CN117106876A - Method for detecting gonomic cancers based on high-throughput sequencing research and development - Google Patents
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Abstract
The invention discloses a detection method of stranguria blood cancer based on high-flux sequencing research and development, and relates to the technical field of high-flux sequencing. The detection method of the gonococcal hematologic cancer based on high-throughput sequencing development comprises the following steps: step one: sampling, namely slicing and sampling the enlarged lymph nodes of a patient by using a cutting scalpel, and simultaneously placing the initial sample into physiological saline for storing the initial sample. According to the method for detecting the stranguria blood cancer based on high-throughput sequencing research and development, the detection times of a single sample can be improved by diluting the sample, so that the detection precision of the sample can be improved, the repeated sampling caused by detection errors is avoided, the detection efficiency is improved, meanwhile, different treatment schemes are designated for patients with different conditions by grading the samples according to the ratio of cancer cells, the utilization rate of medical resources is improved, and the medical resources are prevented from being wasted.
Description
Technical Field
The invention relates to the technical field of high-throughput sequencing, in particular to a method for detecting stranguria blood cancer based on high-throughput sequencing research and development.
Background
With the development of society and the improvement of living standard, people also pay more attention to their physical health. However, many diseases still threaten human health, affect human life quality, such as cancer, and although medical staff can judge cancer from historical diagnosis information, it is necessary to process a large amount of information and data sets, so how to effectively and correctly detect cancer and perform relevant treatment becomes an important technology. Also, since a large amount of medical data information exists in reality, it is very important how to select important data information from the medical data information to detect cancer. Therefore, it is necessary to remove noise information and redundant information from a large amount of medical data, and screen out important sample learning models for effective detection.
The invention patent with the patent publication number of CN109360656B discloses a cancer detection method based on a multi-target evolution algorithm, which comprises the following steps: screening an initial cancer data set through a multi-target evolutionary algorithm to obtain a cancer detection data model; the accuracy of the cancer detection data model is improved through a multi-target integration algorithm, and the cancer detection integrated data model is obtained; cancer detection is performed on the target cancer dataset through the cancer detection integrated data model. Therefore, the decompression rate of the first front surface is greatly improved, the classification accuracy is improved, the performance of the algorithm is improved by adopting multi-target integration, the number of the selected integrated models is greatly reduced, and the classification accuracy of the algorithm on cancer samples is further improved by the multi-target integration.
Although the above patent can reduce the number of selected integrated models, and further improve the accuracy of classifying cancer samples by the algorithm through multi-objective integration, in a specific detection process, high throughput sequencing is generally cited to improve the measurement efficiency, but the existing measurement reports are provided for the samples, if detection errors or sample pollution occur in the detection process, sampling operation is required to be performed again, and meanwhile, a recommended treatment means cannot be provided directly in the detection process, so that the detection effect is poor.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for detecting the stranguria blood cancer based on high-flux sequencing research and development, which solves the problems in the background art.
In order to achieve the above purpose, the invention is realized by the following technical scheme: a detection method of gonococcal blood cancer developed based on high-throughput sequencing comprises the following steps:
step one: sampling, namely slicing and sampling the enlarged lymph node of a patient by using a cutting scalpel, and simultaneously placing the initial sample into physiological saline for storing the initial sample;
step two: diluting the sample, namely diluting the initial sample in the normal saline in the first step in diluent, and cutting the initial sample by using a fine cutter under an electron microscope to obtain diluted samples ten times as many as the initial sample;
step three: sample storage, namely placing the diluted sample obtained in the second step into a refrigeration device for storage, and controlling the temperature of the refrigeration device to be the temperature for inhibiting cell division;
step four: sample detection, namely sequentially detecting ten diluted samples stored in the step three, wherein the specific detection process is as follows:
s1: immobilizing the amplified microspheres on a glass substrate to form a high throughput array;
s2: hybridizing the universal primer with a DNA library template attached to the microsphere, and then performing a series of ligation reactions, each ligation reaction occurring between the DNA extension strand and a probe in a pool of fluorescently labeled single-stranded octanucleotide probes;
s3: the base of the octanucleotide probe has a definite corresponding relation with a specific fluorescent color, after a series of complex connection, enzyme digestion and reaction cycle of the combination of the next primer, a fluorescent image is obtained, and DNA sequence information can be read out according to the corresponding relation between the base and the fluorescence, so that the number of cancer cells is obtained;
step five: counting the number of the cancer cells detected in each diluted sample in the fourth step, and recording the number of the cancer cells as C SFC And counting the specific number of cells in each sample under the electron microscope in the second step in advance, and simultaneously recording the specific number of cells in each sample as K SFC ；
Step six: for a pair ofStep five, counting the number of cancer cells and all cells obtained in step seven by referring to the following formula: for the cancer cell fraction P obtained in the step six SFC Rating was performed with the following criteria: and (3) calculating: p (P) SFC =C SFC /K SFC ，（P SFC Is the ratio of cancer cells); and (3) calculating: p (P) SFC =C SFC /K SFC ，（P SFC Is the ratio of cancer cells);
step seven: for the cancer cell fraction P obtained in the step six SFC Rating was performed with the following criteria:
first stage: p (P) SFC If the ratio is more than or equal to 0.6 and less than 1, the cancer of cells in the sample is serious, and strong treatment is needed in a targeted manner;
second stage: p (P) SFC If the ratio is more than or equal to 0.4 and less than 0.6, the canceration of cells in the sample is heavier, and strong treatment is needed selectively;
third stage: p (P) SFC If the ratio is more than or equal to 0.2 and less than 0.4, the canceration intensity of cells in the sample is medium, and medium-intensity treatment is needed to be selectively carried out;
fourth stage: p (P) SFC More than or equal to 0.01 and less than 0.2, the cancer strength of cells in the sample is weak, and chemotherapy and medication are selected in a targeted manner;
step eight: the treatment scheme is formulated according to the stranguria blood cancer grade defined in the step seven to carry out targeted treatment, the detection times of a single sample can be improved by diluting the sample, so that the detection precision of the sample can be improved, the repeated sampling caused by detection errors is avoided, the detection efficiency is improved, and meanwhile, the grading of each sample according to the ratio of cancer cells is beneficial to designating different treatment schemes for patients with different conditions, so that the utilization rate of medical resources is improved, and the medical resources are prevented from being wasted.
Preferably, the physiological saline in the first step is 0.9% sodium chloride solution, and the temperature of the physiological saline is 35.5-37.2 ℃.
Preferably, the main components of the diluent in the second step are glacial acetic acid and methylene blue, and the total amount of the diluent is ten times that of the physiological saline in the first step.
Preferably, in the step two, in the process of cutting the initial sample, a fine cutter is used to remove the redundant tissue in the initial sample.
Preferably, in the sample storage process in the third step, the storage time of the diluted sample is recorded, the content of each cell in the diluted sample is correspondingly reduced according to the division speed of cancer cells and common cells, the time difference between rapid sampling and slow detection can be balanced through the storage of the samples in a low-temperature environment, so that the taken sample can be well stored, the serious error of the sample in the subsequent detection caused by rapid splitting of the taken sample is prevented, meanwhile, the large deviation in calculation of the number of fine cutters and the cells can be avoided by utilizing an electron microscope, and the data accuracy is improved.
Preferably, the glass substrate in the fourth step includes a plurality of separation areas, and no more than two diluted samples are disposed in each separation area, so as to avoid mutual pollution between the diluted samples and influence on the detection result.
Preferably, the cancer cell fraction rating in the step seven is derived in the form of a written report or an electronic report, and the treatment instruction advice is attached with reference to the rating content definition.
Preferably, the patients rated in the seventh step are subjected to sub-sampling detection with the first and fourth grades to ensure data accuracy.
The invention provides a method for detecting stranguria blood cancer based on high-throughput sequencing research and development. The beneficial effects are as follows:
according to the method for detecting the stranguria blood cancer based on high-throughput sequencing research and development, the detection times of a single sample can be improved by diluting the sample, so that the detection precision of the sample can be improved, the repeated sampling caused by detection errors is avoided, the detection efficiency is improved, meanwhile, different treatment schemes are designated for patients with different conditions by grading the samples according to the ratio of cancer cells, the utilization rate of medical resources is improved, and the medical resources are prevented from being wasted.
According to the method for detecting the stranguria blood cancer based on high-throughput sequencing research and development, the time difference between rapid sampling and slow detection can be balanced through storage of samples in a low-temperature environment, so that the taken samples can be stored well, serious errors of the samples in subsequent detection caused by rapid splitting of the taken samples are prevented, meanwhile, large deviation of fine cutters and cell number calculation can be avoided by utilizing an electron microscope, and data accuracy is improved.
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FIG. 1 is a block diagram of the overall structure of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1, the present invention provides a technical solution: a detection method of gonococcal blood cancer developed based on high-throughput sequencing comprises the following steps:
step one: sampling, namely slicing and sampling the enlarged lymph node of a patient by using a cutting scalpel, and simultaneously placing the initial sample into physiological saline for storing the initial sample;
step two: diluting the sample, namely diluting the initial sample in the normal saline in the first step in diluent, and cutting the initial sample by using a fine cutter under an electron microscope to obtain diluted samples ten times as many as the initial sample;
step three: sample storage, namely placing the diluted sample obtained in the second step into a refrigeration device for storage, and controlling the temperature of the refrigeration device to be the temperature for inhibiting cell division;
step four: sample detection, namely sequentially detecting ten diluted samples stored in the step three, wherein the specific detection process is as follows:
s1: immobilizing the amplified microspheres on a glass substrate to form a high throughput array;
s2: hybridizing the universal primer with a DNA library template attached to the microsphere, and then performing a series of ligation reactions, each ligation reaction occurring between the DNA extension strand and a probe in a pool of fluorescently labeled single-stranded octanucleotide probes;
s3: the base of the octanucleotide probe has a definite corresponding relation with a specific fluorescent color, after a series of complex connection, enzyme digestion and reaction cycle of the combination of the next primer, a fluorescent image is obtained, and DNA sequence information can be read out according to the corresponding relation between the base and the fluorescence, so that the number of cancer cells is obtained;
step five: counting the number of the cancer cells detected in each diluted sample in the fourth step, and recording the number of the cancer cells as C SFC And counting the specific number of cells in each sample under the electron microscope in the second step in advance, and simultaneously recording the specific number of cells in each sample as K SFC ；
Step six: the number of cancer cells and all cells obtained in the fifth step were calculated with reference to the following formula: p (P) SFC =C SFC /K SFC ，（P SFC Is the ratio of cancer cells);
step seven: for the cancer cell fraction P obtained in the step six SFC Rating was performed with the following criteria:
first stage: p (P) SFC If the ratio is more than or equal to 0.6 and less than 1, the cancer of cells in the sample is serious, and strong treatment is needed in a targeted manner;
second stage: p (P) SFC If the ratio is more than or equal to 0.4 and less than 0.6, the canceration of cells in the sample is heavier, and strong treatment is needed selectively;
third stage: p (P) SFC If the ratio is more than or equal to 0.2 and less than 0.4, the canceration intensity of cells in the sample is medium, and medium-intensity treatment is needed to be selectively carried out;
fourth stage: p (P) SFC More than or equal to 0.01 and less than 0.2, the cancer strength of cells in the sample is weak, and chemotherapy and medication are selected in a targeted manner;
step eight: the treatment scheme is formulated according to the stranguria blood cancer grade defined in the step seven to carry out targeted treatment, the detection times of a single sample can be improved by diluting the sample, so that the detection precision of the sample can be improved, the repeated sampling caused by detection errors is avoided, the detection efficiency is improved, and meanwhile, the grading of each sample according to the ratio of cancer cells is beneficial to designating different treatment schemes for patients with different conditions, so that the utilization rate of medical resources is improved, and the medical resources are prevented from being wasted.
The physiological saline in the first step is 0.9% concentration sodium chloride solution, and the temperature of the physiological saline is 35.5-37.2 ℃.
The main components of the diluent in the second step are glacial acetic acid and methylene blue, and the total amount of the diluent is ten times of that of the physiological saline in the first step.
In the process of cutting the initial sample, removing redundant tissues in the initial sample by using a fine cutter.
In the sample storage process in the third step, the time for diluting the sample is recorded, the content of each cell in the diluted sample is correspondingly reduced according to the division speed of cancer cells and common cells, the time difference between rapid sampling and slow detection can be balanced through the storage of the samples in a low-temperature environment, so that the taken sample can be well stored, the serious error of the sample in the subsequent detection caused by rapid division of the taken sample is prevented, meanwhile, the large deviation of fine cutters and cell number calculation can be avoided by utilizing an electron microscope, and the data accuracy is improved.
The glass substrate in the fourth step comprises a plurality of separation areas, and no more than two diluted samples are arranged in each separation area, so that the influence of mutual pollution among the diluted samples on the detection result is avoided.
The cancer cell fraction rating in the seventh step is derived in the form of a written report or an electronic report, and the treatment instruction advice is attached with reference to the rating content definition.
And D, performing subsampling detection on the patients with the rated grades of the first grade and the fourth grade in the step seven so as to ensure the accuracy of data.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. A detection method of gonococcal blood cancer developed based on high-throughput sequencing is characterized by comprising the following steps: the method comprises the following steps:
step one: sampling, namely slicing and sampling the enlarged lymph node of a patient by using a cutting scalpel, and simultaneously placing the initial sample into physiological saline for storing the initial sample;
step two: diluting the sample, namely diluting the initial sample in the normal saline in the first step in diluent, and cutting the initial sample by using a fine cutter under an electron microscope to obtain diluted samples ten times as many as the initial sample;
step three: sample storage, namely placing the diluted sample obtained in the second step into a refrigeration device for storage, and controlling the temperature of the refrigeration device to be the temperature for inhibiting cell division;
step four: sample detection, namely sequentially detecting ten diluted samples stored in the step three, wherein the specific detection process is as follows: s1: immobilizing the amplified microspheres on a glass substrate to form a high throughput array; s2: hybridizing the universal primer with a DNA library template attached to the microsphere, and then performing a series of ligation reactions, each ligation reaction occurring between the DNA extension strand and a probe in a pool of fluorescently labeled single-stranded octanucleotide probes; s3: the base of the octanucleotide probe has a definite corresponding relation with a specific fluorescent color, after a series of complex connection, enzyme digestion and reaction cycle of the combination of the next primer, a fluorescent image is obtained, and DNA sequence information can be read out according to the corresponding relation between the base and the fluorescence, so that the number of cancer cells is obtained;
step five: counting the number of the cancer cells detected in each diluted sample in the fourth stepStatistics were performed and the number of cancer cells was recorded as C SFC And counting the specific number of cells in each sample under the electron microscope in the second step in advance, and simultaneously recording the specific number of cells in each sample as K SFC ；
Step six: calculation of cancer cells, the calculation of the number of cancer cells and all cells obtained in the fifth step was performed with reference to the following formula: p (P) SFC =C SFC /K SFC ，（P SFC Is the ratio of cancer cells);
step seven: grading cancer cells, and grading the cancer cells obtained in the step six according to the proportion P SFC Rating was performed with the following criteria: first stage: p (P) SFC If the ratio is more than or equal to 0.6 and less than 1, the cancer of cells in the sample is serious, and strong treatment is needed in a targeted manner; second stage: p (P) SFC If the ratio is more than or equal to 0.4 and less than 0.6, the canceration of cells in the sample is heavier, and strong treatment is needed selectively; third stage: p (P) SFC If the ratio is more than or equal to 0.2 and less than 0.4, the canceration intensity of cells in the sample is medium, and medium-intensity treatment is needed to be selectively carried out; fourth stage: p (P) SFC More than or equal to 0.01 and less than 0.2, the cancer strength of cells in the sample is weak, and chemotherapy and medication are selected in a targeted manner;
step eight: and (3) treatment, namely, making a treatment scheme according to the grade of the stranguria blood cancer defined in the step seven to carry out targeted treatment.
2. The method for detecting the gonococcal blood cancer developed based on high-throughput sequencing of claim 1, wherein the method comprises the following steps: the physiological saline in the step one is 0.9% sodium chloride solution, and the temperature of the physiological saline is 35.5-37.2 ℃.
3. The method for detecting the gonococcal blood cancer developed based on high-throughput sequencing according to claim 2, wherein the method comprises the following steps: the main components of the diluent in the second step are glacial acetic acid and methylene blue, and the total amount of the diluent is ten times of that of the physiological saline in the first step.
4. A method for detecting a gonomic cancer developed based on high-throughput sequencing of claim 3, wherein: in the second step, in the process of cutting the initial sample, a fine cutter is used for removing redundant tissues in the initial sample.
5. The method for detecting the gonococcal blood cancer developed based on high-throughput sequencing of claim 4, wherein the method comprises the following steps: in the sample storage process in the third step, the storage time of the diluted sample is recorded, and the corresponding deletion of the content of each cell in the diluted sample is carried out according to the division speed of cancer cells and common cells.
6. The method for detecting the gonococcal blood cancer developed based on high-throughput sequencing of claim 5, wherein the method comprises the following steps: the glass substrate in the fourth step comprises a plurality of separation areas, and no more than two diluted samples are arranged in each separation area, so that mutual pollution among the diluted samples is avoided to influence the detection result.
7. The method for detecting the gonococcal blood cancer developed based on high-throughput sequencing of claim 6, wherein the method comprises the following steps: the cancer cell proportion rating in the step seven is derived through a written report or an electronic report, and the attached treatment guidance suggestion is defined by referring to the rating content.
8. The method for detecting the gonococcal blood cancer developed based on high-throughput sequencing of claim 7, wherein the method comprises the following steps: and D, performing subsampling detection on the patients with the rated grades of the first grade and the fourth grade in the step seven so as to ensure the accuracy of data.
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