Non-invasive diagnosis method for infectious diseases based on deep learning
A deep learning and disease technology, applied in neural learning methods, medical automated diagnosis, informatics, etc., can solve problems such as insufficient extraction of representative antibody library, low prediction accuracy of disease diagnosis model, etc., to improve the accuracy of model diagnosis, The effect of improving prediction accuracy
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Embodiment 1
[0036] Example 1: Training set and test set selection
[0037] Collect a certain number of pathogen-infected samples and a certain number of healthy control samples, and randomly assign all collected pathogen-infected samples and healthy control samples according to a certain ratio as a training set and a test set. The number of samples of each category is not less than 100, preferably not less than 200. The ratio is 2:1˜5:1, preferably 2.5:1˜4.5:1.
Embodiment 2
[0038] Example 2: Antibody library data acquisition
[0039] (1) Sample collection
[0040] Take 1 ml of human peripheral blood samples (Peripheral Blood Mononuclear Cells, PBMCs) from the pathogen-infected patients and healthy controls in Example 1, collect them in anticoagulant tubes containing EDTA, and store them at room temperature for no more than 4 hours. PBMCs were separated by density gradient centrifugation using lymphatic separation medium (Axis-Shield, 1114547), and the separated cells were lysed in RLT buffer (Qiagen), added with 1% β-mercaptoethanol (Sigma), and then stored at -80°C For short term storage.
[0041] (2) RNA extraction, reverse transcription
[0042] RNA was extracted using RNeasy Mini Kit (Qiagen, 74106) according to the instructions, RNA concentration was measured using NanoDrop 2000c, and 500ng of each sample was taken for reverse transcription.
[0043] Reverse transcription using Thermo SuperScript TM II Reverse Transcriptase and Takara's...
Embodiment 3
[0060] Example 3: Sequencing data processing
[0061] The downstream analysis was performed on the initial sequencing data (FASTQ file) obtained in Example 2 using MiXCR (version 3.0.7) software. Analysis with the same V / J gene and CDR3 nucleic acid sequence was defined as the same antibody molecule (clone), and only IgG antibody molecules were retained. The analysis parameters are as follows:
[0062] Align: mixcr align --library my_library -t 8 -r align_log.txt R1 R2 alignments.vdjca -s hs
[0063] Assemble: mixcr assemble -r assemble_log.txt -OseparateByV=true - OseparateByJ=true -OseparateByC=true alignments.vdjcaclones.clna
[0064] Export clones: mixcrexportClones–c IGH clones.clna clones.txt
[0065] Export Alignments: mixcrexportAlignments -f -readIds -cloneId -vHit - vAlignment -jHit -jAlignment -cHit -cAlignment -nFeature FR1 -nFeature CDR1 - nFeature FR2 -nFeature CDR2 -nFeature FR3 -nFeature CDR3 -nFeature FR4 - aaFeature FR1 -aaFeature CDR1 -aaF...
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