WO2013053182A1 - 检测核酸样本中预定事件的方法和系统以及捕获芯片 - Google Patents
检测核酸样本中预定事件的方法和系统以及捕获芯片 Download PDFInfo
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- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Definitions
- the invention relates to the field of biomedicine.
- the invention relates to methods and systems for detecting predetermined events in nucleic acid samples and capture chips. Background technique
- Monogenic disorders are diseases or pathological traits controlled by a pair of alleles, also known as Mendelian diseases or monogenic genetic diseases, which can be genetically classified into autosomal recessive genetic diseases (AR), often Chromosomal dominant genetic disease (AD), X-linked recessive genetic disease (XR), X-linked dominant inheritance (XD), and Y-linked genetic disease; according to data published on the Human Genome Project Information website, there are 6,000 known species.
- a single-gene genetic disease with clinical symptoms and a clear genetic mechanism http: ⁇ www.ncbi.nlm.nih.gov/omim).
- the present invention aims to solve at least one of the technical problems existing in the prior art. To this end, it is an object of the present invention to provide a method for efficiently detecting a predetermined event in a nucleic acid sample.
- the invention proposes a method of detecting a predetermined event in a nucleic acid sample.
- the method of detecting a predetermined event in a nucleic acid sample comprises the steps of: constructing a sequencing library for the nucleic acid sample; sequencing the sequencing library to obtain a sequencing result composed of a plurality of sequencing data; Sequencing data from a predetermined region; and determining the occurrence of the predetermined event based on the composition of the sequencing data from the predetermined region.
- the above method can effectively detect a predetermined event in a nucleic acid sample, for example, can effectively detect a mutation type in a SNP site, or can effectively perform aneuploidy of a prenatal chromosome.
- the invention proposes a system for detecting a predetermined event in a nucleic acid sample.
- the system for detecting a predetermined event in a nucleic acid sample comprises: a library construction device, the library construction device being adapted to construct a sequencing library for the nucleic acid sample; a sequencing device, the sequencing device and the Library construction devices are coupled and adapted to sequence the sequencing library to obtain sequencing results consisting of multiple sequencing data; An analysis device adapted to select sequencing data from the predetermined region from the sequencing results, and to determine the occurrence of the predetermined event based on the ratio of the sequencing data from the predetermined region to the total sequencing data.
- the method for detecting a predetermined event in a nucleic acid sample described above can be effectively implemented, thereby effectively detecting a predetermined event in a nucleic acid sample, for example, can effectively detect a mutation type in a SNP site, or can be effective
- the aneuploidy of prenatal chromosomes is performed.
- the invention proposes a capture chip.
- the capture chip includes: a chip body; and a plurality of oligonucleotide probes disposed on a surface of the chip body, wherein the oligo Nucleotide probes are specific for a predetermined region in the human genome.
- the oligonucleotide probe based on the capture chip is specific to a predetermined region in the human genome, and thus, the capture chip can be effectively applied to the aforementioned method for detecting a predetermined event in a nucleic acid sample, effectively determining from a predetermined schedule.
- the sequencing data of the region enables efficient detection of predetermined regions in the human genome.
- FIG. 1 is a schematic structural diagram of a system for detecting a predetermined event in a nucleic acid sample according to an embodiment of the present invention
- FIG. 2 is a schematic structural view of a system for detecting a predetermined event in a nucleic acid sample according to still another embodiment of the present invention
- the SNP is detected, and according to the base probability distribution when the mother is heterozygous for the fetus, the simulated frequency of each base at different sequencing depths is randomly generated, and the Bayesian model shown in Formula I is used to calculate the difference.
- FIG. 5 is a schematic structural view of a capture chip according to an embodiment of the present invention. detailed description
- the invention proposes a method of detecting a predetermined event in a nucleic acid sample.
- predetermined event refers to a mutation or abnormality that may be present in a nucleic acid sample, such as a genetic variation (http://en.wikipedia.org/wiki/Genetic_variation). The site or region of occurrence of these mutations or abnormalities has been previously known or reported.
- the predetermined event that can be detected may be a structural variation of the nucleic acid sequence such as deletion, insertion, mutation, repetition, ectopic And inversion, etc., may also be a variation in the number of chromosomes such as aneuploidy, or may be a molecular genetic marker including a single nucleotide polymorphism (SNP), a small satellite, and a microsatellite sequence (STR) in the genome. .
- SNP single nucleotide polymorphism
- STR microsatellite sequence
- the inventors have discovered that the specific region of a nucleic acid sample containing a site at which a predetermined event may occur can be detected, and the sequencing results of these specific regions can be formed (for example, at a specific site, the frequency at which each ATGC base appears) Performing the analysis can effectively determine whether the predetermined event or the type of the predetermined event described above occurs in the nucleic acid sample, for example, the type of the SNP can be determined. It should be noted that, based on the method of the present invention, based on the judgment of whether a "predetermined event" occurs, further analysis may be performed on the detected results, and further conclusions may be obtained, for example, according to the implementation of the present invention.
- the method can be further applied to achieve an effective paternity test.
- predetermined event shall be understood broadly to include not only items that can be directly derived from sequencing results, but also items that are obtained by further analysis of the test results, such as determining different nucleic acids. The kinship between the samples.
- the method of detecting a predetermined event in a nucleic acid sample can include the following steps:
- the type of the nucleic acid sample is not particularly limited and may be deoxyribonucleic acid (DNA) or ribonucleic acid (RA), preferably DNA.
- DNA deoxyribonucleic acid
- RA ribonucleic acid
- the source of the nucleic acid sample is also not particularly limited.
- the nucleic acid sample that can be used is at least one selected from the group consisting of a human genomic DNA sample and a free nucleic acid.
- the genomic DNA sample is genomic DNA derived from human leukocytes or maternal plasma.
- the inventors have found that with the methods of the invention, specific events such as nucleic acid mutations in the human genome can be efficiently determined.
- specific events such as nucleic acid mutations in the human genome can be efficiently determined.
- the genetic characteristics of the fetus can be effectively analyzed to achieve prenatal diagnosis or paternity testing for the fetus.
- sequencing libraries for nucleic acid samples, those skilled in the art can appropriately select according to different sequencing technologies.
- a nucleic acid sample from a biological sample
- the method and apparatus for taking a nucleic acid sample are also not particularly limited, and can be carried out using a commercially available nucleic acid extraction kit.
- the sequencing library is applied to a sequencing instrument, the sequencing library is sequenced, and corresponding sequencing results are obtained, which are composed of a plurality of sequencing data.
- the method and apparatus that can be used for sequencing according to embodiments of the present invention are not particularly limited, and include, but are not limited to, dideoxy chain termination method; preferably high-throughput sequencing methods, whereby high utilization of these sequencing devices can be utilized
- the characteristics of flux and deep sequencing further improve the efficiency of determining the aneuploidy of nucleated red blood cells. Thereby, the subsequent analysis of the sequencing data is improved, especially the accuracy and accuracy of the statistical test analysis.
- the high throughput sequencing methods include, but are not limited to, second generation sequencing techniques or single molecule sequencing techniques.
- the second generation sequencing platform (technology) (see Metzker ML. Sequencing technologies-the next generation. Nat Rev Genet. 2010 Jan; ll(l): 31-46, which is incorporated herein by reference in its entirety) Not limited to Illumina-Solexa (GA TM , HiSeq2000TM, etc.), ABI-Solid and Roche-454 (pyrophosphate sequencing) sequencing platforms; single molecule sequencing platforms (technologies) including but not limited to Helicos's true single molecule sequencing technology (True Single Molecule DNA sequencing ) , Pacific Biosciences single molecule real-time (SMRTTM), and nanopore sequencing technology from Oxford Nanopore Technologies (see Rusk, Nicole (2009-04-01). Cheap Third-Generation Sequencing. Nature Methods 6 (4): 244-245, which is incorporated herein in its entirety by reference.
- the whole genome sequencing library can be sequenced using at least one selected from the group consisting of Illumina-Solexa, ABI-SOLiD, Roche-454, and single molecule sequencing devices.
- the obtained sequencing results are processed to determine sequencing data from a predetermined region.
- predetermined region shall be taken broadly and refers to any region of a nucleic acid molecule that contains a site at which a predetermined event may occur. For SNP analysis, it can refer to a region containing a SNP site.
- the predetermined region refers to the full length or portion of the chromosome to be analyzed, i.e., all sequencing data from that chromosome is selected.
- the method of selecting the sequencing data from the corresponding region from the sequencing results can be not particularly limited.
- sequencing data from a predetermined region can be obtained by aligning all of the obtained sequencing data with a known nucleic acid reference sequence.
- the sequencing of the sequencing library to be sequenced can also be completed before the sequencing operation, so that the sequencing data from the predetermined region can be directly obtained.
- determining sequencing data from a predetermined region may include, after obtaining the sequencing result, screening the sequencing result by a comparison method to obtain sequencing data from a predetermined region. It is also possible to select a sequencing library by sequencing before it is finally obtained from a predetermined region. The sequencing results of the sequencing data.
- the method of selecting a sequencing library is not particularly limited, and may be performed at any stage of constructing a sequencing library, for example, using a probe of a predetermined region specificity.
- a genome can be interrupted to obtain a DNA fragment, a DNA probe can be screened using a specific probe, and a subsequent library construction operation can be performed on the selected DNA fragment, thereby obtaining a sequencing library from a predetermined region.
- a subsequent library construction operation can be performed on the selected DNA fragment, thereby obtaining a sequencing library from a predetermined region.
- the method before the sequencing library is sequenced, the method further includes the step of selecting the sequencing library by using a probe, wherein the probe is specific to the predetermined region. .
- the sequencing library can be initially screened before sequencing, thereby increasing the proportion of the data that can be directly analyzed in the obtained sequencing data, and further increasing the sequencing depth, thereby simultaneously performing multiple predetermined regions of the nucleic acid sample.
- Sequencing and analysis the form of the probe is not particularly limited.
- the probe is arranged on a chip. Thus, by placing the probe on the chip, it is possible to further improve the efficiency of detection and analysis of the nucleic acid sample by realizing high-throughput screening of a plurality of predetermined regions of the sequencing library.
- probes for screening a plurality of SNP sites can be integrated on one chip, and a plurality of different diseases can be simultaneously detected by one hybridization reaction.
- the detection method of the embodiment of the present invention can detect a large number of SNP sites simultaneously, thereby realizing effective paternity testing and improving the effectiveness and timeliness of paternity testing.
- the chromosomal abnormality can be detected by the detection method of the embodiment of the present invention by using the above-described chip for detecting a single-gene disease, for example, the chromosome is effectively realized in the embodiment of the present invention. Detection of aneuploidy such as trisomy 21.
- a plurality of samples can be simultaneously detected as long as a different and known sequence of tags is added during the process of constructing a library for each sample. It greatly improves the throughput of detection, reduces the operation process and reagent loss of multiple detections in clinical applications, saves time and reduces costs, and provides great support for large-scale clinical non-invasive prenatal screening work in the future.
- a method of determining sequencing data from a predetermined region by comparison may also be combined with a method of sequencing a predetermined region by a probe, thereby improving selection of sequencing data from a predetermined region.
- the accuracy For relatively short detections of predetermined regions, for example for detection of a type of SNP mutation, it is possible to rely solely on probe hybridization screening libraries for screening of sequencing data.
- the selection of the sequencing result further includes removing the result of poor sequencing quality from the sequencing result, and in this regard, those skilled in the art can perform filtering according to predetermined criteria.
- the method further comprises: comparing the sequencing result with a known nucleic acid sequence to obtain a unique alignment sequence; And selecting sequencing data from the predetermined region from the unique alignment sequence.
- the occurrence of the predetermined event can be judged based on the composition of the sequencing data from the predetermined region.
- composition of sequencing data means that, for the region under study, all sequencing data, including the sequencing results of all the sites obtained, and the corresponding results The number of readings (reads). The inventors propose that the composition of these sequencing data can be analyzed by statistical analysis methods to eliminate accidental errors, thereby obtaining the sequencing results most likely to reflect the real situation.
- the inventors have proposed an analysis method for SNPs.
- the predetermined region is a nucleic acid fragment containing a known SNP
- the predetermined event is a mutation type of the SNP site
- determining that the predetermined event occurs in the nucleic acid sample further comprises: determining The SNP sites are the ratio of the sequencing data of bases VIII, T, G, and C, respectively, to the total sequencing data; and based on the ratio, the Bayesian model is used to determine the base with the highest probability of occurrence at the SNP site.
- the mutation type of the SNP in the predetermined region can be effectively determined, and the paternity test can be performed by detecting the mutation type of the plurality of SNP sites in the fetus and its parents. And this method can effectively detect multiple types of mutations and expand the scope of disease detection.
- ⁇ is the base error rate, which is the proportion of bases that are measured wrong during sequencing.
- Equation I is a Bayesian expansion that can be used to calculate the probability of current sequencing results when the predetermined region of the nucleic acid sample is a different genotype.
- the genotype with the highest probability is the actual genotype determined according to the analytical method of the present invention.
- ge «o3 ⁇ 4pe 0 is when the actual genotype is i, the current The probability of sequencing data can be determined by the formula
- Pr(genotype i
- sequence) represents the probability of occurrence of different genotypes in the current sequencing data.
- the type of the specific nucleic acid site of the sample can be effectively determined, for example, multiple SNPs can be simultaneously determined.
- the type of mutation can effectively detect the blood relationship between the samples, achieve effective paternity testing, and achieve effective detection of multiple diseases at the same time.
- the above analysis method using the Bayesian model can also be applied to the analysis of other nucleic acid variations. Different from the traditional single-site PCR method, this method not only involves more sites, but also the detection results are more reliable, and at the same time, multiple samples can be detected, and the flux is greatly increased, which simplifies the operation process to a large extent.
- the present invention also proposes a method of analyzing chromosome aneuploidy.
- the predetermined region is a first chromosome in the genome
- the predetermined event is aneuploidy of the first chromosome.
- determining the occurrence of the predetermined event based on the number of sequencing data from the predetermined region further includes the following steps:
- the ratio of the sequencing data from the first chromosome to the total sequencing data is determined, that is, the sequencing data from the first chromosome can be determined by comparing the sequencing data with the known genomic information, and the sequencing from the first chromosome is separately performed. The total amount of data, as well as the amount of total sequencing data, is compared to obtain the ratio of sequencing data from the first chromosome to the total sequencing data.
- first chromosome as used herein should be understood broadly, and it can refer to any chromosome of interest that is expected to be studied, the number of which is not limited to one chromosome, and even all chromosomes can be analyzed at the same time.
- the first chromosome is at least one selected from the group consisting of human chromosome 21, chromosome 18, chromosome 13 chromosome, X chromosome, and Y chromosome.
- a common human chromosomal disease can be effectively determined.
- the inventors of the present invention have surprisingly found that a method for determining chromosome aneuploidy according to an embodiment of the present invention can be very effectively applied to the detection of human chromosome 21, chromosome 18, chromosome 13, X chromosome and Y chromosome. Aneuploidy.
- the method for determining chromosome aneuploidy can be very effectively applied to prenatal testing of pregnant women, which can greatly shorten the time of detection and damage to pregnant women, and avoid the possibility of routine detection. Abortion risk.
- the source of the nucleic acid sample for studying chromosome aneuploidy is not particularly limited, and according to a specific example, the nucleic acid sample is genomic DNA extracted from maternal plasma. Therefore, the genetic diseases related to fetal chromosome aneuploidy are further detected under the premise of no damage to the fetus.
- the non-invasive sampling method used in the method avoids the risk of abortion caused by traditional amniocentesis and the like, and the auxiliary facilities such as ultrasound are omitted, and the sampling is simpler and more convenient.
- the ratio of the sequencing data of the first chromosome to the total sequencing data is significant compared with the normal nucleic acid sample. the difference.
- the aneuploidy of the chromosome can be effectively determined, thereby enabling effective detection of fetal hereditary diseases before delivery.
- predetermining parameter refers to the operation and analysis performed by repeating a nucleic acid sample of a known genome normal for a single cell of a biological sample. Relevant data about specific chromosomes. Those skilled in the art will appreciate that the same sequencing conditions and mathematical methods can be used to obtain relevant parameters for a particular chromosome, as well as relevant parameters for normal cells. Here, the relevant parameters of the normal nucleic acid sample can be used as a control parameter.
- predetermined as used herein shall be understood broadly and may be determined experimentally in advance, or may be obtained by parallel experiments when performing biological sample analysis.
- the predetermined parameter is a ratio of sequencing data from the first chromosome obtained from a normal nucleic acid sample to total sequencing data.
- the difference between the ratio of the sequencing data from the first chromosome to the total sequencing data and the predetermined parameter can be expressed by any known mathematical method, for example, by comparing the ratio with a predetermined parameter, and The obtained result is compared with a threshold value, and if it is larger than the threshold value, it is determined that the nucleic acid sample is the first chromosome 3 body for the chromosome.
- the method further includes performing a Student's test on the ratio and the parameter.
- the accuracy and accuracy of the sequencing analysis results can be further improved.
- the threshold may be set to at least 1.5, such as at least 2, more preferably at least 3, after performing the T-test.
- a system 1000 for detecting a predetermined event in a nucleic acid sample includes a library construction device 100, a sequencing device 200, and an analysis device 300, in accordance with an embodiment of the present invention.
- the above-described method of detecting a predetermined event in a nucleic acid sample according to an embodiment of the present invention can be effectively implemented. The advantages of this method have been described in detail above and will not be described again.
- library construction device 100 is adapted to construct a sequencing library for a nucleic acid sample.
- a method and a flow for constructing a sequencing library for a nucleic acid sample those skilled in the art can appropriately select according to different sequencing technologies.
- a manufacturer of a sequencing instrument such as Illumina.
- the method and apparatus for extracting a nucleic acid sample from a biological sample are also not particularly limited, and can be carried out using a commercially available nucleic acid extraction kit.
- the sequencing device 200 is coupled to the library construction device 100 and is adapted to sequence the sequencing library to obtain sequencing results consisting of multiple sequencing data.
- the method and apparatus that can be used for sequencing according to embodiments of the present invention are not particularly limited.
- second generation sequencing techniques can be employed, and third generation and fourth generation or more advanced sequencing techniques can also be employed.
- at least one selected from the group consisting of Illumina-Solexa, ABI-SOLiD, Roche-454, and a single molecule sequencing device can be used for the whole gene.
- the sequencing library was sequenced.
- the system may further include a library screening device 400, in accordance with an embodiment of the present invention.
- a probe is disposed in the library screening device 400, the probe being specific to a predetermined region to perform thinning of the sequencing library using a probe.
- the sequencing library can be subjected to preliminary dilution before sequencing, thereby increasing the proportion of the directly-analyzed data in the obtained sequencing data, and further increasing the sequencing depth to achieve multiple predetermined regions of the nucleic acid sample simultaneously.
- the probe is in the form of a chip.
- the library screening device 400 described herein can be disposed in any step of library construction, either after breaking a nucleic acid sample such as genomic DNA to obtain a DNA sheet, or in a sequencing library for obtaining genomic DNA. After that, before sequencing.
- the analysis device 300 is coupled to the sequencing device 200 and is adapted to receive sequencing results from the sequencing device 200, selecting sequencing data from the predetermined region from the sequencing results, further based on the number of sequencing data from the predetermined region And determining the occurrence of the predetermined event.
- the sequencing data from the predetermined region selected from the sequencing results has been described in detail above and will not be described herein.
- the related sequence information may be pre-stored in the analysis device 300, or the analysis device 300 may be connected to a remote database (not shown) for networking operation.
- the analysis device 300 is adapted to detect and analyze SNPs.
- the predetermined region is a nucleic acid fragment containing a known SNP
- the predetermined event is a mutation type of a SNP site
- the analyzing device 300 is adapted to: determine that the base is eight at the SNP site , the ratio of the sequencing data of T, G, and C respectively to the total sequencing data; and based on the ratio, using a Bayesian model to determine the base with the highest probability of occurrence at the SNP site, in order to determine the nucleic acid sample
- the type of mutation at the SNP site is adapted to detect that the base is eight at the SNP site , the ratio of the sequencing data of T, G, and C respectively to the total sequencing data; and based on the ratio, using a Bayesian model to determine the base with the highest probability of occurrence at the SNP site, in order to determine the nucleic acid sample
- the type of mutation at the SNP site is adapted to detect and analyze SNPs.
- the analyzing device 300 can be used to analyze the aneuploidy of the chromosome, and thus, the predetermined region is the first chromosome in the genome, and the predetermined event is aneuploidy of the first chromosome, wherein the analyzing device 300 is adapted to: determine a ratio of sequencing data from the first chromosome to total sequencing data; and determine, based on a difference between the ratio and a predetermined parameter, the nucleic acid sample for the first chromosome Whether it has aneuploidy.
- the aneuploidy of the chromosome can be effectively determined, thereby enabling effective examination of the fetal hereditary disease before delivery. Measurement.
- the first chromosome is at least one selected from the group consisting of human chromosome 21, chromosome 18, chromosome 13, X chromosome, and Y chromosome.
- the analysis device 300 further comprises a T-test device (not shown) for performing a T-test on the ratio and the parameters. Thereby, the accuracy and accuracy of the sequencing analysis results can be further improved.
- the method for detecting a predetermined event in a nucleic acid sample described above can be effectively implemented, thereby effectively detecting a predetermined event in a nucleic acid sample, for example, can effectively detect a mutation type in a SNP site, or can be effective
- the analysis of aneuploidy of prenatal chromosomes was performed.
- the term "connected” as used herein shall be understood broadly and may be either directly connected or indirectly connected as long as the above functional connections are achieved.
- the invention also proposes a capture chip for the aforementioned method for detecting a predetermined event in a nucleic acid sample.
- the chip 2000 includes a chip body 2001 and a plurality of oligonucleotide probes 2002.
- the plurality of oligonucleotide probes 2002 are disposed on the surface of the chip body 2001, wherein the oligonucleotide probes are specific for a predetermined region in the human genome.
- the capture chip it is possible to efficiently capture a nucleic acid sample corresponding to a predetermined region in the sample, whereby the efficiency of the method of detecting a predetermined event in the nucleic acid sample can be effectively improved.
- the predetermined region of interest is first determined, and then the sequence of the oligonucleotide probe is determined based on the sequence characteristics of the predetermined region.
- the type of the predetermined area is not particularly limited.
- the predetermined region is a gene region associated with a disease in a human genome.
- the gene region is located on chromosomes 18, 13 or 21 of the human genome.
- the predetermined region is a nucleic acid fragment containing a known SNP.
- the chip can be utilized to simultaneously screen a large amount of SNP related information.
- Example 1 Detection of SNP locus
- the samples taken included peripheral blood of a father and a mother during pregnancy in a family.
- the cord was taken after birth and collected by an EDTA anticoagulation tube.
- the mother's peripheral blood during pregnancy 1600g, centrifuged at 4 ° C for 10 minutes, the blood cells and plasma were separated, and the plasma was further centrifuged at 16000g for 10 minutes at 4 ° C to further remove residual white blood cells.
- TIANamp Micro DNA Kit TIANGEN
- Female's peripheral blood and fetal cord blood are directly extracted from the DNA using the kit. All DNA samples obtained, except plasma DNA samples, were interrupted to a 500 bp fragment using a CovarisTM interrupter.
- the obtained DNA fragment was constructed according to the instructions provided by HiSeq2000TM sequencer manufacturer illumia®, and the sequencing library was obtained. The specific steps are as follows: End repair:
- Klenow fragment (having 5' ⁇ 3' polymerase activity and 3' ⁇ 5 'exonuclease activity) 1 ⁇
- the ligation product was recovered using a PCR purification kit (QIAGEN). The sample was finally dissolved in 32 ⁇ l of buffer.
- the PCR reaction procedure is as follows:
- the PCR product was recovered using a PCR purification kit (QIAGEN). The sample was finally dissolved in 50 ⁇ l of buffer. The constructed library was tested to the requirements of the Agilent® Bioanalyzer 2100, and the library was quantified by Q-PCR. After passing the test, the NilithGen custom-made solid phase chip 11032 I HG 19_BGI_exon_chrM_cap_HX3 (for the chip) The details are as follows. Hybridization, the hybridized product was sequenced using an illumina® HiSeq2000TM sequencer, and the number of sequencing cycles was PE101Index (ie, bidirectional lOlbp Index sequencing). The parameter setting and operation method of the instrument are carried out according to the HiSeq2000TM sequencer operating instructions provided by the manufacturer Illumina® (this manual can be obtained from http://www.illumina.com/support/documentation.ilmn).
- the genomic sequence information Hgl9 was used as a reference sequence, and a total of 7464 probes with an average length of 150 bp were designed, which covered the region of the reference genome of 1.8M.
- the Roche NimbleGen company is integrated into the miscellaneous core piece, which is 110321 HG 19_BGI_exon_chrM_cap_HX3.
- the probe design can also be handed over to the chip company to achieve the same or similar effect as long as the probe effectively covers the area.
- the amount of data obtained by sequencing is shown in Table-1.
- the sequencing depth of white blood cell samples of parents and children is about 50x, and the depth of sequencing of peripheral blood samples of mothers during pregnancy is about 300x.
- the parameter is set to ( -V 5 -S 40 -1 40 -r 1 ). Only the reads in the comparison result that are uniquely aligned to the target area of the chip are subjected to subsequent analysis.
- whole genome sequencing and chip data have been used as standard results. Therefore, all SNP loci that fall on the target region of the chip are selected as candidate sites for analysis.
- the coverage depth and the distribution of A, T, C, and G at each SNP site were counted, and the sites with lower coverage were filtered out, and finally the base distribution of the inferred site was obtained.
- the genotypes of the fetus in the parental genome and the maternal peripheral blood were extrapolated according to the Bayesian model listed in Equation I. The specific data are shown in Table-2.
- Example 2 Chromosome aneuploidy test Select a plasma sample of pregnant women who has confirmed the fetus as T21 (Twenty-one Trisomy Syndrome), and two cases of plasma samples of pregnant women with normal fetuses. Plasma DNA was extracted and the library was performed according to the method shown in Example 1. Construction, sequencing libraries were captured using the same capture chip as in Example 1, and sequenced using an Illumina® HiSeq2000TM sequencer. For abnormal chromosome number detection, the valid data obtained by sequencing are shown in Table-3. The sequencing depth of each sample was about 50x.
- the alignment procedure was consistent with the SNP genotype inference of Example 1.
- the analysis is based on the ratio of the uniqueness of the chromosomes in each chromosome to the proportion of reads in the whole genome sequencing data.
- the ratio of the normal sample is used as a control for the division, and the obtained relative reads distribution is subjected to a T test, wherein the outliers exceeding the significant limit are the number of abnormal chromosomes.
- the results are shown in Figure 4.
- the other chromosomes were within the threshold, while chromosome 21 exceeded the threshold (3), as indicated by the arrow in Figure 4. By threshold screening, the number of chromosome 21 abnormalities can be successfully detected.
- Table -3 Sampling data production
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CN201180074169.6A CN105392893A (zh) | 2011-10-14 | 2011-12-21 | 检测核酸样本中预定事件的方法和系统以及捕获芯片 |
US14/351,468 US20140249038A1 (en) | 2011-10-14 | 2011-12-21 | Method of detecting a pre-determined event in a nucleic acid sample and system thereof |
HK16103726.7A HK1215812A1 (zh) | 2011-10-14 | 2016-03-31 | 檢測核酸樣本中預定事件的方法和系統以及捕獲芯片 |
US16/023,868 US20180371539A1 (en) | 2011-10-14 | 2018-06-29 | Method of detecting a pre-determined event in a nucleic acid sample and system thereof |
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CN201110311333.2A CN102329876B (zh) | 2011-10-14 | 2011-10-14 | 一种测定待检测样本中疾病相关核酸分子的核苷酸序列的方法 |
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US16/023,868 Continuation US20180371539A1 (en) | 2011-10-14 | 2018-06-29 | Method of detecting a pre-determined event in a nucleic acid sample and system thereof |
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PCT/CN2011/084329 WO2013053180A1 (zh) | 2011-10-14 | 2011-12-21 | 一种超级芯片及其制备方法和应用 |
PCT/CN2011/084380 WO2013053182A1 (zh) | 2011-10-14 | 2011-12-21 | 检测核酸样本中预定事件的方法和系统以及捕获芯片 |
PCT/CN2011/084395 WO2013053183A1 (zh) | 2011-10-14 | 2011-12-21 | 对核酸样本中预定区域进行基因分型的方法和系统 |
PCT/CN2012/001381 WO2013053207A1 (zh) | 2011-10-14 | 2012-10-12 | 测定待检测样本中疾病相关核酸分子的核苷酸序列的方法 |
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PCT/CN2011/084395 WO2013053183A1 (zh) | 2011-10-14 | 2011-12-21 | 对核酸样本中预定区域进行基因分型的方法和系统 |
PCT/CN2012/001381 WO2013053207A1 (zh) | 2011-10-14 | 2012-10-12 | 测定待检测样本中疾病相关核酸分子的核苷酸序列的方法 |
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US (2) | US20140249038A1 (zh) |
CN (4) | CN102329876B (zh) |
HK (2) | HK1193845A1 (zh) |
TW (1) | TW201315813A (zh) |
WO (4) | WO2013053180A1 (zh) |
Families Citing this family (45)
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KR102018934B1 (ko) * | 2012-02-27 | 2019-09-06 | 도레이 카부시키가이샤 | 핵산의 검출 방법 |
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101849236A (zh) * | 2007-07-23 | 2010-09-29 | 香港中文大学 | 利用基因组测序诊断胎儿染色体非整倍性 |
CN102127819A (zh) * | 2010-11-22 | 2011-07-20 | 深圳华大基因科技有限公司 | Mhc区域核酸文库的构建方法及用途 |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7108976B2 (en) * | 2002-06-17 | 2006-09-19 | Affymetrix, Inc. | Complexity management of genomic DNA by locus specific amplification |
US20040110153A1 (en) * | 2002-12-10 | 2004-06-10 | Affymetrix, Inc. | Compleixity management of genomic DNA by semi-specific amplification |
DE602004024034D1 (de) * | 2003-01-29 | 2009-12-24 | 454 Corp | Nukleinsäureamplifikation auf basis von kügelchenemulsion |
CN101012482A (zh) * | 2007-02-12 | 2007-08-08 | 中国农业大学 | 一种筛选基因组dna中差异位点及其侧翼序列的方法 |
EP2053132A1 (en) * | 2007-10-23 | 2009-04-29 | Roche Diagnostics GmbH | Enrichment and sequence analysis of geomic regions |
CN101921841B (zh) * | 2010-06-30 | 2014-03-12 | 深圳华大基因科技有限公司 | 基于Illumina GA测序技术的HLA基因高分辨率分型方法 |
CN101921874B (zh) * | 2010-06-30 | 2013-09-11 | 深圳华大基因科技有限公司 | 基于Solexa测序法的检测人类乳头瘤病毒的方法 |
CN102329876B (zh) * | 2011-10-14 | 2014-04-02 | 深圳华大基因科技有限公司 | 一种测定待检测样本中疾病相关核酸分子的核苷酸序列的方法 |
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- 2011-12-21 WO PCT/CN2011/084395 patent/WO2013053183A1/zh active Application Filing
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101849236A (zh) * | 2007-07-23 | 2010-09-29 | 香港中文大学 | 利用基因组测序诊断胎儿染色体非整倍性 |
CN102127819A (zh) * | 2010-11-22 | 2011-07-20 | 深圳华大基因科技有限公司 | Mhc区域核酸文库的构建方法及用途 |
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CN103890189B (zh) | 2017-07-07 |
US20140249038A1 (en) | 2014-09-04 |
CN103890189A (zh) | 2014-06-25 |
HK1193845A1 (zh) | 2014-10-03 |
US20180371539A1 (en) | 2018-12-27 |
WO2013053180A1 (zh) | 2013-04-18 |
TW201315813A (zh) | 2013-04-16 |
WO2013053183A1 (zh) | 2013-04-18 |
CN103874767B (zh) | 2016-08-17 |
CN102329876B (zh) | 2014-04-02 |
CN102329876A (zh) | 2012-01-25 |
CN105392893A (zh) | 2016-03-09 |
WO2013053207A1 (zh) | 2013-04-18 |
CN103874767A (zh) | 2014-06-18 |
HK1215812A1 (zh) | 2016-09-15 |
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