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WO2025205105A1 - Procédé d'analyse génétique - Google Patents

Procédé d'analyse génétique

Info

Publication number
WO2025205105A1
WO2025205105A1 PCT/JP2025/009991 JP2025009991W WO2025205105A1 WO 2025205105 A1 WO2025205105 A1 WO 2025205105A1 JP 2025009991 W JP2025009991 W JP 2025009991W WO 2025205105 A1 WO2025205105 A1 WO 2025205105A1
Authority
WO
WIPO (PCT)
Prior art keywords
genetic analysis
analysis method
blood cells
proteinase
minutes
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
Application number
PCT/JP2025/009991
Other languages
English (en)
Japanese (ja)
Inventor
秀敏 富田
誠二 中村
由美 上田
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mitsui Chemicals Inc
Original Assignee
Mitsui Chemicals Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Mitsui Chemicals Inc filed Critical Mitsui Chemicals Inc
Publication of WO2025205105A1 publication Critical patent/WO2025205105A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • C12Q1/6874—Methods for sequencing involving nucleic acid arrays, e.g. sequencing by hybridisation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6881—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes

Definitions

  • Patent Document 1 discloses a screening test method for determining whether a prenatal fetus has an aneuploidy, such as Down syndrome, by analyzing cell-free DNA contained in maternal whole blood.
  • Patent Document 2 discloses a method for obtaining chromosomal DNA from fetal cells.
  • Patent Document 3 discloses a method for decrosslinking fixed tissue sections and obtaining nucleic acids.
  • Patent Document 4 discloses genetic analysis of placenta-derived fetal trophoblast cells.
  • Patent Document 5 discloses a method for preparing and detecting fetal cells from a sample derived from a pregnant woman using a magnetic-antibody conjugate.
  • Patent Document 6 discloses a method for dissolving fixed biological samples, specifically protein-crosslinked nucleic acids, by digesting crosslinked proteins with a protease.
  • Patent Document 7 discloses a process for separating anucleated red blood cells from a nucleated cell-enriched fraction derived from blood, and a method for detecting fetal abnormalities using fetal mesenchymal stem cells.
  • Patent Document 8 discloses a method for separating fetal cells from a sample from a pregnant woman.
  • Patent Document 9 discloses a method for isolating and detecting fetal membrane cells from maternal blood using a fetal membrane cell marker.
  • Non-Patent Document 1 discloses a method for amplifying the whole genome from a single cell.
  • Non-Patent Document 2 discloses a method for decrosslinking multiple immobilized cells.
  • Patent Document 1 EP 2473638 B1
  • Patent Document 2 WO 2018/123220
  • Patent Document 3 WO 2011/104027
  • Patent Document 4 WO 2020/245459
  • Patent Document 5 JP 2023-156347
  • Patent Document 6 JP 2023-521579
  • Patent Document 7 US 2022/0389384
  • Patent Document 8 US 2023/0295683
  • Patent Document 9 EP 4445135 A1
  • Non-patent document 2 U. Oba, K. Kohashi, Y. Sangatsuda, Y. Oda, K. Sonoda, S. Ohga, K. Yoshimoto, Y. Arai, S. Yachida, T. Shibata, T. Ito & F. Miura: An efficient procedure for the recovery of DNA from formalin-fixed paraffin-embedded tissue sections. Biol Methods Protoc. 2022 Jul 26;7(1):bpac014. doi: 10.1093/biomethods/bpac014
  • Patent Document 1 In the case of the test using cell-free DNA disclosed in Patent Document 1, the test accuracy, particularly the positive predictive value, is low, so if the test result is positive, it will be necessary to undergo highly invasive amniocentesis or chorionic villus sampling.
  • Patent Document 2 describes a method for obtaining chromosomal DNA from fetal cells, but does not describe a specific genetic analysis method.
  • Patent Document 3 describes a method for decrosslinking fixed tissue sections and obtaining nucleic acids, but does not describe a specific genetic analysis method.
  • Patent Document 4 describes genetic analysis using placenta-derived fetal trophoblast cells, but this technology cannot be applied to the analysis of fetal nucleated red blood cells.
  • placenta-derived fetal trophoblast cells exhibit mosaicism, which differs from the actual genetic information of the fetus, posing problems for tests using placenta-derived fetal trophoblast cells.
  • nucleated red blood cells exist both of fetal and maternal origin, and antibodies are subject to nonspecific adsorption, making it difficult to isolate only fetal cells using antibodies. Therefore, it is necessary to identify fetal cells, but attempting to analyze multiple cells individually results in low throughput and high costs, making it difficult to put into practical use.
  • the genetic analysis method of the first aspect of the present disclosure involves isolating fixed maternal blood cells one by one, de-crosslinking the protein and DNA of each of the isolated blood cells using a proteinase-containing buffer, extracting the de-crosslinked DNA, performing whole-genome amplification of the extracted DNA, and amplifying target sequences from the amplified whole-genome amplification product using multiple specific markers.
  • a genetic analysis method has the same configuration as the first aspect, but in addition, the specific marker is a short tandem repeat (STR) marker.
  • STR short tandem repeat
  • a genetic analysis method has the same configuration as the first or second aspect, but the STR marker corresponds to a target sequence of a gene belonging to at least one of chromosomes 13, 18, 21, and a sex chromosome.
  • a genetic analysis method includes the configuration of any one of the first to fourth aspects, and further comprises heating at 70 to 100°C during the decrosslinking.
  • a genetic analysis method has the same configuration as any one of the first to fifth aspects, except that the proteinase is proteinase K.
  • the genetic analysis method of the eighth aspect of the present disclosure has the same configuration as the seventh aspect, except that the buffer is a Tris-HCl buffer.
  • the aspects of the present disclosure are configured to provide a genetic analysis method that suppresses changes or deterioration of cells over time, such as cell loss, fusion, destruction, or death, and reduces variation in genome amplification after decrosslinking each immobilized and isolated cell.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 5.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 6.
  • 10 is a graph showing the results of amplification of a target sequence using an STR marker in sample 7.
  • 10 is a graph showing the results of amplification of a target sequence using an STR marker in sample 8.
  • 10 is a graph showing the results of amplification of a target sequence using an STR marker in sample 9.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 10.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 11.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 12.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 13.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 14.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 15.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 16.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 17.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 18.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 19.
  • 1 is a graph showing the results of amplification of a target sequence using an STR marker in sample 20.
  • FIG. 1 is a flowchart showing an overview of the steps in an embodiment of the present disclosure. First, prior to step S1, we will explain the fixation and isolation of maternal blood cells.
  • Erythroblasts include proerythroblasts, basophilic erythroblasts, polychromatic erythroblasts, and normochromatic erythroblasts. During the process of differentiation of normochromatic erythroblasts into reticulocytes, the nucleus is lost from the blood cell. Normochromatic erythroblasts usually lose the ability to divide.
  • a fixative consisting of a paraformaldehyde solution or a glutaraldehyde solution is used for fixation.
  • the paraformaldehyde concentration is 0.1 to 6% by mass, preferably 0.1 to 5% by mass, and more preferably 1 to 4% by mass, and the fixation time is 5 to 30 minutes, preferably 10 to 20 minutes, and more preferably 10 to 15 minutes.
  • the glutaraldehyde concentration is 0.001 to 0.2% by mass, preferably 0.001 to 0.1% by mass, and more preferably 0.01 to 0.08% by mass, and the fixation time is 5 to 30 minutes, preferably 10 to 20 minutes, and more preferably 10 to 15 minutes.
  • maternal blood cells are removed using the density gradient centrifugation method described above (see WO 2012/023298 A1) or a commercially available hemolyzing reagent such as OptiLyse (Beckman Coulter) to concentrate fetal nucleated red blood cells and recover them as blood cell components.
  • the recovered blood cells are dispersed in the fixative solution described above. After the fixation time has elapsed, the recovered blood cell components are dispersed in an appropriate buffer and stored until analysis.
  • blood may be collected from the mother using a blood collection tube containing fixative in advance, and red blood cells and other particles may be removed using the density gradient centrifugation method or a hemolyzing reagent described above to concentrate fetal nucleated red blood cells and recover them as blood cell components.
  • the heat treatment temperature is 40 to 70°C, preferably 50 to 60°C, more preferably 54 to 58°C, and the heating time is 10 to 120 minutes, preferably 30 to 90 minutes, and more preferably 45 to 75 minutes.
  • the buffer preferably contains a sugar. Examples of sugars include glucose, altrose, galactose, mannose, idose, fructose, sorbose, tagatose, lactose, sucrose, kojibiose, sophorose, nigerose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, and trehalose, with trehalose being more preferred.
  • the concentration of trehalose contained in the buffer is not particularly limited, but is preferably 0.001 to 1 mol/L, more preferably 0.01 to 0.4 mol/L, and particularly preferably 0.1 to 0.3 mol/L.
  • a WGA (whole genome amplification) reaction mix is added to the cell lysate solution obtained by dissociating the crosslinks in step A-2 above, and the whole genome is amplified, followed by heating to inactivate the polymerase.
  • a commercially available whole genome amplification kit can be used for the WGA reaction mix, but it is preferable to perform whole genome amplification using the WGA reaction mix used in the MDA method (multiple displacement amplification).
  • Whole genome amplification can be performed by reaction at about 30 ° C. for about 18 hours, but may be adjusted or changed as appropriate depending on the type of whole genome amplification kit or the amount or bias of genome amplification.
  • the polymerase can be inactivated by heat treatment at about 96°C for about 5 minutes, but this may be adjusted or changed as appropriate.
  • step S5 of Figure 1 the amplification product to which the primer cocktail was added in step C-1 is subjected to multi-PCR using DNA polymerase to amplify the target sequence.
  • the target sequence is preferably an STR. Furthermore, it is desirable that this STR be contained in at least one of chromosomes 13, 18, 21, and a sex chromosome.
  • the NGS sequence obtained in the step C-5 above is decoded, and the presence or absence of a genetic disease is determined based on the presence or absence of a gene sequence causing the specific genetic disease.
  • the aneuploidy of the chromosome of interest can be identified based on whether or not a trisomy-specific STR signal (triallelic pattern) is detected from the appearance pattern of the STR markers on the chromosome of interest.
  • the specific marker is preferably a short tandem repeat (STR) marker.
  • the STR marker corresponds to a target sequence of a gene belonging to at least one of chromosomes 13, 18, 21, and a sex chromosome.
  • the blood cells are fixed in a 0.01 to 6% by mass paraformaldehyde solution or a 0.001 to 0.2% by mass glutaraldehyde solution immediately after collection from the subject. Furthermore, it is preferable that the cells are heated at 70 to 100°C during the de-crosslinking. Furthermore, it is preferable that the proteinase is proteinase K. Furthermore, it is preferable that the pH is adjusted to 7 to 9 by adding a 5 to 1,000 mM buffer during the de-crosslinking. Furthermore, it is preferable that the buffer is Tris-HCl buffer.
  • peripheral blood approximately 20 mL of peripheral blood was provided from several pregnant women aged 12 to 20 weeks under appropriate management at a hospital.
  • This blood sample was subjected to density gradient centrifugation (see WO 2012/023298) to obtain a concentrate of fetal nucleated red blood cells, after which the blood cells were fixed as described below.
  • the fixed blood cells were isolated one by one using a BD cell sorter "FACSAria TM III," and then decrosslinked as described below.
  • a primer cocktail for STR markers was added to each of the whole genome amplification products of Samples 1 to 20.
  • the amount of primer cocktail added was 22 ⁇ L per 100 ng of dsDNA.
  • a total of 61 STR markers were used: 14 sets of markers for chromosome 13, 10 sets of markers for chromosome 18, 14 sets of markers for chromosome 21, and 23 sets of markers for sex chromosomes.
  • amplifying the target sequence means that 6 or more of these 61 markers can be amplified, preferably 32 or more.
  • Multi-PCR was performed on the whole genome amplification products to which the primer cocktail had been added using Taq DNA Polymerase (Funakoshi).
  • the PCR reaction consisted of 26 cycles of thermal denaturation at 95°C for 15 seconds, annealing at 55°C for 30 seconds, and extension at 72°C for 36 seconds.
  • the amplification products obtained by multi-PCR were subjected to electrophoresis and graphed by base pair number in Figures 3-1 to 3-15 (Example) and Figures 4-1 to 4-5 (Comparative Example).
  • At least 32 of the 61 STR markers were detected in the amplification products of the example, which is more than half of the markers. More specifically, of the 15 samples, 40 or more markers were detected in 13 samples, and 50 or more markers in 10 samples. Of these 10 samples in which 50 or more markers were detected, at least 11 of the 14 markers were detected on chromosome 13, at least 6 of the 10 markers were detected on chromosome 18, at least 10 of the 14 markers on chromosome 21, and at least 17 of the 23 markers on the sex chromosomes, suggesting that whole genome amplification had occurred fairly evenly.
  • sample 3 in which 49 markers were detected, 12 of the 14 markers were detected on chromosome 13, 6 of the 10 markers on chromosome 18, 13 of the 14 markers on chromosome 21, and 18 of the 23 markers on the sex chromosomes, suggesting that whole genome amplification comparable to that achieved in samples with 50 or more markers was achieved.
  • sample 13 in which 47 markers were detected, 12 of the 14 markers were detected on chromosome 13, 6 of the 10 markers on chromosome 18, all 14 markers on chromosome 21, and 15 of the 23 markers on the sex chromosomes, suggesting that whole genome amplification comparable to that achieved in samples with 50 or more markers was achieved.
  • sample 5 where a total of 42 markers were detected, only 2 of the 10 markers were detected on chromosome 18, less than half.
  • sample 6 where a total of 38 markers were detected, only 10 of the 23 markers were detected on the sex chromosomes, less than half.
  • sample 9 where a total of 32 markers were detected, only 7 of the 14 markers were detected on chromosome 13, less than half; 5 of the 10 markers were detected on chromosome 18, less than half; and 11 of the 23 markers on the sex chromosomes, less than half.
  • the paraformaldehyde concentration used as a fixative was 7% by mass, exceeding 6% by mass, which is thought to have caused excessive fixation of the cells, presumably resulting in insufficient protein degradation and nucleic acid solubilization.
  • the amount of buffer added was 1,500 mM, exceeding 1,000 mM, presumably resulting in insufficient protein degradation.
  • protein degradation was presumably insufficient because no proteinase was added or heating was performed.
  • the amplification products of the Examples all have multiple markers detected for each chromosome, and therefore can be used for testing and diagnosis when an abnormality is present in each chromosome.
  • the amplification products of the Comparative Examples all have chromosomes for which one or no markers were detected, and therefore cannot be used for testing and diagnosis when an abnormality is present in each chromosome.
  • the resulting library pool can then be subjected to NGS using NextSeq 2000 (Illumina) to decode the NGS sequence.
  • NGS NextSeq 2000
  • cells from which Y chromosome-derived reads are obtained are male and can be determined to be cells derived from the "child.”
  • the cells can be classified into two types based on the difference in the appearance pattern of the STR marker, and the STR marker appearance pattern can be compared with that of cells known to be derived from the mother, or a minority group of cells can be determined to be cells derived from the "child.”

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
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  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un procédé d'analyse génétique qui comprend : l'isolement de cellules sanguines maternelles fixes une par une; la déréticulation de l'ADN de la protéine dans un tampon contenant de la protéinase pour chacune des cellules sanguines isolées; l'extraction de l'ADN déréticulé; la soumission de l'ADN extrait à l'amplification du génome entier; et l'amplification d'une séquence cible en utilisant une pluralité de marqueurs spécifiques à partir du produit d'amplification du génome entier amplifié.
PCT/JP2025/009991 2024-03-28 2025-03-14 Procédé d'analyse génétique Pending WO2025205105A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2024054374 2024-03-28
JP2024-054374 2024-03-28

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WO2025205105A1 true WO2025205105A1 (fr) 2025-10-02

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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2015524054A (ja) * 2012-05-24 2015-08-20 ラールスル 濾過を通して生物学的サンプルから抽出された又は単離された希少細胞の多重分析のための方法
JP2017506517A (ja) * 2014-02-28 2017-03-09 ジェン−プローブ・インコーポレーテッド ホルムアルデヒドを含有する液体系細胞診用保存剤中検体から核酸を単離する方法
WO2018123220A1 (fr) * 2016-12-27 2018-07-05 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ Procédé d'obtention d'acides nucléiques de cellules foetales
WO2020245459A1 (fr) * 2019-06-07 2020-12-10 Arcedi Biotech Aps Isolation de cellules foetales à l'aide de facs
US20220389384A1 (en) * 2016-04-07 2022-12-08 Mesotex, Inc. Process for isolating nucleated cells and nucleated cell populations and uses thereof
JP2023521579A (ja) * 2020-03-31 2023-05-25 キアゲン ゲーエムベーハー 固定された生体試料からの核酸精製
JP2023528917A (ja) * 2020-06-03 2023-07-06 テンク ジェノミクス、インコーポレイテッド 細胞に由来する標的核酸を解析する方法
US20230295683A1 (en) * 2022-01-20 2023-09-21 Luna Genetics, Inc. Isolation of fetal cells

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2015524054A (ja) * 2012-05-24 2015-08-20 ラールスル 濾過を通して生物学的サンプルから抽出された又は単離された希少細胞の多重分析のための方法
JP2017506517A (ja) * 2014-02-28 2017-03-09 ジェン−プローブ・インコーポレーテッド ホルムアルデヒドを含有する液体系細胞診用保存剤中検体から核酸を単離する方法
US20220389384A1 (en) * 2016-04-07 2022-12-08 Mesotex, Inc. Process for isolating nucleated cells and nucleated cell populations and uses thereof
WO2018123220A1 (fr) * 2016-12-27 2018-07-05 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ Procédé d'obtention d'acides nucléiques de cellules foetales
WO2020245459A1 (fr) * 2019-06-07 2020-12-10 Arcedi Biotech Aps Isolation de cellules foetales à l'aide de facs
JP2023521579A (ja) * 2020-03-31 2023-05-25 キアゲン ゲーエムベーハー 固定された生体試料からの核酸精製
JP2023528917A (ja) * 2020-06-03 2023-07-06 テンク ジェノミクス、インコーポレイテッド 細胞に由来する標的核酸を解析する方法
US20230295683A1 (en) * 2022-01-20 2023-09-21 Luna Genetics, Inc. Isolation of fetal cells

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