CN106591443A - Method for analytical processing of crop fluorescence capillary electrophoretogram - Google Patents
Method for analytical processing of crop fluorescence capillary electrophoretogram Download PDFInfo
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Abstract
The invention relates to a method and device for analytical processing of a crop fluorescence capillary electrophoretogram, wherein the method includes 3 major processing steps of curve processing, internal standard calibration, and fragment analysis and genotyping, and is characterized in that curve processing includes pull-up peak processing, the pull-up peak processing includes the steps of overlapped peak positioning, overlapped peak grouping, overlapped peak processing, quasi pull-up peak grouping, quasi pull-up peak re-grouping, pull-up peak III judging and pull-up peak III processing. The device includes three modules of curve processing, internal standard calibration, and fragment analysis and genotyping and is characterized in that the curve processing module includes a pull-up peak processing sub module. The problem of internal standard deviation caused because a peak at an internal standard peak position is deleted can be effectively avoided, the sequencing accuracy rate of the crop fluorescence capillary electrophoretogram is improved significantly, and multiple laboratories are facilitated to work together to construct a crop genetic information fingerprint database.
Description
Technical field
This invention relates generally to biogenetics and genomics field, and more specifically it relates at a kind of analysis
The method and its device of reason crops fluorescent capillary electrophoresis tube collection of illustrative plates.
Background technology
Fluorescent capillary electrophoresis tube is the conventional method of Treatment Analysis bio-genetic material.The genetic fragments of different length can be with
It is separated by electrophoresis, and the genetic fragments of different but identical length of originating then can be made a distinction by different fluorescence colors.
When fluorescent dye primer is used, the presence or absence of pcr amplification product or how much by fluorescence intensity judging, but it is different
There is overlap between the fluorescing frequencies of color.If the overflow of a certain color, has exceeded the process range of program, or
Also there is the small peak of another kind of color in power of test of the person beyond instrument, the position that superelevation peak will occur in a kind of color,
This small peak is and pulls up (pull up) peak.It is this as color is diffused into " pulling up existing for another from spectrum channel
As " it is widely present in fluorescence electrophoresis experimental situation.
Relative to the research of mankind's inheritance resources, the research of crops genetic resourcess falls behind relatively.Currently used for crops
The design of primers of genetic marker is relatively deficient, the test kit for also not having maturation, and this causes Jing PCR amplifications and fluorescent capillary electrophoresis tube
There is the more peak that pulls up in the crops gene test collection of illustrative plates of generation, and these pull up peak peak height it is higher, it is not easy to other
Distinguish at normal peak.Also, due to reference material produce internal standard colour band peak height it is relatively low, if now sample produce common color
Band has the peak for overlapping with internal standard colour band, and normal internal standard base peak will likely be taken as pulling up peak and being deleted, so as to meeting
Cause internal standard calibration poor or failure.Such situation is pulled up in crops can be tighter under the higher environment of the universal peak height in peak
Weight.In addition, the gene test collection of illustrative plates produced after crops electrophoresis easily produces continuous multimodal, i.e., a series of equidistant, spacing are not
More than 2.5bp and peak number more than 2 peak, if pulling up peak occurs in continuous multimodal region, it is likely that continuous multimodal can be taken as
By and be ignored, cause more to read peaks or misread the situation at peak.
It is existing process fluorescent capillary electrophoresis tube collection of illustrative plates in pull up peak method be generally only by adjacent bands collection of illustrative plates contrast with
Examination goes out to pull up peak, and the peak that pulls up that examination is gone out simply is deleted.But this method cannot effective process farming analyte detection collection of illustrative plates
In have with internal standard colour band and overlap or pull up peak in continuous many crest segments, often so that internal standard calibration is poor in addition failure or
Cause many reading peaks or misread the situation at peak, failure is even detected so as to cause testing result relatively large deviation occur.Therefore, how
Peak can will be pulled up in the case where internal standard calibration and loss peak is not affected and do effective process, so as to improve crops fluorescent capillary pipe
Electrophoresis pattern gene test accuracy is the key technical problem that those skilled in the art need to solve.
The content of the invention
Asking for peak cannot be pulled up in effective process crops fluorescent labeling detection collection of illustrative plates to solve to exist in prior art
Topic, it is it is an object of the invention to provide a kind of method that analyzing and processing crops fluorescent labeling detects collection of illustrative plates including curve processing, interior
Calibration standard and 3 main processing steps of fragment analysis and gene type, it is characterised in that also include pulling up peak in curve processing
Process, it is described pull up peak process comprise the following steps：
1) coincidence peak positioning：A colour bands are contrasted with B colour bands, in finding B colour bands, position is overlapped with the position at A colour bands peak
Peak, these peaks are set to into coincidence peak, wherein A colors have Chong Die and A colour bands that B colour bands can be caused to pull up with B color fluorescence spectruies；
2) coincidence peak packet：
When step 1) position at coincidence peak that positions when being overlapped with the position of internal standard base peak, these coincidence peaks is set to and pulled out
Peak I is played,
When coincidence peak is located at continuous many crest segments, these coincidence peaks for being located at continuous many crest segments is set to into standard and pulls up peak；
When the position at coincidence peak is not neither overlapped with internal standard base peak in continuous many crest segments again, these coincidence peaks are set to
Peak II is pulled up,
3) coincidence peak is processed：
Record each position for pulling up peak I peak I is pulled up so that follow-up internal standard calibration steps is referred to and eliminated,
Elimination pulls up peak II,
Record standard pulls up the position X that total number N and each standard at peak pulls up peaki B, wherein, i is no less than 0 and is not more than
The integer of N,
When N is 0, end pulls up peak process step；
4) standard pulls up peak packet：It is X by positioni BPeak in all B colour bands of ± 32frame is divided into i-th group；
5) standard pulls up peak and is grouped again：N number of group in step 4 is grouped again, that is, is found per the summit in group, will most
Peak and its two peaks of left and right are divided into subgroup 1i of this group, and other peaks in group are divided into another subgroup 2i；
6) peak III judgements are pulled up：In each ai subgroup that step 5 determines,
A) peak height at A colour bands peak in the range of subgroup position is sorted and is set to the position of the summit in the range of this
Xai A, the position of summit in subgroup is set to into X in additionai B,
B) by Xai BWith Xai BContrast, if Xai B≠Xai B, all peaks in subgroup ai are all judged as not being to pull up peak III,
Otherwise next step judgement is then carried out,
C) when position is Xai BPeak peak height Hai BIt is X with positionai APeak peak height Hai ABetween ratio be more than c when,
Position is Xai BPeak be judged to be defined and pull up peak III, the wherein value of c is adjusted according to tested species,
D) sort from high to low according to peak height, judge that other peaks are in subgroup ai according to the height ratio of the position and peak at peak
No being defined pulls up peak III,
E) when the ratio of the number at all peaks in the number and subgroup ai that standard in subgroup ai pulls up peak III is less than 0.75
When, all peaks in subgroup ai are all judged as not being to pull up peak III, otherwise are all then to pull up peak III,
Wherein a is subgroup number, and a values are 1 or 2；
7) pull up peak III process：Peak III eliminations will be pulled up, and terminates behind to pull up peak process step.
Further, step 7 pull up peak III process described in elimination be：The adjacent situations of peak III are pulled up when existing
When, adjacent two pull up peak III and adopt curve difference approximating method to be combined process and pull up peak III to eliminate, otherwise then
Carry out unimodal process peak III is pulled up to eliminate.Can be made Jing pulling up after peak III Processing for removing using curve difference approximating method
Picture is more smoothed, it is to avoid the figures such as spine is occurred and is produced impact to follow-up atlas analysis.
Further, the curve difference approximating method is linear interpolation or cubic spline interpolation.Imitate in same process
Under fruit, linear interpolation and cubic spline interpolation graphics calculations amount are less, and graphics process speed is faster.
Further, the curve processing also include nail peak process step, nail peak process step be by due to
During electrophoresis, electric current is unstable and caused nail-like spike is removed.
Further, the curve processing also include saturation peak process step, saturation peak process step be by due to
Peak intensity has exceeded the threshold value of sequenator and caused tack or concave head type saturation peak and the pulling out of occurring of correspondence under saturation peak
Play peak superimposed so as to form a complete peak.
It is another object of the present invention to be to provide the device that a kind of analyzing and processing crops fluorescent labeling detects collection of illustrative plates, institute
Stating device includes curve processing, internal standard calibration and 3 modules of fragment analysis and gene type, it is characterised in that curve processing
Also include in module that pulling up peak processes submodule, the peak processing module that pulls up is included with lower unit：
Coincidence peak positioning unit, for A colour bands are contrasted with B colour bands, finds the position at position and A colour bands peak in B colour bands
The peak of coincidence is put, these peaks are set to into coincidence peak, wherein A colors have Chong Die with B color fluorescence spectruies and A colour bands can cause B colour bands
Pull up；
Coincidence peak grouped element, for being grouped to coincidence peak, specially：
When the position for being set to the coincidence peak positioned by unit is overlapped with internal standard standard peak position, these are overlapped by this unit
Peak is set to and pulls up peak I,
When the position at coincidence peak is not neither overlapped with internal standard base peak in continuous many crest segments again, these are overlapped by this unit
Peak is set to and pulls up peak II,
When coincidence peak is located at continuous many crest segments, these coincidence peaks for being located at continuous many crest segments are set to standard and are pulled up by this unit
Peak；
Coincidence peak processing unit, for processing the Chong Hefeng after packet respectively, specially：
Record each position for pulling up peak I peak I is pulled up so that follow-up internal standard calibration module is referred to and eliminated,
Elimination pulls up peak II,
Record standard pulls up the position X that total number N and each standard at peak pulls up peaki B, wherein, i is no less than 0 and is not more than
The integer of N,
When N is 0, end pulls up peak process；
Standard pulls up peak grouped element, pulls up peak for alignment and is grouped, specially：It is X by positioni B± 32frame's
Peak in all B colour bands is divided into i-th group；
Standard pulls up peak grouped element again, pulls up peak for alignment and is further grouped, specially：It is single that standard pulls up peak packet
N number of group after unit is processed is grouped again, that is, find per the summit in group, and summit and its two peaks of left and right are divided into this group
Subgroup 1i, other peaks in group are divided into another subgroup 2i；
Peak III judging units are pulled up, and the judgement at peak are pulled up for alignment, specially：Standard pulls up peak, and grouped element determines again
Each ai subgroup in,
A) peak height at the peak of A colour bands in the range of subgroup position is sorted and is set to the position of the summit in the range of this
Xai A, the position of summit in subgroup is set to into X in additionai B,
B) by Xai BWith Xai BContrast, if Xai B≠Xai B, all peaks in subgroup ai are all judged as not being to pull up peak III,
Otherwise next step judgement is then carried out,
C) when position is Xai BPeak peak height Hai BIt is X with positionai APeak peak height Hai ABetween ratio be more than c when,
Position is Xai BPeak be judged as pulling up peak III, the wherein value of c is adjusted according to tested species,
D) judge whether other peaks are to pull up peak III in subgroup ai according to the height of the position and peak at peak,
E) when the ratio of the number at all peaks in number and subgroup ai that peak III is pulled up in subgroup ai is less than 0.75,
All peaks in subgroup ai are all judged as not being to pull up peak III, otherwise are all then to pull up peak III,
Wherein a is subgroup number, and a values are 1 or 2；
Peak III processing units are pulled up, for peak III eliminations will be pulled up, and terminates behind to pull up peak process.
Further, the curve processing module also includes that nail peak processes submodule, and the nail peak processes submodule
For will due to during electrophoresis electric current it is unstable and caused nail-like spike is removed.
Further, the curve processing module also includes that saturation peak processes submodule, and the saturation peak processes submodule
For will exceed the threshold value of sequenator due to peak intensity and caused tack or concave head type saturation peak and under saturation peak it is right
Should occur to pull up peak superimposed so as to form a complete peak.
It is a further object of the present invention to provide a kind of electric terminal, it is characterised in that the electric terminal includes above-mentioned
Any one analyzing and processing fluorescent labeling detects the device of collection of illustrative plates.
Further, the electric terminal also includes：
Communication module, for by way of wiredly and/or wirelessly by the analyzing and processing crops SSR marker collection of illustrative plates
Collection of illustrative plates or the corresponding data transfer of collection of illustrative plates after device process is to outside other equipment.
Curve processing of the present invention may also include the means such as baseline calibration, curve smoothing process.
Due to the interference that there is fluorescent material in electrophoresis system, the baseline of each colour band in collection of illustrative plates there may be deviation.
Baseline calibration of the present invention is that the minimum peak of each colour band is set to 0, makes each colour band on same datum line.
Smoothing processing is to be smoothed zigzag fashion present in collection of illustrative plates, becomes apparent from original peaks readable.
Due to the interference that there is operating error and other materials in PCR amplifications and electrophoretic process, SSR collection of illustrative plates there may be
The situation of horizontal displacement.Internal standard calibration steps of the present invention is will to detect internal standard substance in obtained collection of illustrative plates (also known as reference material)
The position at corresponding peak is compared with the position at each peak in internal standard substance standard diagram, to calibrate the abscissa of collection of illustrative plates, that is, is migrated
Distance (frame).
Fragment analysis (GeneScan) of the present invention are that the internal standard substance of known length is done molecular weight standard first to obtain
Standard curve of the mrna length (bp) to migration distance (frame), the peak of the unknown sample of detection and standard curve are compared,
Calculate the corresponding actual gene fragment length (bp) in each peak.
Gene type (Genotyping) of the present invention is by the peak and allelic ladder of unknown sample
(allelic ladder) compares, to confirm the corresponding allele in peak.
The peak number mentioned in the present invention refers to the number at peak.
Method compared with prior art excellent of analyzing and processing crops fluorescent capillary electrophoresis tube collection of illustrative plates of the present invention
Put and be：As the peak that pulls up to diverse location is made and being handled differently, peak can be pulled up in effectively elimination and spectrum recognition is caused
Impact while can also be prevented effectively from continuous multimodal be directly deleted so as to cause hereditary information omit problem, can also have
Effect avoids the internal standard calibration offset issue for being deleted and being caused due to the peak of internal standard standard peak position, makes crops fluorescent capillary pipe
The sequencing accuracy rate of electrophoresis pattern is obviously improved, and is conducive to many laboratorys to cooperate jointly and is built crops hereditary information fingerprint
Storehouse.
Description of the drawings
Fig. 1 is the part detection collection of illustrative plates contrast of an embodiment of the method through the present invention and a method comparative example process
Figure, wherein upper half figure are the part detection collection of illustrative plates processed through the comparative example, and lower half figure is the same part that the embodiment is processed
Detection collection of illustrative plates；
Fig. 2 is the functional block diagram of a device embodiment of the present invention；
Fig. 3 is the functional block diagram of another device embodiment of the present invention.
Specific embodiment
Embodiment of the method 1
Be sequenced using the SSR marker of P40 primer pair Semen Maydiss, and corresponding detection is formed using PCR amplifications and fluorescent labeling
Collection of illustrative plates.
Process is analyzed to the detection collection of illustrative plates, the analyzing and processing includes curve processing, internal standard calibration and fragment point
Analysis and 3 main processing steps of gene type, it is characterised in that also include pulling up peak process in curve processing, it is described to pull up peak
Process is comprised the following steps：
1) coincidence peak positioning：A colour bands are contrasted with B colour bands, in finding B colour bands, position is overlapped with the position at A colour bands peak
Peak, these peaks are set to into coincidence peak, wherein A colors have Chong Die and A colour bands that B colour bands can be caused to pull up with B color fluorescence spectruies；
2) coincidence peak packet：
When step 1) position at coincidence peak that positions when being overlapped with the position of internal standard base peak, these coincidence peaks is set to and pulled out
Peak I is played,
When coincidence peak is located at continuous many crest segments, these coincidence peaks for being located at continuous many crest segments is set to into standard and pulls up peak；
When the position at coincidence peak is not neither overlapped with internal standard base peak in continuous many crest segments again, these coincidence peaks are set to
Peak II is pulled up,
3) coincidence peak is processed：
Record each position for pulling up peak I peak I is pulled up so that follow-up internal standard calibration steps is referred to and eliminated,
Elimination pulls up peak II,
Record standard pulls up the position X that total number N and each standard at peak pulls up peaki B, wherein, i is no less than 0 and is not more than
The integer of N,
When N is 0, end pulls up peak process step；
4) standard pulls up peak packet：It is X by positioni BPeak in all B colour bands of ± 32frame is divided into i-th group；
5) standard pulls up peak and is grouped again：N number of group in step 4 is grouped again, that is, is found per the summit in group, will most
Peak and its two peaks of left and right are divided into subgroup 1i of this group, and other peaks in group are divided into another subgroup 2i；
6) peak III judgements are pulled up：In each ai subgroup that step 5 determines,
A) peak height at A colour bands peak in the range of subgroup position is sorted and is set to the position of the summit in the range of this
Xai A, the position of summit in subgroup is set to into X in additionai B,
B) by Xai BWith Xai BContrast, if Xai B≠Xai B, all peaks in subgroup ai are all judged as not being to pull up peak III,
Otherwise next step judgement is then carried out,
C) when position is Xai BPeak peak height Hai BIt is X with positionai APeak peak height Hai ABetween ratio be more than c when,
Position is Xai BPeak be judged to be defined and pull up peak III, the wherein value of c is adjusted according to tested species,
D) sort from high to low according to peak height, judge that other peaks are in subgroup ai according to the height ratio of the position and peak at peak
No being defined pulls up peak III,
E) when the ratio of the number at all peaks in the number and subgroup ai that standard in subgroup ai pulls up peak III is less than 0.75
When, all peaks in subgroup ai are all judged as not being to pull up peak III, otherwise are all then to pull up peak III,
Wherein a is subgroup number, and a values are 1 or 2；
7) pull up peak III process：Peak III eliminations will be pulled up, and terminates behind to pull up peak process step.
Detection collection of illustrative plates after process see accompanying drawing 1 (under).
Method comparative example 1
Be sequenced using the SSR marker of P40 primer pair Semen Maydiss, and corresponding detection is formed using PCR amplifications and fluorescent labeling
Collection of illustrative plates.
Be analyzed process to the detection collection of illustrative plates, the analyzing and processing in addition to not pulling up peak and processing, remaining and enforcement
Treatment Analysis described in example 1 are identical.
Detection collection of illustrative plates after process see accompanying drawing 1 (on).
Can be seen by accompanying drawing 1, carrying out pulling up after peak processes, B colour bands (lower section colour band) 298frame nearby with
310frame is nearby eliminated by the peak that pulls up that the continuous multimodal of A colour bands takes up, in addition, the peak of the corresponding continuous multimodal of A colour bands
Height is corrected, and is conducive to follow-up other image procossings.
Experimental example 1
Sequencing 3 times are repeated with sample sequencing point to 200 known arrays of corn and soybean, Semen Tritici aestivi, it is same to every kind of crops
The sequencing procedure of one known array is identical.Sequencing uses SSR marker, PCR amplifications and fluorescent labeling to form detection collection of illustrative plates, Mei Zhongnong
Crop respectively obtains 300 detection collection of illustrative plates.By these collection of illustrative plates respectively through process (i.e. similar to Example 1) and discontinuous multimodal
Processing method (i.e. similar to comparative example 1) is analyzed process, and the result after analyzing and processing and known array are contrasted, and obtains
To the average detected error rate of each method, concrete outcome see the table below.
After the process of continuous multimodal, detect that the average detected error rate of collection of illustrative plates is substantially reduced, substantially improve farming
The sequencing accuracy of thing fluorescent capillary electrophoresis tube collection of illustrative plates, is conducive to many laboratorys to cooperate jointly and builds crops hereditary information fingerprint
Storehouse.
Fig. 2 is the functional block diagram of a device embodiment of the present invention.As shown in Fig. 2 the device 100 includes：
Including curve processing module 110, internal standard calibration module 120, fragment analysis and gene type module 130, at curve
Also include in reason that pulling up peak processes submodule 111, the peak process submodule that pulls up is included with lower unit：
Coincidence peak positioning unit 1111, for A colour bands are contrasted with B colour bands, finds position and A colour bands peak in B colour bands
Position overlap peak；
Coincidence peak grouped element 1112, for being grouped to coincidence peak；
Coincidence peak processing unit 1113, for processing the Chong Hefeng after packet respectively；
Standard pulls up peak grouped element 1114, pulls up peak for alignment and is grouped；
Standard pulls up peak grouped element 1115 again, pulls up peak for alignment and is further grouped；
Peak III judging unit 1116 is pulled up, and the judgement at peak is pulled up for alignment；
Peak III processing unit 1117 is pulled up, for peak III eliminations will be pulled up.
Fig. 3 is the functional block diagram of a device embodiment of the present invention.As shown in figure 3, the curve processing in the device 100
Module 110 can also include that nail peak processes submodule 112, for will as during electrophoresis, electric current is unstable caused nail-like point
Peak is removed；And saturation peak processes submodule 113, for the threshold value of sequenator will have been exceeded due to peak intensity and caused tack
Or concave head type saturation peak and under saturation peak correspondence occur to pull up peak superimposed so as to form a complete peak.
The method of work of the device of the embodiment of the present invention please refer to above method embodiment, therefore will not be described in detail herein.
The embodiment of the present invention also provides a kind of electric terminal, and the electric terminal includes that the analyzing and processing crops of the above are glimmering
The device of light capillary electrophoresis collection of illustrative plates.Further, the electric terminal can also include：Communication unit, for by wired
And/or the collection of illustrative plates or collection of illustrative plates after wirelessly processed the device of the analyzing and processing crops fluorescent capillary electrophoresis tube collection of illustrative plates
Corresponding data transfer is to outside other equipment.
Through the above description of the embodiments, those skilled in the art can be understood that the present invention can be by
Software adds the mode of required general hardware platform to realize, naturally it is also possible to by hardware, or the combination of the two is implementing.
Based on such understanding, the part that technical scheme is substantially contributed to prior art in other words can be with software
The form of product is embodied, and the software module or computer software product can be stored in a storage medium, if including
Dry instruction is used so that a computer equipment (can be personal computer, server, or network equipment etc.) performs this
Method described in bright each embodiment.Storage medium can be random access memory (RAM), internal memory, read only memory (ROM), electricity
Well known in programming ROM, electrically erasable ROM, depositor, hard disk, moveable magnetic disc, CD-ROM or technical field
Any other form of storage medium.
Although the principle of the present invention has had specific descriptions herein, it will be appreciated by those of skill in the art that this
Individual description only by way of example carrying out, and not as the restriction with regard to the scope of the present invention.Except shown herein
Outside the exemplary embodiment of description, within the scope of the invention by the modification and replacement of one of ordinary skill in the art
Other embodiment, be regarded as within the scope of the invention.
Claims (10)
1. a kind of method that analyzing and processing crops fluorescent labeling detects collection of illustrative plates, including the calibration of curve processing, internal standard and fragment
Analysis and 3 main processing steps of gene type, it is characterised in that also include pulling up peak process in curve processing, it is described to pull up
Peak process is comprised the following steps：
1) coincidence peak positioning：A colour bands are contrasted with B colour bands, the peak that position in B colour bands is overlapped with the position at A colour bands peak is found,
These peaks are set to into coincidence peak, wherein A colors there are Chong Die and A colour bands that B colour bands can be caused to pull up with B color fluorescence spectruies；
2) coincidence peak packet：
When step 1) position at coincidence peak that positions when being overlapped with the position of internal standard base peak, these coincidence peaks is set to and pulls up peak
I,
When coincidence peak is located at continuous many crest segments, these coincidence peaks for being located at continuous many crest segments is set to into standard and pulls up peak；
When the position at coincidence peak is not neither overlapped with internal standard base peak in continuous many crest segments again, these coincidence peaks are set to and are pulled up
Peak II,
3) coincidence peak is processed：
Record each position for pulling up peak I peak I is pulled up so that follow-up internal standard calibration steps is referred to and eliminated,
Elimination pulls up peak II,
Record standard pulls up the position X that total number N and each standard at peak pulls up peaki B, wherein, i is no less than 0 and no more than N
Integer,
When N is 0, end pulls up peak process step；
4) standard pulls up peak packet：It is X by positioni BPeak in all B colour bands of ± 32frame is divided into i-th group；
5) standard pulls up peak and is grouped again：N number of group in step 4 is grouped again, that is, is found per the summit in group, by summit
And its two peaks of left and right are divided into subgroup 1i of this group, other peaks in group are divided into another subgroup 2i；
6) peak III judgements are pulled up：In each ai subgroup that step 5 determines,
A) peak height at A colour bands peak in the range of subgroup position is sorted and the position of the summit in the range of this is set to into Xai A, separately
The outer position by summit in subgroup is set to Xai B,
B) by Xai BWith Xai BContrast, if Xai B≠Xai B, all peaks in subgroup ai are all judged as not being to pull up peak III, otherwise
Next step judgement is carried out then,
C) when position is Xai BPeak peak height Hai BIt is X with positionai APeak peak height Hai ABetween ratio be more than c when, position
For Xai BPeak be judged to be defined and pull up peak III, the wherein value of c is adjusted according to tested species,
D) sort from high to low according to peak height, judge in subgroup ai that whether other peaks are according to the height ratio of the position and peak at peak
Standard pulls up peak III,
E) when the ratio of the number at all peaks in number and subgroup ai that standard in subgroup ai pulls up peak III is less than 0.75, son
Group ai in all peaks be all judged as not being to pull up peak III, otherwise be all then to pull up peak III,
Wherein a is subgroup number, and a values are 1 or 2；
7) pull up peak III process：Peak III eliminations will be pulled up, and terminates behind to pull up peak process step.
2. method according to claim 1, it is characterised in that step 7 pull up peak III process described in elimination be：When
When presence pulls up peak III adjacent situations, adjacent two pull up peak III and are combined process using curve difference approximating method
Peak III is pulled up to eliminate, otherwise is then carried out unimodal process and peak III is pulled up to eliminate.
3. method according to claim 2, it is characterised in that：The curve difference approximating method is linear interpolation or three times
Spline interpolation.
4. the method according to any one of claim 1-3, it is characterised in that：The curve processing is also included at nail peak
Reason step, nail peak process step are by due to electric current shakiness during electrophoresis, caused nail-like spike is removed.
5. the method according to any one of claim 1-4, it is characterised in that：The curve processing is also included at saturation peak
Reason step, saturation peak process step are will to have exceeded the threshold value of sequenator due to peak intensity and caused tack or concave head type
Saturation peak and under saturation peak correspondence occur to pull up peak superimposed so as to form a complete peak.
6. a kind of analyzing and processing crops fluorescent labeling detects the device of collection of illustrative plates, and described device includes curve processing, internal standard calibration
And 3 modules of fragment analysis and gene type, it is characterised in that also include in curve processing module that pulling up peak processes submodule
Block, the peak processing module that pulls up are included with lower unit：
Coincidence peak positioning unit, for A colour bands are contrasted with B colour bands, in finding B colour bands, position is heavy with the position at A colour bands peak
These peaks are set to coincidence peak by the peak of conjunction, and wherein A colors have Chong Die and A colour bands that B colour bands can be caused to pull up with B color fluorescence spectruies；
Coincidence peak grouped element, for being grouped to coincidence peak, specially：
When the position for being set to the coincidence peak positioned by unit is overlapped with internal standard standard peak position, this unit is fixed by these coincidence peaks
To pull up peak I,
When the position at coincidence peak is not neither overlapped with internal standard base peak in continuous many crest segments again, this unit is fixed by these coincidence peaks
To pull up peak II,
When coincidence peak is located at continuous many crest segments, these coincidence peaks for being located at continuous many crest segments are set to standard and pull up peak by this unit；
Coincidence peak processing unit, for processing the Chong Hefeng after packet respectively, specially：
Record each position for pulling up peak I peak I is pulled up so that follow-up internal standard calibration module is referred to and eliminated,
Elimination pulls up peak II,
Record standard pulls up the position X that total number N and each standard at peak pulls up peaki B, wherein, i is no less than 0 and no more than N
Integer,
When N is 0, end pulls up peak process；
Standard pulls up peak grouped element, pulls up peak for alignment and is grouped, specially：It is X by positioni BAll B of ± 32frame
Peak in colour band is divided into i-th group；
Standard pulls up peak grouped element again, pulls up peak for alignment and is further grouped, specially：Standard is pulled up at peak grouped element
N number of group after reason is grouped again, that is, find per the summit in group, and summit and its two peaks of left and right are divided into the one of this group
Individual subgroup 1i, other peaks in group are divided into another subgroup 2i；
Peak III judging units are pulled up, and the judgement at peak are pulled up for alignment, specially：Standard pull up peak again grouped element determine it is every
In individual ai subgroups,
A) peak height at the peak of A colour bands in the range of subgroup position is sorted and the position of the summit in the range of this is set to into Xai A,
The position of summit in subgroup is set to into X in additionai B,
B) by Xai BWith Xai BContrast, if Xai B≠Xai B, all peaks in subgroup ai are all judged as not being to pull up peak III, otherwise
Next step judgement is carried out then,
C) when position is Xai BPeak peak height Hai BIt is X with positionai APeak peak height Hai ABetween ratio be more than c when, position
For Xai BPeak be judged as pulling up peak III, the wherein value of c is adjusted according to tested species,
D) judge whether other peaks are to pull up peak III in subgroup ai according to the height of the position and peak at peak,
E) when the ratio of the number at all peaks in number and subgroup ai that peak III is pulled up in subgroup ai is less than 0.75, subgroup
All peaks in ai are all judged as not being to pull up peak III, otherwise are all then to pull up peak III,
Wherein a is subgroup number, and a values are 1 or 2；
Peak III processing units are pulled up, for peak III eliminations will be pulled up, and terminates behind to pull up peak process.
7. device according to claim 6, it is characterised in that：The curve processing module also includes that nail peak processes submodule
Block, the nail peak process submodule is used for will the caused nail-like spike removal due to electric current shakiness during electrophoresis.
8. device according to claim 6, it is characterised in that：The curve processing module also includes that saturation peak processes submodule
Block, the saturation peak process submodule for having exceeded the threshold value of sequenator due to peak intensity and caused tack or concave head type
Saturation peak and under saturation peak correspondence occur to pull up peak superimposed so as to form a complete peak.
9. a kind of electric terminal, it is characterised in that the electric terminal is included at the analysis any one of claim 4-6
Reason crops fluorescent labeling detects the device of collection of illustrative plates.
10. electric terminal according to claim 7, it is characterised in that the electric terminal also includes：
Communication module, for the analyzing and processing crops fluorescent labeling is detected collection of illustrative plates by way of wiredly and/or wirelessly
Device process after collection of illustrative plates or the corresponding data transfer of collection of illustrative plates to outside other equipment.
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| CN102880812A (en) * | 2012-08-02 | 2013-01-16 | 北京华生恒业科技有限公司 | Method and device for processing solubility curve based on wavelet analysis method, and electronic terminal |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102880812A (en) * | 2012-08-02 | 2013-01-16 | 北京华生恒业科技有限公司 | Method and device for processing solubility curve based on wavelet analysis method, and electronic terminal |
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| MITCHELL M. HOLLAND 等: "《GeneMarker_ HID: A Reliable Software Tool for the Analysis of Forensic STR Data》", 《J FORENSIC SCI》 * |
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