterized in that the composition further includes following 1) -3) at least It is a kind of：1) SEQ ID № in the sequence table：The 5 ' of nucleotide sequence shown in 1 are terminal modified sulfydryl；2) SEQ ID № in the sequence table：5 ' ends of nucleotide sequence shown in 2 are marked with biotin；3) SEQ ID № in the sequence table：5 ' ends of nucleotide sequence shown in 3 are marked with biotin.
- 3. a kind of Sidestream chromatography sensor, which is characterized in that the Sidestream chromatography sensor includes that claim 1 and/or 2 is any The composition.
- 4. Sidestream chromatography sensor according to claim 3, the sensor include nature controlling line and detection line, feature exists In, the Sidestream chromatography sensor further include following 1) -4) at least one of：1) SEQ ID № in the sequence table：Nucleotide sequence shown in 2 is located in detection line；2) the Sidestream chromatography sensor further includes bonding pad, SEQ ID № in the sequence table：Nucleotide sequence shown in 1 with Gold nano grain is coupled together and is located on bonding pad；3) SEQ ID № in the sequence table：Nucleotide sequence shown in 3 is located on nature controlling line；4) the Sidestream chromatography sensor further includes low lining, NC film, bonding pad, water absorption pad or paper, sample pad；The NC film is located at The middle part of low lining；One end of the bonding pad and NC film partly overlaps, the other end part weight of the water absorption pad or paper and NC film It is folded；The sample pad and bonding pad partly overlap far from one end of NC film；The nature controlling line and detection line are respectively positioned on the NC film On, wherein nature controlling line is located at one end close to water absorption pad or paper, and detection line is located at close to one end of bonding pad.
- 5. a kind of method of detection mercury and/or mercury ion, which is characterized in that the method includes using claim 1 and/or 2 The composition, or detected using claim 3 and/or the 4 Sidestream chromatography sensors.
- 6. a kind of method for obtaining testing concentration from claim 3 and/or the 4 Sidestream chromatography sensors, first will be to be measured Sample drop chromatographs the sample pad area of sensor in flow measurement, after showing testing result in the detection line of fluid layer analysis sensor to be measured, It carries out quantitative analysis and obtains testing concentration, which is characterized in that the method also includes：1) detection image of the testing result of Sidestream chromatography sensor is obtained and/or shown by mobile phone；2) calculate and/or export the gray-scale intensity value that the detection line region of the flow measurement chromatography sensor in the detection image is formed With peak area S；3) the quantitative measurement standard curve S=693.71lgC- of the Sidestream chromatography sensor is inputted into cell phone software manually 1360.4 R2=0.9868, wherein lgC is the logarithm of testing concentration, and S is peak area S described in step 2)；4) peak area S value obtained by step 2) is substituted into step 3) the quantitative measurement standard curve, calculates and exports sample to be tested The concentration value of middle determinand completes detection work；Wherein, the gray-scale intensity value and the peak face that the detection line region of the flow measurement chromatography sensor in the detection image is formed Product S value calculation method include：Using flow direction of the sample to be tested in the detection image on flow measurement chromatography sensor as the direction of abscissa, ordinate It is vertical with the abscissa, the average value of the gray value Y at all ordinates in the detection image with identical abscissa x It is denoted as the gray-scale intensity value y of column, gray-scale intensity P (x) function curve is established with the gray-scale intensity value y and abscissa value x of gained column；The gray value Y calculation method is：Y=0.299R+0.587G+0.114B, wherein R, G, B are R, G, B value of pixel；The resolution ratio is that the gray-scale intensity function curve of the detection image of m × n is：In the gray-scale intensity function curve, Y is gray value, and x is abscissa value, and y is ordinate value, and m, n are detection image Resolution ratio；The peak face of the gray-scale intensity function curve in detection line region is selected, its peak area of integral calculation is to get peak area S.
- 7. a kind of storage medium, the storage medium includes the program of storage, wherein is held in described program operation by processor Row method of claim 6.
- 8. a kind of quantitative detecting analysis system, which is characterized in that including：Image capture module, image interception module, area image processing module, standard curve module, described image acquisition module For calling camera to carry out image taking or reading image from mobile phone storage device, described image interception module is for intercepting The part for needing to detect in described image, the area image processing module are used to calculate the grey scale pixel value of the part, structure Gray-scale intensity function is built, peak face is selected according to the gray-scale intensity function and calculates peak face area S；The standard curve module is used In input standard curve and calculating and/or output product to be tested concentration；Wherein, the gray-scale intensity function, peak area S calculation method include：Using flow direction of the sample to be tested in described image on flow measurement paper base chromatography sensor as the direction of abscissa, ordinate Vertical with the abscissa, the average value of the gray value Y at all ordinates in described image with identical abscissa x is denoted as The gray-scale intensity value y of column establishes gray-scale intensity function curve with the gray-scale intensity value y and abscissa value x of gained column；The gray value Y calculation method is：Y=0.299R+0.587G+0.114B, wherein R, G, B are R, G, B value of pixel；The resolution ratio is that the gray-scale intensity function curve of the image of m × n is：In the gray-scale intensity function curve, Y is gray value, and x is abscissa value, and y is ordinate value, and m, n are the resolution of image Rate；Select the peak face of the gray-scale intensity function curve in the detection line region in image on flow measurement paper base chromatography sensor, product Divide and calculates its peak area to get peak area S.
- 9. system according to claim 8, which is characterized in that the preparation method of the standard curve includes：1) multiple standard items samples are provided, wherein the concentration of the determinand in multiple standard items samples is dilute by identical multiple It releases；2) the multiple standard items sample is detected respectively using effluent paper base chromatography sensor, is obtained respectively by mobile phone And/or the detection image of the testing result of display effluent paper base chromatography sensor；3) calculate and/or export the detection line of the flow measurement paper base chromatography sensor in the detection image of the multiple standard items sample Multiple peak area S that region is formed；4) using the logarithm lgC of testing concentration value C or testing concentration value C in multiple standard items samples as abscissa, with The resulting multiple multiple peak area S values corresponding from different testing concentration values of step 3) are that ordinate does figure, are obtained multiple discrete Point, multiple discrete points connections are in line, the slope of straight line is in standard curve S=a × C+b or S=a × lgC+b Slope value a, the intercept of straight line and axis of abscissas is values of intercept b, and wherein C is testing concentration, and S is peak area S.
- 10. claim 1 and/or 2 compositions, claim 3 and/or the 4 Sidestream chromatography sensors, claim 5 Or storage medium, claim 8 and/or 9 systematic differences described in 6 the methods, claim 7.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810688361.8A CN108844951B (en) | 2018-06-28 | 2018-06-28 | Mercury ion detection product and method and smartphone imaging analysis system |
| PCT/CN2018/100985 WO2020000598A1 (en) | 2018-06-28 | 2018-08-17 | Mercury ion detection product and method and smart phone imaging analysis system |
| CA3053677A CA3053677A1 (en) | 2018-06-28 | 2018-08-17 | Product, method and smartphone imaging analysis system for mercury ion detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810688361.8A CN108844951B (en) | 2018-06-28 | 2018-06-28 | Mercury ion detection product and method and smartphone imaging analysis system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108844951A true CN108844951A (en) | 2018-11-20 |
| CN108844951B CN108844951B (en) | 2021-07-13 |
Family
ID=64200585
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810688361.8A Active CN108844951B (en) | 2018-06-28 | 2018-06-28 | Mercury ion detection product and method and smartphone imaging analysis system |
Country Status (3)
| Country | Link |
|---|---|
| CN (1) | CN108844951B (en) |
| CA (1) | CA3053677A1 (en) |
| WO (1) | WO2020000598A1 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10991185B1 (en) | 2020-07-20 | 2021-04-27 | Abbott Laboratories | Digital pass verification systems and methods |
| CN112285078B (en) * | 2020-10-14 | 2023-04-25 | 安庆师范大学 | Novel method for detecting mercury ions on site based on intelligent DNA hydrogel |
| CN113720837B (en) * | 2021-09-23 | 2024-01-19 | 西北大学 | Colorimetric sensor for rapidly detecting mercury ions in water body |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608108A (en) * | 2012-02-21 | 2012-07-25 | 温州医学院 | Nano biosensor method and kit for rapid detection of Hg ions |
| CN105886618A (en) * | 2016-04-21 | 2016-08-24 | 中国农业大学 | Method for quantitatively detecting mercury ions in liquid sample and kit |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1657919A (en) * | 2005-03-11 | 2005-08-24 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Mercury investigating test paper and its preparation method |
| CN1687752A (en) * | 2005-04-29 | 2005-10-26 | 农业部环境保护科研监测所 | Test paper for detecting heavy metal mercury rapidly, preparation method and application |
| CN104076004A (en) * | 2014-07-16 | 2014-10-01 | 常熟理工学院 | Method for detecting mercury ionic concentration in sample |
| CN104931491B (en) * | 2015-06-02 | 2018-03-27 | 北京福德安科技有限公司 | A kind of 61 test paper of conjunction for heavy-metal residual quick detection |
| CN205003082U (en) * | 2015-09-06 | 2016-01-27 | 金陵科技学院 | Detect aptamer test paper of mercury ion |
-
2018
- 2018-06-28 CN CN201810688361.8A patent/CN108844951B/en active Active
- 2018-08-17 CA CA3053677A patent/CA3053677A1/en active Pending
- 2018-08-17 WO PCT/CN2018/100985 patent/WO2020000598A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608108A (en) * | 2012-02-21 | 2012-07-25 | 温州医学院 | Nano biosensor method and kit for rapid detection of Hg ions |
| CN105886618A (en) * | 2016-04-21 | 2016-08-24 | 中国农业大学 | Method for quantitatively detecting mercury ions in liquid sample and kit |
Non-Patent Citations (3)
| Title |
|---|
| CHENG，ET.AL.: ""One-step competitive lateral flow biosensor running on an independent quantification system for smart phones based in-situ detection of trace Hg(II) in tap water"", 《FOOD CHEMISTRY》 * |
| 何雨希: ""智能手机用于检测水体中几种典型无机污染物的研究"", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 李好枝: "《体内药物分析》", 31 August 2003, 中国医药科技出版社 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108844951B (en) | 2021-07-13 |
| CA3053677A1 (en) | 2019-12-28 |
| WO2020000598A1 (en) | 2020-01-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108844951A (en) | A kind of mercury ion detecting product, method and smart phone Image analysis system | |
| CN105861299B (en) | Droplet type digital pcr fluorescence detecting system and fluorescence detection device | |
| Hanna | Testing for HER2 status | |
| Bowie et al. | Real-time monitoring of picomolar concentrations of iron (II) in marine waters using automated flow injection-chemiluminescence instrumentation | |
| Kitidis et al. | Seasonal dynamics of the carbonate system in the Western English Channel | |
| CN104297468B (en) | The calibration steps of a kind of immune fetch equipment and this equipment | |
| CN101210922B (en) | Method for detecting SPA FC fragment bound mammalian blood serum total IgG by dolloidal gold marker protein A | |
| Nežerka et al. | Assessment of aggregate-bitumen coverage using entropy-based image segmentation | |
| Newlander et al. | Integrating visual and chemical data to source chert artifacts in the North American Great Basin | |
| CN106485250A (en) | A kind of recognition methods of paper chip gray value and its application | |
| CN104931488B (en) | A method of based on indicator array detection anion | |
| US20130130243A1 (en) | Method and device for detecting and quantifying an analyte with recycling of the reagents | |
| CN107917876B (en) | Antibiotic detection device and method based on nanogold-aptamer structure | |
| CA2815205C (en) | A method of measuring a capacitance and a use | |
| CN108333085B (en) | Photoelectric vertical line sand content detection method with capacitance and color compensation | |
| CN103063674A (en) | Detection method for copper grade of copper block, and detection system thereof | |
| Qi et al. | Dual-Mode Undistorted Visual Fluorescent Sensing Strategy through Manipulating the Coffee-Ring Effect on Microfluidic Paper-Based Chip | |
| CN103713035A (en) | Electrochemical detection method of okadaic acid in shellfish | |
| CN110006857A (en) | A kind of iodide ion quantifies instant detection technique | |
| CN108603883A (en) | The determination method of prostate cancer | |
| CN109060802B (en) | Paper-based chromatographic sensor quantitative analysis system and analysis method based on mobile phone | |
| CN101639444B (en) | Method for nano particle reinforced fluorescence polarization analysis | |
| CN103616522A (en) | Immune chromatography result identification method based on envelope area and twice compensation | |
| Zhao et al. | Application of the Ludox-QPS method for estimating ciliate diversity in soil and comparison with direct count and DNA fingerprinting | |
| Holmquist et al. | Luminescence immunoassay of pollen allergens on air sampling polytetrafluoroethylene filters |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Xu Wentao Inventor after: Luo Yunbo Inventor after: Cheng Nan Inventor after: Huang Kunlun Inventor after: Xu Aicong Inventor after: Yang Zhansen Inventor before: Luo Yunbo Inventor before: Xu Wentao Inventor before: Cheng Nan Inventor before: Huang Kunlun Inventor before: Xu Yuancong Inventor before: Yang Zhansen |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |