CN114295836A - Computing equipment, storage medium and glioma IDH typing detection device and system - Google Patents
Computing equipment, storage medium and glioma IDH typing detection device and system Download PDFInfo
- Publication number
- CN114295836A CN114295836A CN202111586578.6A CN202111586578A CN114295836A CN 114295836 A CN114295836 A CN 114295836A CN 202111586578 A CN202111586578 A CN 202111586578A CN 114295836 A CN114295836 A CN 114295836A
- Authority
- CN
- China
- Prior art keywords
- peak intensity
- glioma
- acid peak
- idh
- glutamine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 208000032612 Glial tumor Diseases 0.000 title claims abstract description 97
- 206010018338 Glioma Diseases 0.000 title claims abstract description 97
- 238000001514 detection method Methods 0.000 title claims abstract description 37
- HWXBTNAVRSUOJR-UHFFFAOYSA-N alpha-hydroxyglutaric acid Natural products OC(=O)C(O)CCC(O)=O HWXBTNAVRSUOJR-UHFFFAOYSA-N 0.000 claims abstract description 126
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims abstract description 47
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims abstract description 45
- 235000013922 glutamic acid Nutrition 0.000 claims abstract description 39
- 239000004220 glutamic acid Substances 0.000 claims abstract description 39
- 238000001819 mass spectrum Methods 0.000 claims abstract description 23
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 50
- 238000005507 spraying Methods 0.000 claims description 25
- 239000011159 matrix material Substances 0.000 claims description 18
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 claims description 8
- 229930195712 glutamate Natural products 0.000 claims description 6
- 239000007921 spray Substances 0.000 claims description 6
- 108010075869 Isocitrate Dehydrogenase Proteins 0.000 description 59
- 102000012011 Isocitrate Dehydrogenase Human genes 0.000 description 59
- 239000000523 sample Substances 0.000 description 32
- 238000000034 method Methods 0.000 description 20
- 238000003384 imaging method Methods 0.000 description 16
- 238000002474 experimental method Methods 0.000 description 14
- 230000035772 mutation Effects 0.000 description 12
- 238000005516 engineering process Methods 0.000 description 7
- 238000012163 sequencing technique Methods 0.000 description 7
- 238000004590 computer program Methods 0.000 description 6
- 238000004364 calculation method Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 238000003364 immunohistochemistry Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 238000004891 communication Methods 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 238000012151 immunohistochemical method Methods 0.000 description 4
- 239000002207 metabolite Substances 0.000 description 4
- 238000001356 surgical procedure Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 238000001291 vacuum drying Methods 0.000 description 3
- 101001042041 Bos taurus Isocitrate dehydrogenase [NAD] subunit beta, mitochondrial Proteins 0.000 description 2
- 101000599885 Dictyostelium discoideum Isocitrate dehydrogenase [NADP], mitochondrial Proteins 0.000 description 2
- 101000960234 Homo sapiens Isocitrate dehydrogenase [NADP] cytoplasmic Proteins 0.000 description 2
- 102100039905 Isocitrate dehydrogenase [NADP] cytoplasmic Human genes 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000003759 clinical diagnosis Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000001869 matrix assisted laser desorption--ionisation mass spectrum Methods 0.000 description 2
- 238000001254 matrix assisted laser desorption--ionisation time-of-flight mass spectrum Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000010827 pathological analysis Methods 0.000 description 2
- 239000013610 patient sample Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000002271 resection Methods 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- MZNYWPRCVDMOJG-UHFFFAOYSA-N N-(1-naphthyl)ethylenediamine dihydrochloride Chemical compound [Cl-].[Cl-].C1=CC=C2C([NH2+]CC[NH3+])=CC=CC2=C1 MZNYWPRCVDMOJG-UHFFFAOYSA-N 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000004973 liquid crystal related substance Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a computing device, a storage medium, a glioma IDH typing detection device and a glioma IDH typing detection system. The computing device comprises a processor and a memory, wherein the memory stores programs, and the processor executes the programs to realize that: receiving tissue slice mass spectrum data of a glioma sample to be detected of a detected subject; obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity; and determining and judging the glioma IDH type of the examinee to be a mutant type or a wild type according to the comparison relation between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
Description
Technical Field
The invention relates to the field of medical clinical detection, in particular to a computing device, a storage medium and a glioma IDH typing detection device and system.
Background
In the early stages of glioma formation, there are two possible mutants of Isocitrate Dehydrogenase (IDH), IDH1 and IDH 2. Among them, IDH mutation is common in glioma cases, so detection of IDH mutation becomes one of important indexes for diagnosing glioma and medication guidance thereof.
The current clinical diagnosis for IDH mutations is mainly assessed by whole genome post-extraction sequencing and immunohistochemical methods of antibody capture against IDH muteins. In the standard process of clinical detection, the diagnosis is carried out by using an immunohistochemical method, the IDH mutation is confirmed if the result is positive, and the further sequencing judgment is carried out if the result is negative. Therefore, the method has certain difficulties in both the time consumption of experiments and the accuracy of diagnosis. In particular, immunohistochemistry and sequencing cannot assist pathological diagnosis in an operation due to time-consuming and complex experiments, so that obvious technical limitations exist in excision in the operation and timely medication guidance after the operation.
The MALDI-TOF mass spectrometry technology is determined to have accurate detection capability on the biological target markers in various tumors, and the development of the potential application of the MALDI-TOF mass spectrometry technology is also paid more and more attention in the tumor research field. In recent years, 2-HG (hydroxyglutarate) has also gained more and more attention as a bio-targeting marker for diagnosing glioma mutants. 2-HG is one of metabolites of glioma caused by IDH enzyme metabolism change generated by IDH mutation, and the accumulation of the 2-HG in glioma is proved to be accurately detected by MALDI-TOF mass spectrum and mass spectrum imaging technology thereof, thereby helping to diagnose the type of glioma IDH mutation.
Disclosure of Invention
An object of the present invention is to provide a computing device, a storage medium, and a glioma IDH typing detection apparatus and system.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention relates to a glioma IDH typing detection method, which comprises the following steps
Receiving tissue slice mass spectrum data of a glioma sample to be detected of a detected subject;
obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity;
and determining and judging the glioma IDH type of the examinee to be a mutant type or a wild type according to the comparison relation between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
Optionally, the determining that the glioma IDH of the subject is typed as a mutant type or a wild type according to the comparison relationship between the ratio of the hydroxyglutarate peak intensity to the sum of the hydroxyglutarate peak intensity, the glutamate peak intensity and the glutamine peak intensity and a preset typing value comprises:
if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is larger than a preset typing value, determining that the glioma IDH of the detected person is typed as a mutant type; and if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is less than a preset typing value, determining that the glioma IDH type of the detected person is a wild type.
Optionally, the preset typing value is 0.141069046504922.
Optionally, before receiving tissue section mass spectral data of a glioma sample to be detected of a subject, the method further comprises:
preparing a tissue section of a glioma sample to be detected of a detected subject;
carrying out matrix spraying covering on the tissue slice of the glioma sample to be detected of the detected object by using a spraying instrument;
and collecting the tissue section of the glioma sample to be detected after the matrix is sprayed by using a mass spectrometer to obtain the mass spectrum data of the tissue section of the glioma sample to be detected.
A second aspect of the invention relates to a computing device comprising a processor and a memory, the memory having stored therein a program that when executed by the processor effects:
receiving tissue slice mass spectrum data of a glioma sample to be detected of a detected subject;
obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity;
and determining and judging the glioma IDH type of the examinee to be a mutant type or a wild type according to the comparison relation between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
Optionally, the determining that the glioma IDH of the subject is typed as a mutant type or a wild type according to the comparison relationship between the ratio of the hydroxyglutarate peak intensity to the sum of the hydroxyglutarate peak intensity, the glutamate peak intensity and the glutamine peak intensity and a preset typing value comprises:
if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is larger than a preset typing value, determining that the glioma IDH of the detected person is typed as a mutant type; and if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is less than a preset typing value, determining that the glioma IDH type of the detected person is a wild type.
Optionally, the preset typing value is 0.141069046504922.
A third aspect of the present invention relates to a storage medium storing a program that, when executed, realizes:
receiving tissue slice mass spectrum data of a glioma sample to be detected of a detected subject;
obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity;
and determining and judging the glioma IDH type of the examinee to be a mutant type or a wild type according to the comparison relation between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
Optionally, the determining that the glioma IDH of the subject is typed as a mutant type or a wild type according to the comparison relationship between the ratio of the hydroxyglutarate peak intensity to the sum of the hydroxyglutarate peak intensity, the glutamate peak intensity and the glutamine peak intensity and a preset typing value comprises:
if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is larger than a preset typing value, determining that the glioma IDH of the detected person is typed as a mutant type; and if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is less than a preset typing value, determining that the glioma IDH type of the detected person is a wild type.
Optionally, the preset typing value is 0.141069046504922.
A fourth aspect of the present invention relates to a glioma IDH typing detection device, comprising:
the spraying instrument is used for carrying out matrix spraying and covering on the tissue section of the glioma sample to be detected of the detected person;
the mass spectrometer is used for collecting the tissue section of the glioma sample to be detected after the matrix is sprayed, so as to obtain the mass spectrum data of the tissue section of the glioma sample to be detected; and
the computing device of the second aspect of the invention.
The fifth aspect of the invention relates to a glioma IDH typing detection system, which comprises
The apparatus of the fourth aspect of the invention;
and the display is used for displaying the judgment result.
Optionally, the spray applicator is an HTX spray applicator.
Optionally, the mass spectrometer is a QuanTOF II MALDI-TOF mass spectrometer.
The invention has the following beneficial effects:
the method comprises the steps of spraying a mixed matrix on tissue slices of a glioma sample to be detected by using a spraying instrument, collecting mass spectrum data of the tissue slices of the glioma sample to be detected by using a mass spectrometer, calculating the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram result by combining software, and successfully and accurately distinguishing the IDH mutant type and the wild type by comparing the ratio with a preset typing value. Therefore, the method is superior to the second Generation gene Sequencing (NGS) and Immunohistochemistry (IHC) detection in the prior art in terms of long detection time, personnel consumption, material and cost, the total experiment time can be controlled within one hour to give an experiment conclusion, the pathological IDH typing of the tissue section is judged, each experiment has simple experiment step operation, and the acquisition speed and analysis can achieve full automation. In addition, only ITO slides, chemical substrates and isotope internal standards after dilution ratio are consumed by the materials, so that the cost of the experiment is greatly saved; the sample slice does not need to consume a large amount of tissues, and only a single sample with the thickness of 10-12 microns is needed, so that the current molecular distribution intensity information of the complete slice can be given. All the interpretation results have complete spectrogram data and imaging data tracing with high typing interpretation accuracy, and the repeated acquisition is supported by the same method, so that the repeated verification of the data can be carried out.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
Fig. 1 shows a flowchart of a glioma IDH typing detection method provided by the embodiment of the present invention.
Fig. 2 shows a schematic diagram of one example of imaging data from 111 real patient samples.
FIG. 3 shows a graphical plot of peak intensity for hydroxyglutaric acid (2-HG), glutamic acid (Glu), and glutamine (Gln) versus the imaging data of FIG. 2.
Fig. 4 shows a schematic representation of imaging data of yet another of 111 real patient samples.
FIG. 5 shows a graphical plot of peak intensity corresponding to 2-HG, Glu, and Gln of the imaging data of FIG. 4.
FIG. 6 is a graph showing CV check results of the ratio calculation of 111 real patients as independent samples.
FIG. 7 is a schematic diagram of ROC for ratio calculation using the detection method provided by the present invention.
FIG. 8 illustrates a computer system architecture diagram suitable for implementing the detection method of an embodiment of the present invention.
Detailed Description
In order to more clearly illustrate the invention, the invention is further described below with reference to preferred embodiments and the accompanying drawings. Similar parts in the figures are denoted by the same reference numerals. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.
In the early stages of glioma formation, there are two possible mutants of Isocitrate Dehydrogenase (IDH), IDH1 and IDH 2. Among them, mutations of IDH are common in cases of glioma, and thus detection of IDH mutations is a common means for diagnosing glioma. The current clinical diagnosis for IDH mutations is mainly assessed by whole genome post-extraction sequencing and immunohistochemical methods of antibody capture against IDH muteins. In the standard process of clinical detection, the diagnosis is carried out by using an immunohistochemical method, the IDH mutation is confirmed if the result is positive, and the further sequencing judgment is carried out if the result is negative. Therefore, the method has certain difficulties in both the time consumption of experiments and the accuracy of diagnosis. In particular, immunohistochemistry and sequencing cannot assist pathological diagnosis in surgery due to time-consuming and complicated experiments, and thus have obvious limitations in resection in surgery and timely medication guidance after surgery.
The MALDI-TOF mass spectrometry technology is determined to have accurate detection capability on the biological target markers in various tumors, and the development of the potential application of the MALDI-TOF mass spectrometry technology is also paid more and more attention in the tumor research field. In recent years, 2-HG (hydroxyglutarate) has also gained more and more attention as a bio-targeting marker for diagnosing glioma mutants. 2-HG is one of metabolites of glioma caused by IDH enzyme metabolism change generated by IDH mutation, and the accumulation of the 2-HG in glioma is proved to be accurately detected by MALDI-TOF mass spectrum and mass spectrum imaging technology thereof, thereby helping to diagnose the type of glioma IDH mutation.
The inventor finds that the glioma IDH typing qualitative proportion operation is carried out according to the target small molecule metabolite, such as 2-HG, and the amino acids with similar mass, so that the IDH mutant type and the wild type can be distinguished efficiently and accurately.
Thus, as shown in fig. 1, an embodiment of the present invention provides a glioma IDH typing detection method, including:
s101, receiving mass spectrum data of a tissue slice of a glioma sample to be detected of a detected person;
s103, obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in the mass spectrogram result of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity;
s105, determining and judging whether the glioma IDH of the detected person is of a mutant type or a wild type according to the comparison relationship between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
Optionally, the determining that the glioma IDH of the subject is typed as a mutant type or a wild type according to the comparison relationship between the ratio of the hydroxyglutarate peak intensity to the sum of the hydroxyglutarate peak intensity, the glutamate peak intensity and the glutamine peak intensity and a preset typing value comprises:
if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is larger than a preset typing value, determining that the glioma IDH of the detected person is typed as a mutant type; and if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is less than a preset typing value, determining that the glioma IDH type of the detected person is a wild type.
Optionally, the preset typing value is 0.141069046504922.
Optionally, before receiving tissue section mass spectral data of a glioma sample to be detected of a subject, the method further comprises:
preparing a tissue section of a glioma sample to be detected of a detected subject;
carrying out matrix spraying covering on the tissue slice of the glioma sample to be detected of the detected object by using a spraying instrument;
and collecting the tissue section of the glioma sample to be detected after the matrix is sprayed by using a mass spectrometer to obtain the mass spectrum data of the tissue section of the glioma sample to be detected.
In a specific example, a glioma sample to be detected of the subject of the present invention is obtained from an operation, and the specific operation flow of the glioma IDH typing detection method provided by the embodiment of the present invention is as follows:
(1) obtaining materials in the operation: excising a glioma sample in a clinical glioma resection procedure;
(2) freezing and slicing: the glioma specimens cut out in the surgery are subjected to liquid nitrogen flash freezing, after which frozen sections of the tissue are taken using a cryomicrotome, for example a leica cryomicrotome, with a section thickness of 10 to 12 μm. The slicing temperature in the freezing slicer is-20 ℃ to-22 ℃, the slicing is recommended to be the middle position of the tissue, the comprehensiveness of the tissue is ensured as far as possible, and the cut tissue is attached to an ITO glass slide;
(3) and (3) vacuum drying: carrying out rapid vacuum drying on the ITO glass slide attached with the tissue for 5 minutes by using a vacuum drying box, wherein the pressure is more than 15 psi;
(4) matrix spraying:
preparing a matrix: NEDC (N- (1-naphthyl) ethylenediamine dihydrochloride) was placed in methanol: in a solution containing 70% water and 30% (v: v), NEDC was prepared at a ratio of 10mg/mL, and an internal standard was used13C5Isotopically labelled 2-HG, added to a prepared matrix, the matrix13C5-concentration of 2-HG is 500 pmol/. mu.L;
spraying by a spraying instrument: and (3) carrying out matrix spraying covering on the dried ITO glass slide attached with the tissue by using a spraying instrument, wherein the spraying instrument adopts a spraying instrument HTX TM-spraying (HTX Imaging technologies) mass spectrometer, and the experimental parameters are set as follows: the liquid flow rate is 0.07mL/min, the spraying layer number (passes) is 10, the spraying temperature is 75 ℃, and the nitrogen pressure is 10 psi; the drying time is 3 seconds, and the moving speed is 1000 mm/min;
(5) collecting by a mass spectrometer: collecting the tissue slice of the glioma sample to be detected after matrix spraying by using a mass spectrometer to obtain the mass spectrum data of the tissue slice of the glioma sample to be detected, wherein the mass spectrometer is a QuandTOF II MALDI-TOF mass spectrometer, and the parameters are set as follows: an acquisition mode: linear Negative Ion Mode (Linear Negative Ion Mode), spatial resolution: 50 μm2The focusing quality is as follows: 140m/z, laser frequency: 5,000Hz, mass range: 50-1,000m/z, acceleration voltage: 10kV, detection voltage: -0.8kV, laser energy: 12 μ J, laser diameter: 10 μm, number of single spectrum laser bombardment: 50 shotts;
(6) the computing device analyzes:
spectrogram adjustment and imaging:
opening the acquired mass spectrum data by using analysis software QuanViewer of computing equipment, performing data baseline calibration and internal standard mass calibration, performing one-click imaging, selecting an area in a tissue slice, performing ratio calculation result checking of the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity, and comparing the established standard typing cut-off value (namely a preset typing value) to obtain a glioma IDH typing result, namely a mutant type or a wild type, of the subject based on relative quantification.
In a specific exampleIn the middle, the accuracy of baseline correction is 10, and after automatic internal calibration, the internal standard13C5Imaging quality of-2-HG (M-H)+) The imaging mass (M-H) of the target metabolite hydroxyglutaric acid 2-HG is obtained with the internal standard as mass calibration at 151.35M/z+) 147.00M/z, error between plus and minus 0.5Da, imaging Mass of Glutamine Gln (M-H)+) 145.15M/z, error between plus and minus 0.5Da, imaging mass (M-H) of glutamic acid Glu+) 146.13m/z, with an error of between plus or minus 0.5 Da.
Proportional operation:
and (4) carrying out proportional operation on the peak intensity: comparing the ratio of the peak intensity of 2-HG/(2-HG + Gln + Glu) with the cut-off value to determine the IDH type, and if the ratio of the peak intensity of 2-HG/(2-HG + Gln + Glu) is greater than the cut-off value, determining that the result is mutant; and if the ratio of the peak intensity of the 2-HG/(2-HG + Gln + Glu) to the peak intensity is less than the cut-off value, the result is the wild type, wherein the cut-off value is calculated based on hundreds of real independent glioma samples and the calculation of the ROC curve and KS statistics.
Next, the statistical law is verified.
In one specific example, NGS is the gold standard for clinical molecular diagnostics of the current IDH typing, so the inventors compared the results obtained by the detection method provided by the present invention with the results of the existing NGS method for 111 independent samples (from 42 real patients) all having NGS results.
2-HG, Glu and Gln can be detected from 111 real patients by using the detection method provided by the invention, and as shown in fig. 2-5, 2 imaging data of 111 real patients and peak intensity maps corresponding to 2-HG, Gln and Glu are obtained.
The section data of 111 real patients are respectively regarded as independent samples to be subjected to CV check based on ratio calculation, as shown in FIG. 6, the probability that the finished data CV is greater than 0.1 and is 9.17 percent is obtained, namely, the detection method provided by the invention has low operation dispersion degree.
And calculating the consistency by combining all the results, wherein the kappa is 0.92695652, which shows that the detection method provided by the invention has higher consistency with the NGS method.
And analyzing the ratio of the hydroxyglutarate peak intensity to the sum of the hydroxyglutarate peak intensity, the glutamic acid peak intensity and the glutamine peak intensity by combining all samples to obtain a clear cut-off value limit between the mutant type and the wild type, wherein as shown in fig. 7, the ROC result can determine the significant relation between the mutant type and the wild type to obtain AUC (0.954), CI (95%) and cut-off (0.141069046504922).
The method comprises the steps of spraying a mixed matrix on tissue slices of a glioma sample to be detected by using a spraying instrument, collecting mass spectrum data of the tissue slices of the glioma sample to be detected by using a mass spectrometer, calculating the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram result by combining software, and successfully and accurately distinguishing the IDH mutant type and the wild type by comparing the ratio with a preset typing value. Therefore, the invention realizes the NGS and IHC detection superior to the prior art in multiple angles of long detection time, personnel consumption, material and cost. The whole experiment duration can be controlled within one hour to give an experiment conclusion, the tissue section pathology IDH typing is judged, each step of experiment has simple and mature experiment step operation, and the acquisition and analysis can be fully automatic. In addition, only ITO slides, chemical substrates and isotope internal standards after dilution ratio are consumed by the materials, so that the cost of the experiment is greatly saved; the sample slice does not need to consume a large amount of tissues, and only a single sample with the thickness of 10-12 microns is needed, so that the current molecular distribution intensity information of the complete slice can be given. All the interpretation results have complete spectrogram data and imaging data tracing with high typing interpretation accuracy, and the repeated acquisition is supported by the same method, so that the repeated verification of the data can be carried out.
The invention provides a computing device comprising a processor which, when executing a program, carries out the method steps shown in figure 1.
In a specific example, as shown in fig. 8, a computer system suitable for implementing the glioma IDH typing detection method provided by the present embodiment includes a Central Processing Unit (CPU), which can execute the method according to a program stored in a Read Only Memory (ROM) or a program loaded from a storage portion into a Random Access Memory (RAM), for example, an application program implementing the recruitment method of the present application or when executing the program. The CPU, ROM, and RAM are connected thereto via a bus. An input/output (I/O) interface is also connected to the bus.
An input section including a keyboard, a mouse, and the like; an output section including a speaker and the like such as a Liquid Crystal Display (LCD); a storage section including a hard disk and the like; and a communication section including a network interface card such as a LAN card, a modem, or the like. The communication section performs communication processing via a network such as the internet. The drive is also connected to the I/O interface as needed. A removable medium such as a magnetic disk, an optical disk, a magneto-optical disk, a semiconductor memory, or the like is mounted on the drive as necessary, so that a computer program read out therefrom is mounted into the storage section as necessary.
In particular, the processes described in the above flowcharts may be implemented as computer software programs according to the present embodiment. For example, the present embodiments include a computer program product comprising a computer program tangibly embodied on a computer-readable medium, the computer program comprising program code for performing the method illustrated in the flow chart. In such an embodiment, the computer program may be downloaded and installed from a network via the communication section, and/or installed from a removable medium.
The flowchart and schematic diagrams in the figures illustrate the architecture, functionality, and operation of possible implementations of systems, methods and computer program products according to the present embodiments. In this regard, each block in the flowchart or schematic diagrams may represent a module, segment, or portion of code, which comprises one or more executable instructions for implementing the specified logical function(s). It should also be noted that, in some alternative implementations, the functions noted in the block may occur out of the order noted in the figures. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocks may sometimes be executed in the reverse order, depending upon the functionality involved. It will also be noted that each block of the schematic and/or flowchart illustration, and combinations of blocks in the schematic and/or flowchart illustration, can be implemented by special purpose hardware-based systems which perform the specified functions or acts, or combinations of special purpose hardware and computer instructions.
The modules referred to in the embodiments described may be implemented by means of software. The modules described may also be provided in a processor.
On the other hand, the present embodiment also provides a nonvolatile computer storage medium, which may be the nonvolatile computer storage medium included in the apparatus in the foregoing embodiment, or may be a nonvolatile computer storage medium that exists separately and is not assembled into a terminal. The non-transitory computer storage medium stores one or more programs that, when executed by a device, cause the device to perform the glioma IDH typing detection method of the present application.
The invention also provides a glioma IDH typing detection device, which comprises
The spraying instrument is used for carrying out matrix spraying and covering on the tissue section of the glioma sample to be detected of the detected person;
the mass spectrometer is used for collecting the tissue section of the glioma sample to be detected after the matrix is sprayed, so as to obtain the mass spectrum data of the tissue section of the glioma sample to be detected; and the computing device described above.
The invention also provides a glioma IDH typing detection system, which comprises
The glioma IDH typing detection device; and
and the display is used for displaying the judgment result.
Specifically, the result report is shown, for example, as "mutant type" and "wild type".
Optionally, the spray applicator is an HTX spray applicator.
Optionally, the mass spectrometer is a QuanTOF II MALDI-TOF mass spectrometer.
It should be understood that the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention, and it will be obvious to those skilled in the art that other variations or modifications may be made on the basis of the above description, and all embodiments may not be exhaustive, and all obvious variations or modifications may be included within the scope of the present invention.
Claims (10)
1. A computing device comprising a processor and a memory, the memory having a program stored therein, the processor when executing the program implementing:
receiving tissue slice mass spectrum data of a glioma sample to be detected of a detected subject;
obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity;
and determining and judging the glioma IDH type of the examinee to be a mutant type or a wild type according to the comparison relation between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
2. The computing device of claim 1, wherein determining whether the subject's glioma IDH is typed as mutant or wild type based on a comparison of the ratio of the hydroxyglutarate peak intensity to the sum of hydroxyglutarate peak intensity, glutamate peak intensity, and glutamine peak intensity to a preset typing value comprises:
if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is larger than a preset typing value, determining that the glioma IDH of the detected person is typed as a mutant type; and if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is less than a preset typing value, determining that the glioma IDH type of the detected person is a wild type.
3. The computing device of claim 2,
the preset typing value is 0.141069046504922.
4. A storage medium storing a program, the program when executed implementing:
receiving tissue slice mass spectrum data of a glioma sample to be detected of a detected subject;
obtaining the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity in a mass spectrogram of the tissue slice based on mass spectrum data, and calculating the ratio of the hydroxyglutaric acid peak intensity to the sum of the hydroxyglutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity;
and determining and judging the glioma IDH type of the examinee to be a mutant type or a wild type according to the comparison relation between the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity and a preset typing value.
5. The storage medium of claim 4, wherein the determining whether the subject's glioma IDH is typed as mutant or wild type based on the comparison of the ratio of the hydroxyglutarate peak intensity to the sum of the hydroxyglutarate peak intensity, the glutamate peak intensity, and the glutamine peak intensity to a preset typing value comprises:
if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is larger than a preset typing value, determining that the glioma IDH of the detected person is typed as a mutant type; and if the ratio of the hydroxyl glutaric acid peak intensity to the sum of the hydroxyl glutaric acid peak intensity, the glutamic acid peak intensity and the glutamine peak intensity is less than a preset typing value, determining that the glioma IDH type of the detected person is a wild type.
6. The storage medium of claim 5,
the preset typing value is 0.141069046504922.
7. A glioma IDH typing detection device, comprising:
the spraying instrument is used for carrying out matrix spraying and covering on the tissue section of the glioma sample to be detected of the detected person;
the mass spectrometer is used for collecting the tissue section of the glioma sample to be detected after the matrix is sprayed, so as to obtain the mass spectrum data of the tissue section of the glioma sample to be detected; and
the computing device of any of claims 1-3.
8. A glioma IDH typing detection system is characterized by comprising
The device of claim 7;
and the display is used for displaying the judgment result.
9. The system of claim 8, wherein the spray applicator is an HTX spray applicator.
10. The system of claim 8, wherein the mass spectrometer is a quanattof II MALDI-TOF mass spectrometer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111586578.6A CN114295836A (en) | 2021-12-22 | 2021-12-22 | Computing equipment, storage medium and glioma IDH typing detection device and system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111586578.6A CN114295836A (en) | 2021-12-22 | 2021-12-22 | Computing equipment, storage medium and glioma IDH typing detection device and system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114295836A true CN114295836A (en) | 2022-04-08 |
Family
ID=80968733
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111586578.6A Pending CN114295836A (en) | 2021-12-22 | 2021-12-22 | Computing equipment, storage medium and glioma IDH typing detection device and system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114295836A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106999490A (en) * | 2014-08-07 | 2017-08-01 | 卡利泰拉生物科技公司 | The glutamine enzyme inhibitor of crystal form |
| CN110072550A (en) * | 2016-11-01 | 2019-07-30 | 德那翠丝有限公司 | For treating the combination treatment of the cancer of the brain |
| CN113588768A (en) * | 2021-05-18 | 2021-11-02 | 国家卫生健康委科学技术研究所 | Mass spectrometry method for quantifying endogenous metabolites in tissues in molecular image mode |
-
2021
- 2021-12-22 CN CN202111586578.6A patent/CN114295836A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106999490A (en) * | 2014-08-07 | 2017-08-01 | 卡利泰拉生物科技公司 | The glutamine enzyme inhibitor of crystal form |
| CN110072550A (en) * | 2016-11-01 | 2019-07-30 | 德那翠丝有限公司 | For treating the combination treatment of the cancer of the brain |
| CN113588768A (en) * | 2021-05-18 | 2021-11-02 | 国家卫生健康委科学技术研究所 | Mass spectrometry method for quantifying endogenous metabolites in tissues in molecular image mode |
Non-Patent Citations (1)
| Title |
|---|
| JUDITH M. KAMPA等: "Glioblastoma multiforme: Metabolic differences to peritumoral tissue and IDH-mutated gliomas revealed by mass spectrometry imaging", NEUROPATHOLOGY, 13 July 2020 (2020-07-13), pages 546 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110243921B (en) | Rapid tumor tissue discrimination method based on tissue surface lipid fingerprint spectrogram | |
| Coombes et al. | Quality control and peak finding for proteomics data collected from nipple aspirate fluid by surface-enhanced laser desorption and ionization | |
| Seeley et al. | Molecular imaging of proteins in tissues by mass spectrometry | |
| Yalcin et al. | Review of matrix-assisted laser desorption ionization-imaging mass spectrometry for lipid biochemical histopathology | |
| Balluff et al. | Batch effects in MALDI mass spectrometry imaging | |
| Ryan et al. | Protein identification in imaging mass spectrometry through spatially targeted liquid micro‐extractions | |
| CN114414704B (en) | System, model and kit for evaluating malignancy degree or probability of thyroid nodule | |
| Ye et al. | A vision for better health: mass spectrometry imaging for clinical diagnostics | |
| CN109557165B (en) | Method for monitoring the quality of a mass spectrometry imaging preparation workflow | |
| US10197576B2 (en) | Mass spectrometry imaging with substance identification | |
| US20180088094A1 (en) | Multiple attribute monitoring methodologies for complex samples | |
| US20040172200A1 (en) | Constellation mapping and uses thereof | |
| Janda et al. | Determination of abundant metabolite matrix adducts illuminates the dark metabolome of MALDI-mass spectrometry imaging datasets | |
| Minerva et al. | MALDI MS imaging as a tool for biomarker discovery: methodological challenges in a clinical setting | |
| Chung et al. | Next‐generation pathology practices with mass spectrometry imaging | |
| Djambazova et al. | Advances in imaging mass spectrometry for biomedical and clinical research | |
| GB2607200A (en) | Mass spectrometric determination of particular tissue states | |
| JP6731957B2 (en) | Method of diagnosing endometrial cancer | |
| CN114295836A (en) | Computing equipment, storage medium and glioma IDH typing detection device and system | |
| EP3936870B1 (en) | Method for quantitation of her2 in breast cancer sample by mass spectrometry and scoring of her2 state by using same | |
| JP7158642B2 (en) | Mass spectrometer and mass spectrometry system | |
| EP1587406B1 (en) | Quantification of carnitine levels in dialysis patients | |
| EP3924730B1 (en) | Apparatus and method for targeted compound analysis | |
| CN117849159A (en) | M protein detection method, electronic equipment and storage medium | |
| Cabello-Pinedo et al. | Development of a Novel Non-invasive Metabolomics Assay to Predict Implantation Potential of Human Embryos |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |