CN106198418A - A kind of photometric detection method and system - Google Patents
A kind of photometric detection method and system Download PDFInfo
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- CN106198418A CN106198418A CN201610595574.7A CN201610595574A CN106198418A CN 106198418 A CN106198418 A CN 106198418A CN 201610595574 A CN201610595574 A CN 201610595574A CN 106198418 A CN106198418 A CN 106198418A
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- G—PHYSICS
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- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
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- 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
Abstract
A kind of open photometric detection method of the present invention and detecting system, this detection method includes step: smart mobile phone scans testing sample and the label of standard sample；Smart mobile phone is according to scanning the testing sample and the kind of standard sample obtained, the photometric detection operational approach that display is corresponding；Smart mobile phone carries out image acquisition to the testing sample after processing according to photometric detection operational approach and standard sample；Being scaled spectral wavelength and light intensity according to the image collected, by analyzing, calculating and process, display testing result also generates examining report.Smart mobile phone and multi-field photometric detection can be tested the application that combines by the present invention.
Description
Technical field
The present invention relates to scientific instrument detection field, in particular, relate to a kind of photometric detection method and system.
Background technology
The fast development of smart mobile phone technology, for scientific instrument design, make and use and open new field.Intelligence
The clear analysis degree of the photographic head of mobile phone and sensitivity are more and more higher, and detection spectral region is increasingly wider, and this makes can be by change machine
Carry photographic head, detection ultraviolet spectra and infrared spectrum.
Further, smart mobile phone can be incorporated into the detection in the fields such as biomedicine, chemistry, environment, food, metallurgy
In the middle of experiment, the most preferably smart mobile phone and test experience are combined, become engineers problem demanding prompt solution.
Summary of the invention
The technical problem to be solved be to provide a kind of can be real by smart mobile phone and multi-field photometric detection
Test the photometric detection method and system combined.
It is an object of the invention to be achieved through the following technical solutions:
A kind of photometric detection method, including step:
Smart mobile phone scanning testing sample and the label of standard sample；
Smart mobile phone is according to scanning the testing sample and the kind of standard sample obtained, the photometric detection operation side that display is corresponding
Method:
Smart mobile phone carries out image acquisition to the testing sample after processing according to photometric detection operational approach and standard sample；
Being scaled spectral wavelength and light intensity according to the image collected, by analyzing, calculating and process, display testing result is also
Generate examining report.
Preferably, further comprise the steps of:
Form the photometric detection behaviour at least including detected sample information, standard sample information, sample type information, detection equipment
Make the cloud data base of method, sample detection methods, detectable information, operator's information；
And upload storage detect successfully record, failure logging and operational error record, detect time cumulation, detect sample, make
Used disposable device and operator, accordingly, provide suggestion for operation.Use cloud data base that sample photometric detection is tested phase
Pass information carries out the renewal storage that prestores and follow up, and uses big data technique and cloud storage technology, analyzes and processes detection process
In details so that detection analyze convenient, be easier to successfully；And it is possible to test based on forefathers and previous photometric detection,
Follow-up luminosity test experience is instructed and advises, helps success and the progress of subsequent experimental.
Preferably, described photometric detection operational approach at least includes two steps, uses smart mobile phone to each step
The detection equipment and the reagent that use carry out record；
Smart mobile phone, according to pre-setting, carries out image acquisition, and the figure that will collect to testing sample and standard sample automatically
The rgb value conversion of picture obtains spectral wavelength and light intensity.Detection equipment and reagent that different sample types use are not necessarily the same, right
It carries out record, subsequent experimental can be carried out directive function；Even and if same sample type, its detection equipment used and examination
Agent, the most just the same, by relative analysis, the success more preferably faster of follow-up luminosity test experience can be helped and enter
Step；
It addition, have in the experiment of some photometric detection, the detection of the luminous intensity of testing result and absorbance etc. is had one
Excellent detection is interval, automatically gathers spectral wavelength, light intensity map picture, experimenter can be helped to complete luminosity the most more accurately
Test experience.
Preferably, if detected sample is pathogenic microorganism to be detected, standard sample is standard pathogenic microorganism；
Then according to corresponding photometric detection operational approach, pathogenic microorganism to be detected and standard pathogenic microorganism are carried out isothermal expansion
Increase detection；
To carrying out pathogenic microorganism to be detected and the standard pathogenic microorganism of isothermal duplication detection, carry out spectral wavelength, light intensity
Gather；
Obtain the luminosity information of pathogenic microorganism to be detected and standard pathogenic microorganism, and generate curve to be detected and the mark of correspondence
Directrix curve；
According to curve to be detected and the contrast of standard curve and analysis, obtain pathogenic microorganism to be detected and standard pathogenic microorganism
Photometric detection result；Certainly, this pathogenic microorganism to be detected can be one can also be multiple；Augmentation detection includes glimmering
Light detection by quantitative and the Turbidity measurement of magnesium pyrophosphate by-product, in fluorescent quantitation detection, utilize SYBR Green I fluorescent dye
Only with double-stranded DNA minor groove binding, when it is with DNA double chain combination, send the principle of the fluorescence of the strongest 800～1000 times,
In an individual system, its signal intensity represents the quantity of double chain DNA molecule.When nucleic acid synthesizes in a large number, SYBR Green I
Double-stranded DNA be can automatically mix, by the produced fluorescence intensity of detection, Ct value, reference standards curve, it may be determined that DNA's obtained
Starting copy number；
And in Turbidity measurement, when nucleic acid synthesizes in a large number, the pyrophosphate ion separated out from dNTP and the Mg2+ reaction solution
In conjunction with, produce magnesium pyrophosphate byproduct precipitate.Reaction equation is as follows:
(DNA)n-1 +dNTP=(DNA)n+P2O7 4-；P2O7 4-+2Mg2+=Mg2P2O7(precipitation)；
This reaction has high specificity, under 400nm light, by photographic head detection precipitation turbidity just can interpolate that amplification with
No.
Preferably, in described isothermal duplication detection, it is micro-to cause of disease to be detected that smart mobile phone controls micro-fluidic LAMP chip
Biological and standard pathogenic microorganism carries out detection operation.Use smart mobile phone photographic head as detector, any class can be identified
The multi-channel chip of type, it is achieved effective differentiation of multiple LAMP signal, portable, low in energy consumption, simple to operate, beneficially LAMP method
Promotion and application；Detection processes full-automation with data, and simple and convenient, layman also can Successful Operation；Use intelligence
Handset image processing module as LAMP chip detector, can feature pathogen in automatization's Quantitative detection sample, tool
Have highly sensitive, detection speed is fast, high specificity, simple to operate, result is stable, automatically provide measurement result, the advantage such as portable.
Preferably, described micro-fluidic LAMP chip is arranged on the chip carrier controlled by described smart mobile phone, described chip
Seat is provided with the laser assembly for providing experimental optical source, and replaceable LAMP chip interface；
Amplification pond it is provided with in described micro-fluidic LAMP chip；Amplification is provided with for detecting pathogenic microorganism to be measured and mark in pond
The specific probe of quasi-microorganism.The replaceable LAMP chip interface that chip carrier uses, and interface has extensibility, Ke Yigen
The chip of varying number sense channel is changed, it is achieved the most multiple LAMP effectively expands and multiple LAMP detection according to actual demand
Effective differentiation of signal, provides a kind of effective means for clinical multiple pathogens quick diagnosis；Use smart mobile phone as photoelectricity
Colorimetric detector, smart mobile phone does not contacts with chip under test, thus avoids sample and chemical reagent to intelligence hands
The corrosion of machine, tester, also without the electric unit of contact chip, improves safety.
Preferably, if testing sample is test serum sample, standard sample is standard serum samples；
Then according to corresponding photometric detection operational approach, blood serum sample to be measured and standard serum samples are carried out serum glycated white egg
White detection；
Test serum sample after carrying out serum glycated albumin detection and standard sample are carried out image acquisition, obtains correspondence
Spectral wavelength and light intensity；
Obtain test serum sample and standard sample absorbance variable quantity under the light source of 570-650nm wavelength irradiates, and generate
Corresponding curve to be measured and standard curve；
According to curve to be measured and the contrast of standard curve and analysis, obtain the glycosylated albumin content of test serum sample.At this
In photometric detection test, the absorbance of sample is directly proportional to the glycosylated albumin content of sample, by contrasting serum sample to be detected
Product and the absorbance curve of standard sample, can calculate the glycosylated albumin content of detected sample.
Preferably, if testing sample is air sample to be measured, standard sample is normal air sample；
Then according to corresponding photometric detection operational approach, air sample to be measured and normal air sample are carried out PM2.5 particulate matter
Concentration Testing；
Air sample to be measured after carrying out the Concentration Testing of PM2.5 particulate matter and normal air sample are carried out image acquisition,
To corresponding spectral wavelength and light intensity；
Obtain air sample to be measured and the absorbance of normal air sample and generate curve to be measured and the standard curve of correspondence；
According to curve to be measured and the contrast of standard curve and analysis, obtain the concentration of the PM2.5 particulate matter of air sample to be measured.Sample
The concentration of the PM2.5 in product, the absorbance carrying out the product after photometric detection experiment to sample is relevant；So, in photometric detection
After experiment, gather air sample to be detected and the absorbance of normal air sample by smart mobile phone, and analyze its corresponding generation
Curve to be measured and standard curve, the PM2.5 particle concentration of air sample to be detected can be calculated.
Preferably, if testing sample is solution to be measured, standard sample is standard solution；
Then according to corresponding photometric detection operational approach, solution to be measured and standard solution are carried out content of chromium ion detection；
The solution to be measured and standard solution carrying out content of chromium ion detection is carried out image acquisition, obtain correspondence spectral wavelength and
Light intensity；
Obtain solution to be measured and the standard solution absorbance under the light source of 540nm wavelength irradiates and generate the curve to be measured of correspondence
And standard curve；
According to curve to be measured and the contrast of standard curve and analysis, obtain the content of chromium ion in solution to be detected.According to intelligence
Detection operation method presentation on energy mobile phone carries out photometric detection, and after standing the scheduled time, smart mobile phone photographic head automatically turns on,
The absorbance at system display 540nm place, generates curve to be measured and the standard curve of correspondence, can automatically calculate in sample chromium from
The content of son.
Preferably, if detected sample is oils and fats to be detected, standard sample is standard oils and fats；
Then according to corresponding photometric detection operational approach, oils and fats to be detected and standard oils and fats are carried out Oxidation of Fat and Oils degree and examines
Survey；
Oils and fats to be detected and standard oils and fats are carried out image acquisition, obtains colourity and the intensity level of correspondence；
According to the oils and fats to be detected collected and the colourity of standard oils and fats and intensity level, display testing result also generates detection report
Accuse.Scanning standard solution and the label of solution to be detected, by operation indicating, scan each reagent label, and add reagent, and by referring to
Determine step operation；After colour developing, smart mobile phone shoots each test tube at the middle and upper levels or lower floor's color, according to the colourity identified with strong
Angle value, generates curve to be measured and standard curve automatically according to sample label and testing result, and draws third in solution to be detected
The concentration value of dialdehyde, thus judge the degree of oxidation of oils and fats in this sample.
The present invention, in the photometric analysis and test experience in the field such as biomedicine, chemistry, environment, food, metallurgy, introduces
The use of smart mobile phone；First, by smart mobile phone scanning detected sample and the label of standard sample, can quickly determine
The kind of sample, and show the photometric detection operational approach of correspondence, the effect of test experience is improved not only by smart mobile phone
Rate, furthermore, it is possible to make this smart mobile phone be applied in the middle of the photometric detection experiment of multiple fields；Further, due to many
In photometric detection experiment, after the process of its testing result and photometric detection, the luminous intensity of sample and absorbance etc. have and greatly contact,
And development in science and technology is so far, the information such as the luminous intensity of many of which and absorbance can be treated by the camera collection of smart mobile phone
The image of test sample product and standard sample obtains spectral wavelength, light intensity, so, utilizes smart mobile phone to may also help in experiment people
Member analyzes the result of photometric detection experiment, and corresponding generation examining report, offers convenience to experimenter.
Accompanying drawing explanation
Fig. 1 is the flow chart of the embodiment of the present invention a kind of photometric detection method.
Detailed description of the invention
The invention will be further described with preferred embodiment below in conjunction with the accompanying drawings.
Embodiment one:
Fig. 1 is the flow chart of the embodiment of the present invention a kind of photometric detection method, including step:
S1: smart mobile phone scanning testing sample and the label of standard sample；
S2: smart mobile phone is according to scanning the testing sample and the kind of standard sample obtained, and the photometric detection of display correspondence operates
Method:
S3: smart mobile phone carries out image acquisition to the testing sample after processing according to photometric detection operational approach and standard sample；
S4: be scaled spectral wavelength and light intensity according to the image collected, is then analyzed, calculates and processes, display inspection
Survey result and generate examining report.
The present invention, in the photometric analysis and test experience in the field such as biomedicine, chemistry, environment, food, metallurgy, introduces
The use of smart mobile phone；First, by smart mobile phone scanning detected sample and the label of standard sample, sample can quickly be determined
The kind of product, and show the photometric detection operational approach of correspondence, the efficiency of test experience is improved not only by smart mobile phone,
Furthermore, it is possible to make this smart mobile phone be applied in the middle of the photometric detection experiment of multiple fields；
Further, in testing due to many photometric detection, the luminous intensity of sample after its testing result and photometric detection process
Having greatly contact with absorbance etc., and development in science and technology is so far, the information such as the luminous intensity of many of which and absorbance can be passed through
The camera collection testing sample of smart mobile phone and the spectral wavelength of standard sample, light intensity, so, utilize smart mobile phone also may be used
To help experimenter to analyze the result of photometric detection experiment, and generate corresponding examining report, offer convenience to experimenter.
Wherein, this label is preferably two-dimension code label, naturally it is also possible to be the shape of other bar coded stickers and word tag
Formula.
Optionally, further comprise the steps of:
Form the photometric detection behaviour at least including detected sample information, standard sample information, sample type information, detection equipment
Make the cloud data base of method, sample detection methods, detectable information, operator's information；
And upload storage detect successfully record, failure logging and operational error record, detect time cumulation, detect sample, make
Used disposable device and operator, provide suggestion for operation accordingly.Use cloud data base that sample photometric detection is tested phase
Pass information carries out the renewal storage that prestores and follow up, and uses big data technique and cloud storage technology, analyzes and processes detection process
In details so that detection analyze convenient, be easier to successfully；And it is possible to test based on forefathers and previous photometric detection,
Follow-up luminosity test experience is instructed and advises, helps success and the progress of subsequent experimental.
Optionally, photometric detection operational approach at least includes two steps, uses smart mobile phone to use each step
Detection equipment and reagent carry out record；
Smart mobile phone, according to pre-setting, carries out image acquisition, and the figure that will collect to testing sample and standard sample automatically
The rgb value conversion of picture obtains spectral wavelength and light intensity.Detection equipment and reagent that different sample types use are not necessarily the same, right
It carries out record, subsequent experimental can be carried out directive function；Even and if same sample type, its detection equipment used and
Reagent, the most just the same, by relative analysis, the success more preferably faster of follow-up luminosity test experience can be helped and enter
Step；
It addition, have in the experiment of some photometric detection, the detection of the luminous intensity of testing result and absorbance etc. is had one
Excellent detection is interval, and autonomous setting gathers spectral wavelength, light intensity automatically, and experimenter can be helped the completeest
Become photometric detection experiment.
Optionally, if detected sample is pathogenic microorganism to be detected, standard sample is standard pathogenic microorganism；
Then according to corresponding photometric detection operational approach, pathogenic microorganism to be detected and standard pathogenic microorganism are carried out isothermal expansion
Increase detection；
To carrying out pathogenic microorganism to be detected and the standard pathogenic microorganism of isothermal duplication detection, carrying out image acquisition, it is right to obtain
The spectral wavelength answered and light intensity；
Obtain the luminosity information of pathogenic microorganism to be detected and standard pathogenic microorganism, and generate curve to be detected and the mark of correspondence
Directrix curve；
According to curve to be detected and the contrast of standard curve and analysis, obtain pathogenic microorganism to be detected and standard pathogenic microorganism
Photometric detection result；Certainly, this pathogenic microorganism to be detected can be one can also be multiple；
Concrete, the detection technique based on nucleic acid isothermal amplification technology has obtained swift and violent development in recent years.Nucleic acid isothermal
Amplification technique need not the time course of variations in temperature, has broken away from the dependence to superior instrument and equipment so that we are to pathogen
Checkout and diagnosis be able to quick and high-throughout realization.DNA is in dynamic balance state at about 65 DEG C, any one primer to
When the complementary portions of double-stranded DNA carries out base pairing extension, another chain will dissociate, become strand.Upstream internal primers F IP
F2 sequence be first combined with template F2c, under the effect of strand displacement type archaeal dna polymerase, with the 3' of FIP primers F 2 section end
End is starting point, matches with template DNA complementary series, starts strand displacement DNA synthesis.F3 primer and F2c front end F3c complementary,
With 3' end as starting point, by the effect of strand displacement type archaeal dna polymerase, replace the DNA of FIP primer synthesis ahead.One
Limit synthesizes self DNA, extends the most forward.The DNA that final F3 primer is synthesized into forms double-strand with template DNA.Drawn by FIP
The DNA that thing first synthesizes is carried out strand displacement by F3 primer and produces a strand, this strand 5' end exist complementary Flc and
, then there is oneself's base pairing, form circulus in Fl section.Meanwhile, BIP primer combines with the hybridization of this strand, with BIP
The 3' end of primer is starting point, synthesizes complementary strand, and circulus is opened in the process.Then, be similar to F3, B3 primer from
Insert outside BIP primer, carry out base pairing, with 3' end as starting point, under the effect of polymerase, synthesize new complementary strand.Logical
Cross above-mentioned two processes, form double-stranded DNA.And replaced single stranded DNA two ends exist complementary series, naturally-occurring oneself's base is joined
Right, form circulus, then whole piece chain presents dumbbell structure.This structure is the initial knot of LAMP method gene amplification circulation
Structure.All processes hereto are provided to be formed a dot structure of LAMP method gene amplification circulation.
In dumbbell structure, with the Fl section of 3' end as starting point, from as template, to carry out DNA synthesis and to extend.With
This simultaneously, FIP primers F 2 and strand F2c hybridization on ring, start new round strand replacement reaction.Dissociate the double-strand synthesized by F1 section
Nucleic acid.Equally, the single-chain nucleic acid dissociateed also can form circulus.Circulus exists single stranded form B2c,
B2 on BIP primer is hybrid with it, and starts new round amplification.Through identical process, form again circulus.By this mistake
Journey, result complementary series on same chain goes round and begins again and forms structure not of uniform size.
The method, and can be at isothermal mainly by 6 specific regions of 4 kinds of different specific primer identification target genes
Condition carries out amplified reaction.The amplification of gene and the detection of product can a step complete, and amplification efficiency is high, can expand at 15～60 min
Increase 109～1010 times；Specificity is high, and the detection of all target-gene sequences can having, without differentiating by amplified production.Have,
It is to utilize the fluorescence intensity of quantitative real time PCR Instrument detection reaction or utilize the burnt phosphorus produced in amplification process without amplified reaction
Acid magnesium precipitate reaction, precipitates what turbidity judged with transmissometer detection.
(1) fluorescent quantitation detection: utilize SYBR Green I fluorescent dye only and double-stranded DNA minor groove binding, when it with
During DNA double chain combination, sending the principle of the fluorescence of the strongest 800～1000 times, in an individual system, its signal intensity represents
The quantity of double chain DNA molecule.When nucleic acid synthesizes in a large number, SYBR Green I can mix double-stranded DNA automatically, by detection
Produced fluorescence intensity, obtains Ct value, reference standards curve, it may be determined that the starting copy number of template DNA.
(2) Turbidity measurement of magnesium pyrophosphate by-product: when nucleic acid synthesizes in a large number, the pyrophosphate ion separated out from dNTP
Mg2+ in reaction solution is combined, and produces magnesium pyrophosphate byproduct precipitate.Reaction equation is as follows:
(DNA)n-1 +dNTP=(DNA)n+P2O7 4-；P2O7 4-+2Mg2+=Mg2P2O7(precipitation)
This reaction has high specificity, under 400nm light, by photographic head detection precipitation turbidity just can interpolate that amplification with
No.
Optionally, in isothermal duplication detection, smart mobile phone control micro-fluidic LAMP chip to pathogenic microorganism to be detected and
Standard pathogenic microorganism carries out detection operation.Use smart mobile phone photographic head as detector, can identify any type of many
Channel chip, it is achieved effective differentiation of multiple LAMP signal, portable, low in energy consumption, simple to operate, the beneficially popularization of LAMP method
And application；Detection processes full-automation with data, and simple and convenient, layman also can Successful Operation；Use smart mobile phone figure
As processing module is as LAMP chip detector, can feature pathogen in automatization's Quantitative detection sample, have sensitive
Degree is high, detection speed is fast, high specificity, simple to operate, result is stable, automatically provide measurement result, the advantage such as portable.
Optionally, micro-fluidic LAMP chip is arranged on the chip carrier controlled by smart mobile phone, chip carrier be provided with for
The laser assembly in experimental optical source, and replaceable LAMP chip interface are provided；
Amplification pond it is provided with in micro-fluidic LAMP chip；It is provided with for detecting pathogenic microorganism to be measured and standard micro-in amplification pond
Biological specific probe.The replaceable LAMP chip interface that chip carrier uses, and interface has extensibility, can be according to reality
Border demand changes the chip of varying number sense channel, it is achieved the most multiple LAMP effectively expands and multiple LAMP detects signal
Effective differentiation, provide a kind of effective means for clinical multiple pathogens quick diagnosis；Use smart mobile phone as photoelectric colorimetry
Detector, smart mobile phone does not contacts with chip under test, thus avoids sample and chemical reagent to smart mobile phone
Corrosion, tester, also without the electric unit of contact chip, improves safety.
Optionally, if testing sample is test serum sample, standard sample is standard serum samples；
Then according to corresponding photometric detection operational approach, blood serum sample to be measured and standard serum samples are carried out serum glycated white egg
White detection；
Test serum sample after carrying out serum glycated albumin detection and standard sample are carried out image acquisition, obtains correspondence
Spectral wavelength and light intensity；
Obtain test serum sample and standard sample lower absorbance variable quantity under the light source of 570-650nm wavelength irradiates, and raw
Become corresponding curve to be measured and standard curve；
According to curve to be measured and the contrast of standard curve and analysis, obtain the glycosylated albumin content of test serum sample.At this
In photometric detection test, the absorbance of sample is directly proportional to the glycosylated albumin content of sample, by contrasting serum sample to be detected
Product and the absorbance curve of standard sample, can calculate the glycosylated albumin content of detected sample.
Concrete, in the assay method of (referenced patent CN104990879A) this serum glycated albumin content, hydrophilic
Property nano pipe/polyhenylethylene colloidal agent in add serum sample, albumin in sample and glycosylated albumin and existed by indifference absorption
Hydrophilic nano polystyrene colloid particle surface；It is subsequently adding special mouse-anti people's glycosylated albumin monoclonal antibody, is formed
Nano-particle-glycosylated albumin-mouse-anti people's glycosylated albumin mouse monoclonal antibody complex；This complex is white in sheep anti mouse saccharifying
Aggregation, cohesion amount and the absorption sugar at hydrophilic nano polystyrene colloid particle surface is produced under the effect of protein antibodies
Change albuminous amount to be directly proportional；Variable quantity according to measuring absorbance can be with glycosylated albumin institute accounting in blood serum sample to be detected
Rate.
Optionally, if testing sample is air sample to be measured, standard sample is normal air sample；
Then according to corresponding photometric detection operational approach, air sample to be measured and normal air sample are carried out PM2.5 particulate matter
Concentration Testing；
Air sample to be measured after carrying out the Concentration Testing of PM2.5 particulate matter and normal air sample are carried out image acquisition,
To corresponding spectral wavelength and light intensity；
Obtain air sample to be measured and the absorbance of normal air sample and generate curve to be measured and the standard curve of correspondence；
According to curve to be measured and the contrast of standard curve and analysis, obtain the concentration of the PM2.5 particulate matter of air sample to be measured.Sample
The concentration of the PM2.5 in product, the absorbance carrying out the product after photometric detection experiment to sample is relevant；So, in photometric detection
After experiment, by gathering air sample to be detected and the absorbance of normal air sample by mobile phone, and analyzer correspondence generates
Curve to be measured and standard curve, the most computable PM2.5 particle concentration to air sample to be detected.
Concrete, (referenced patent CN104865175A) first by PM2.5 and bigger particulate separation, by collect
50m3Air be stored in a big container, after tagging preserve.During detection, first scan air to be detected with smart mobile phone
Sample and the label of normal air sample, according to the prompting on mobile phone, scan incision device label, water scrubber label, reagent label
(buffer and developer), then according to the prompting on mobile phone, opens air pump, and air flows through sickle with certain flow velocity,
Bigger granule is retained by the parts being coated with oil because inertia is big, and the fine grained that inertia is less is most along with air stream
Pass through；Enter in water scrubber, then the fine particle of residual is scattered in the pure 30m through filtering3Distilled water in, add
PBS, developer uses citric acid (also can use ethylenediaminetetraacetic acid, ProcLin-300, or sodium thiosulfate) to enter with it
Row reaction, injects in color comparison tube.Scan each color comparison tube, starting hand-set photographic head and light source, detected by the system of the present invention
Go out the absorbance of standard solution and testing sample, and automatically testing sample is entered with the normal air sample concentration on working curve
Row compares, and obtains the concentration of PM2.5 particulate matter in air sample to be detected.
Optionally, if testing sample is chromium ion solution to be measured, standard sample is standard chlorine solion；
Then according to corresponding photometric detection operational approach, solution to be measured and standard solution are carried out content of chromium ion detection；
The solution to be measured and standard solution carrying out content of chromium ion detection is carried out image acquisition, obtain correspondence spectral wavelength and
Light intensity；
Obtain solution to be measured and the standard solution absorbance under the light source of 540nm wavelength irradiates and generate the curve to be measured of correspondence
And standard curve；
According to curve to be measured and the contrast of standard curve and analysis, obtain the content of chromium ion in solution to be detected.(with reference to specially
Profit CN104949932A) carry out photometric detection according to the detection operation method presentation on smart mobile phone, after standing the scheduled time, intelligence
Can automatically turn on by mobile phone camera, the absorbance at system display 540nm, generate corresponding curve to be measured and standard curve, i.e.
Can automatically calculate the content of chromium ion in sample.
Optionally, if detected sample is oils and fats to be detected, standard sample is standard oils and fats；
Then according to corresponding photometric detection operational approach, oils and fats to be detected and standard oils and fats are carried out Oxidation of Fat and Oils degree and examines
Survey；
Oils and fats to be detected and standard oils and fats are carried out image acquisition, obtains colourity and the intensity level of correspondence；
According to the oils and fats to be detected collected and the colourity of standard oils and fats and intensity level, display testing result also generates detection report
Accuse.Here obtaining of colourity and intensity level is relevant to spectral wavelength and light intensity.Scanning standard solution and solution to be detected
Label, by operation indicating, scans each reagent label, and adds reagent, and operates by given step；After colour developing, smart mobile phone is clapped
Take the photograph each test tube at the middle and upper levels or lower floor's color, according to the colourity identified and intensity level, automatically according to sample label and detection
Result generates curve to be measured and standard curve, and draws the concentration value of malonaldehyde in solution to be detected, thus judges this sample
The degree of oxidation of middle oils and fats.
Concrete, (referenced patent CN105004720A) is tested based on Kreis, i.e. Oxidation of Fat and Oils twice decomposition product and
The chromogenic reaction of benzenetriol, the present invention, by adjusting acid solution and the additional proportion of phloroglucinol solution, is greatly improved this anti-
The speed answered and color stability, overcome the defect that original experiment is unstable；It is greatly improved layered velocity, quickly obtains clarification thoroughly
Bright ratio chromatograph, reduces the interference of oil sample contrast color；Additionally can be substantially reduced the concentration of acid solution used, operate safer,
And reduce the garbage pollution to environment.
Generating carbonyl oxide during Oxidation of Fat and Oils, peroxide can do degree of oxidation and judge, but peroxide is in water
Deposit and the most easily decompose, therefore Oxidation of Fat and Oils is to after a certain degree, and peroxide value can reduce on the contrary.Therefore, peroxide
Should be the difference of existing amount of peroxides and decomposition amount, therefore other oxidimetry methods need to be coordinated, be beneficial to correctly sentencing of quality
Disconnected.
Kreis test is for aldehydes and the color reaction of ketone compounds, as the qualitative reaction of oils and fats whether oxidation deterioration,
Convenient and simple, use cost is low, and result is quick, is the most once becoming business, health supervision department is used primarily for evaluating
A kind of important method of inspection of fat oxidation.But the disadvantage of Kreis test is cannot be quantitative, causes the one of this reason
Individual key factor is, hardly results in consistent assay, different human users, or same personnel two between each laboratory
Secondary operation color there may be difference, so be only used as so far qualitative method use.The method of this invention can save base
Complex instrument during Kreis testing inspection, can quickly, the degree of oxidation of detection by quantitative oils and fats.
Scanned sample label, points out preparing standard solution according to mobile phone, and sets up label, and scanning standard solution label is pressed
Operation indicating, scans each reagent label, and adds reagent, and operates by given step.After colour developing, shoot each test tube with mobile phone
At the middle and upper levels or lower floor's color, according to the colourity identified and intensity level, generate automatically according to sample label and testing result and treat
Survey curve and standard curve, and draw the concentration value of malonaldehyde in solution to be detected, thus judge the oxygen of oils and fats in this sample
Change degree.
Above content is to combine concrete preferred implementation further description made for the present invention, it is impossible to assert
Being embodied as of the present invention is confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of present inventive concept, it is also possible to make some simple deduction or replace, all should be considered as belonging to the present invention's
Protection domain.
Claims (10)
1. a photometric detection method, it is characterised in that include step:
Smart mobile phone scanning testing sample and the label of standard sample；
Smart mobile phone is according to scanning the testing sample and the kind of standard sample obtained, the photometric detection operation side that display is corresponding
Method；
Smart mobile phone carries out the collection of image to the testing sample after processing according to photometric detection operational approach and standard sample；
It is scaled spectral wavelength and light intensity according to the image collected and is analyzed, calculates and processes, showing testing result
And generate examining report.
2. detection method as claimed in claim 1, it is characterised in that also include:
Form the photometric detection behaviour at least including detected sample information, standard sample information, sample type information, detection equipment
Make the cloud data base of method, sample detection methods, detectable information, operator's information；
And upload storage detect successfully record, failure logging and operational error record, detect time cumulation, detect sample, make
Used disposable device and operator, provide suggestion for operation.
3. detection method as claimed in claim 1, it is characterised in that described photometric detection operational approach at least includes two
Step, the detection equipment and the reagent that use smart mobile phone to use each step carry out record；
Smart mobile phone, according to pre-setting, carries out image acquisition, and the figure that will collect to testing sample and standard sample automatically
The rgb value conversion of picture obtains spectral wavelength and light intensity.
4. detection method as claimed in claim 1, it is characterised in that if detected sample is pathogenic microorganism to be detected, mark
Quasi-sample is standard pathogenic microorganism；
Then according to corresponding photometric detection operational approach, pathogenic microorganism to be detected and standard pathogenic microorganism are carried out isothermal expansion
Increase detection；
To carrying out pathogenic microorganism to be detected and the standard pathogenic microorganism of isothermal duplication detection, carry out spectral wavelength, light intensity
Gather；
Obtain the luminosity information of pathogenic microorganism to be detected and standard pathogenic microorganism, and generate curve to be detected and the mark of correspondence
Directrix curve；
According to curve to be detected and the contrast of standard curve and analysis, obtain pathogenic microorganism to be detected and standard pathogenic microorganism
Photometric detection result.
5. detection method as claimed in claim 4, it is characterised in that in described isothermal duplication detection, smart mobile phone controls
Micro-fluidic LAMP chip carries out detection operation to pathogenic microorganism to be detected and standard pathogenic microorganism.
6. detection method as claimed in claim 5, it is characterised in that described micro-fluidic LAMP chip is arranged on by described intelligence
On the chip carrier that mobile phone controls, described chip carrier is provided with the laser assembly for providing experimental optical source, and replaceable
LAMP chip interface；
Amplification pond it is provided with in described micro-fluidic LAMP chip；Amplification is provided with for detecting pathogenic microorganism to be measured and mark in pond
The specific probe of quasi-microorganism.
7. detection method as claimed in claim 1, it is characterised in that if testing sample is test serum sample, standard sample
For standard serum samples；
Then according to corresponding photometric detection operational approach, blood serum sample to be measured and standard serum samples are carried out serum glycated white egg
White detection；
Test serum sample after carrying out serum glycated albumin detection and standard sample are carried out image acquisition, obtains correspondence
Spectral wavelength and light intensity；
Obtain test serum sample and standard sample absorbance variable quantity under the light source of 570-650nm wavelength irradiates, and generate
Corresponding curve to be measured and standard curve；
According to curve to be measured and the contrast of standard curve and analysis, obtain the glycosylated albumin content of test serum sample.
8. detection method as claimed in claim 1, it is characterised in that if testing sample is air sample to be measured, standard sample
For normal air sample；
Then according to corresponding photometric detection operational approach, air sample to be measured and normal air sample are carried out PM2.5 particulate matter
Concentration Testing；
Air sample to be measured after carrying out the Concentration Testing of PM2.5 particulate matter and normal air sample are carried out image acquisition,
To corresponding spectral wavelength and light intensity；
Obtain air sample to be measured and the absorbance of normal air sample and generate curve to be measured and the standard curve of correspondence；
According to curve to be measured and the contrast of standard curve and analysis, obtain the concentration of the PM2.5 particulate matter of air sample to be measured.
9. detection method as claimed in claim 1, it is characterised in that if testing sample is chromium ion solution to be measured, standard sample
Product are standard chlorine solion；
Then according to corresponding photometric detection operational approach, solution to be measured and standard solution are carried out content of chromium ion detection；
The solution to be measured and standard solution carrying out content of chromium ion detection is carried out image acquisition, obtains the spectrum ripple of correspondence
Length and light intensity；
Obtain solution to be measured and the standard solution absorbance under the light source of 540nm wavelength irradiates and generate the curve to be measured of correspondence
And standard curve；
According to curve to be measured and the contrast of standard curve and analysis, obtain the content of chromium ion in solution to be detected.
10. detection method as claimed in claim 1, it is characterised in that if detected sample is oils and fats to be detected, standard sample
For standard oils and fats；
Then according to corresponding photometric detection operational approach, oils and fats to be detected and standard oils and fats are carried out Oxidation of Fat and Oils degree and examines
Survey；
Oils and fats to be detected and standard oils and fats are carried out image acquisition, obtains colourity and the intensity level of correspondence；
According to the oils and fats to be detected collected and the colourity of standard oils and fats and intensity level, display testing result also generates detection report
Accuse.
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|---|---|---|---|
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