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WO2018149114A1 - Procédé et dispositif permettant de déterminer une microdélétion et une microduplication dans des chromosomes fœtaux - Google Patents

Procédé et dispositif permettant de déterminer une microdélétion et une microduplication dans des chromosomes fœtaux Download PDF

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Publication number
WO2018149114A1
WO2018149114A1 PCT/CN2017/100423 CN2017100423W WO2018149114A1 WO 2018149114 A1 WO2018149114 A1 WO 2018149114A1 CN 2017100423 W CN2017100423 W CN 2017100423W WO 2018149114 A1 WO2018149114 A1 WO 2018149114A1
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Prior art keywords
microdeletion
nucleic acid
concentration
fragment
window
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Chinese (zh)
Inventor
杜伯乐
蒋馥蔓
郭宇來
韩颖鑫
高晓峘
李胜
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Guangzhou Jingke Dx Co Ltd
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Guangzhou Jingke Dx Co Ltd
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    • G—PHYSICS
    • G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
    • G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
    • G16B20/00—ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations

Definitions

  • the present invention relates to the field of biomedicine, and in particular, to methods and apparatus for determining microdeletions and microduplications in chromosomes.
  • the existing detection methods have certain limitations.
  • the detection method 1) has low precision, and a large number of false positive results will occur. Since the results are only based on the change of the proportion of fragments in a certain region, the detection results are lacking, and the effective filtering is lacking. Method, the appearance of false positives is difficult to avoid.
  • Method 2) requires probe capture and high-depth sequencing, or needs to obtain parent-source information. High-depth capture requires design of the chip, which increases the difficulty of the experiment. High-depth sequencing increases the cost, and the uncaptured part cannot. Determination.
  • Another aspect of the invention also provides an apparatus for determining microdeletion microrepetitions in a fetal chromosome, comprising:
  • micro-deletion micro-repeat fragment concentration calculating device for obtaining a concentration fm containing a micro-deletion micro-repeat fragment
  • a fetal nucleic acid concentration obtaining device for obtaining a male fetal nucleic acid concentration fy or a female fetal nucleic acid concentration fs;
  • a first filtering device for calculating rmY or rms according to the missing copy number or repeated copy number, filtering out false positives
  • a second filtering device for taking the fractional part of the rmY dmY, or the fractional part of the rms Divide dms to determine whether dmY or dms is positive, otherwise the result is filtered out;
  • the third filtering device is configured to filter the micro-deletion micro-repeat fragments according to the determination principle, and filter to obtain a fetal chromosome micro-deletion micro-repeat fragment.
  • the method and device provided by the invention can accurately determine the microdeletion microrepetition in the chromosome, and is particularly suitable for determining the microdeletion microrepetition of the fetal chromosome in the peripheral blood of the pregnant woman.
  • the invention does not require an additional chip design, saves the cost of the chip design, and makes the experimental method simple.
  • Figure 1 is a flow diagram of a method of determining fetal microdeletion microduplication in an embodiment of the invention.
  • Figure 2 is a flow diagram of a method of obtaining a concentration fm of a fragment containing a microdeletion microrepeat in the embodiment of Figure 1.
  • FIG. 3 is a flow chart of a method of obtaining a micro-deletion micro-repeat final window in the embodiment of FIG. 1.
  • FIG. 4 is a flow chart of a method of obtaining a male fetal nucleic acid concentration fy in the embodiment of FIG. 1.
  • Figure 5 is a flow diagram of a method of obtaining a female fetal nucleic acid concentration fs in the embodiment of Figure 1.
  • Figure 6 is a flow chart of a method of obtaining a predetermined range in the method of Figure 5.
  • Figure 8 is a block diagram showing the structure of an apparatus for determining microdeletion microduplication in a fetal chromosome in another embodiment of the present invention.
  • Fig. 9 is a block diagram showing the configuration of a concentration calculating apparatus containing microdeletion microrepetition fragments in the embodiment of Fig. 8.
  • FIG. 10 is a structural block diagram of an ultimate window obtaining unit in which the micro-deletion micro-repetition in the embodiment of FIG. 8 is located.
  • Figure 11 is a block diagram showing the structure of a male fetal nucleic acid concentration fy obtaining unit in the embodiment of Figure 8.
  • Figure 12 is a block diagram showing the structure of a female fetal nucleic acid concentration fs obtaining unit in the embodiment of Figure 8.
  • Figure 13 is a block diagram showing the structure of a predetermined range determining element in the embodiment of Figure 8.
  • Figure 14 is a block diagram showing the structure of a predetermined function determining element in the embodiment of Figure 8.
  • Figure 15 is a graph showing the results of 19 sample microdeletion microrepeats in Example 2.
  • Second average depth obtaining unit 113 Microdeletion microrepeat fragment concentration obtaining unit 114
  • the micro-missing micro-repetition is the ultimate window obtaining unit 115
  • First sequencing component 1151 Alignment component 1152 Length determining component 1153
  • Primary window determination component 1154 First statistical component 1155
  • Correction component 1156 First merge component 1157
  • First filter element 1158 Second merge component 1159
  • Fetal nucleic acid concentration obtaining device 120 Male fetal nucleic acid concentration fy obtaining unit 121
  • Second sequencing component 1211 First number determining component 1212
  • Filter module 12121 Second statistical component 1213
  • Average depth acquisition component 1214 Male fetal nucleic acid concentration acquisition component 1215
  • Third sequencing component 1221 Second number determining component 1222 Frequency determining component 1223
  • Female fetal nucleic acid concentration determining element 1224 Predetermined range determining component 1225 Length determination module 12251 First frequency determination module 12252 Correlation coefficient determination module 12253 Scheduled range determination module 12254 Predetermined function determining component 1226 Second frequency determination module 12261 Fitting module 12262 Ratio calculation device 130 First filter device 140 False positive judgment unit 141 Second filter device 150 Positive judgment unit 151 And value calculation device 160 Third filter device 170
  • parent sample refers herein to a biological sample obtained from a pregnant subject, eg, a woman.
  • microdeletion microrepetition refers to deletions or duplications on the chromosome that range from 1.5 kb to 10 Mb in length.
  • GC correction refers to the correction of the GC content in the sequence.
  • the present invention provides a method for determining microduplication of fetal chromosomal microdeletions, comprising:
  • the inventors have surprisingly found that the method of the present invention enables accurate determination of microdeletion microrepetitions in chromosomes, and is particularly useful for determining fetal chromosomes in peripheral blood of pregnant women. Microdeletion microrepetition.
  • the concentration fm of the microdeletion microrepetition fragment in the step S1 is obtained by the following steps:
  • d1 the total number of sequences of the primary window containing the microdeletion microrepetitions/the total number of primary windows containing the microdeletion microrepetitions;
  • the total number of primary windows and the number of sequences without the microdeletion microrepetition can be derived from a method containing a microdeletion microrepeat end window.
  • the final window has an absolute coordinate of the start and end positions. Based on the coordinates of the secondary window, the coordinates of the secondary window are found, and then it is confirmed how many primary windows are in the secondary window, and the initial and final primary windows are removed. To exclude fluctuations in the data, and then get the final primary window, calculate the total number of sequences.
  • the final window containing microdeletion microduplication is obtained by the following steps:
  • S111 Perform nucleic acid sequencing on a biological sample containing free nucleic acid to obtain a sequencing result composed of a plurality of sequencing data;
  • each of the unique alignment sequencing sequence sets can only match one position of the reference genome
  • S117 Combine a predetermined number of adjacent primary windows into a plurality of secondary windows, and determine a number of sequences in each secondary window.
  • S121 Perform a hypothesis test on the final window obtained by the final combination, and obtain an ultimate window containing the micro-deletion micro-repeat.
  • the biological sample containing the free nucleic acid is free fetal nucleic acid in the peripheral blood of the pregnant woman.
  • the nucleic acid is DNA.
  • the sequencing result comprises a length of the free nucleic acid and a base arrangement order.
  • the "length" refers to the length of the nucleic acid, and can be expressed in units of base pairs, that is, bp.
  • the sequencing is double-end sequencing, single-end sequencing or single-molecule sequencing.
  • the length of the free nucleic acid is easily obtained, which is advantageous for the subsequent steps.
  • the predetermined length in the step S114 is 1 bp to 5 M, and the predetermined number in the step S117 is 5 to 100.
  • the predetermined length is 20-40 Kb.
  • the method of GC correction comprises using local weighted regression, linear regression or logistic regression.
  • the inter-batch adjustment is to calculate a baseline for each primary window corresponding to all samples in the sequence of the sequencing, based on the number of unique alignment sequencing sequences within each primary window based on the baseline. Weighted correction.
  • the value of T1 in the step S118 comprises calculating according to a Z-test or a T-test, the filtering is filtering out the secondary window in which the T1 value is between -3-3.
  • the value of T2 in the step S119 is calculated according to a rank sum test, a symbol test or a run test, and the non-significant difference is that the T2 value of the adjacent two windows is -3-3. between.
  • the hypothesis test in the step S121 comprises calculating according to a Z test or a T test, the test threshold being defined as 3. That is, when the statistic of the test is >3 or ⁇ -3, it is determined to be the final window containing the microdeletion microrepetition.
  • the male fetal nucleic acid concentration fy in the step S2 is obtained by the following steps:
  • the total number of primary windows and the number of sequences without the microdeletion microrepetition can be derived from a method containing a microdeletion microrepeat end window.
  • the step S212 further comprises: dividing the reference genome into a plurality of primary windows according to a predetermined length, and removing the primary window in the Y chromosome whose number of unique alignment sequences is more than 5 times the number of the average sequence.
  • the primary window is a primary window adjusted by GC modification.
  • the female fetal nucleic acid concentration fs in the step S2 is obtained by the following steps:
  • the predetermined range in the step S222 is determined by the following steps:
  • S2222 setting a plurality of candidate length ranges, and respectively determining a frequency of the unique alignment sequencing sequence that appears in each candidate length range of the plurality of control samples;
  • the predetermined range is determined based on a plurality of control samples, wherein the concentration of the nucleic acid in the control sample is known, preferably, the predetermined range is based on at least 20 control samples definite.
  • control sample is a maternal peripheral blood sample of a normal male fetus with a known ratio of free fetal nucleic acid, and the nucleic acid concentration in the control sample is determined using the Y chromosome.
  • the free fetal nucleic acid concentration in the control sample is determined using the Y chromosome, i.e., by the method of the above-described male fetal nucleic acid concentration fy of the present invention.
  • the candidate length range in the S2222 spans from 1 to 300 bp, preferably from 1 to 20 bp.
  • the plurality of candidate length ranges have a step size of 1-2 bp.
  • the candidate length ranges are 1-20, 2-21,3-22, ..., respectively, wherein the span is 20 bp and the step size is 1 bp.
  • the predetermined range in the step S222 is 179 bp to 206 bp.
  • the predetermined function in the step S223 is obtained by the following steps:
  • S2232 Fitting a frequency of the unique alignment sequencing sequence within the predetermined range among the plurality of control samples with a known nucleic acid concentration to determine the predetermined function.
  • the fit is a linear fit.
  • the step S4 further includes: if the rmY ⁇ 2 is calculated according to the missing copy number or the repeated copy number is calculated to obtain rmY ⁇ 6, it is determined to be untrustworthy, and the fake is filtered. Positive result
  • rms ⁇ 2 is calculated based on the missing copy number or the repeated copy number is calculated to obtain rms ⁇ 6, it is determined to be unreliable, and the false positive result is filtered out.
  • the step S5 further comprises: if dmY ⁇ 0.13 or dmY>0.85, dmY is positive;
  • dms ⁇ 0.15 or dms > 0.791, dms is positive.
  • the determining principle in the step S7 is: if amY is between 0.95 and 1.05, the fragment of the microdeletion microrepetition is considered to be from the mother, and the fragment of the microdeletion microrepetition is filtered. ;
  • the microdeletion microrepetitive fragment is considered to be from the mother, and the microdeletion microrepetitive fragment is filtered.
  • an aspect of the present invention also provides an apparatus 100 for determining microdeletion microrepetitions in a fetal chromosome, comprising:
  • a micro-deletion micro-repeat fragment concentration calculating device 110 for obtaining a concentration fm containing a micro-deletion micro-repeat fragment
  • a fetal nucleic acid concentration obtaining device 120 for obtaining a male fetal nucleic acid concentration fy or a female fetal nucleic acid concentration fs;
  • a first filtering device 140 configured to calculate rmY or rms according to the missing copy number or repeated copy number, and filter out false positives;
  • the second filtering device 150 is configured to take the fractional part dmY of rmY, or the fractional part dms of rms, to determine whether dmY or dms is positive, or filter out the result;
  • the third filtering device 170 is configured to filter the micro-deletion micro-repeat fragments according to the determination principle, and filter to obtain a fetal chromosome micro-deletion micro-repeat fragment.
  • the micro-deletion micro-repeat fragment concentration calculating apparatus 110 further includes:
  • An initial pole window obtaining unit 111 configured to obtain a primary window containing no microdeletion microrepetition according to a primary window containing microdeletion microrepetitions, calculate a total sequence number of primary windows containing microdeletion microduplications, and microdeletion microduplications The total number of primary windows, as well as the total number of sequences of primary windows without microdeletion microduplications and the number of primary windows that do not contain microdeletion microduplications;
  • the micro-deletion micro-repetition fragment concentration calculation device 110 further includes a micro-deletion micro-repetition in the ultimate window obtaining unit 115.
  • the micro-deletion micro-repetition in the final window obtaining unit 115 includes:
  • a first sequencing component 1151 for performing nucleic acid sequencing on a biological sample containing free nucleic acid to obtain a sequencing result composed of a plurality of sequencing data
  • a length determining component 1153 for determining a length of each unique aligned sequencing sequence in the unique aligned sequencing sequence set
  • a primary window determining component 1154 for dividing the reference genome into a plurality of primary windows according to a predetermined length, the predetermined length being 1 bp - 5 M;
  • a first statistical component 1155 configured to count the number of each unique alignment sequencing sequence falling into each primary window
  • a first merging element 1157 configured to combine a predetermined number of adjacent primary windows into a plurality of secondary windows, and determine a number of sequences in each secondary window;
  • a first filter element 1158 configured to perform statistical verification on each secondary window, calculate a T1 value, and filter the secondary window according to the T1 value;
  • a second merging component 1159 is configured to perform a statistical check on the filtered secondary window, calculate a T2 value, and merge two adjacent secondary windows having no significant difference into an ultimate window according to the T2 value;
  • the micro-deletion micro-repeat final window determining component 1161 is configured to perform a hypothesis test on the final merged final window to obtain an ultimate window containing micro-deletion micro-repeats.
  • the predetermined number of the first merging elements 1157 is between 5 and 100.
  • the predetermined length is 20-40 Kb.
  • the method of GC correction in the correction element 1156 includes the use of local weighted regression, linear regression or logistic regression.
  • the inter-batch adjustment in the correction element 1156 is to calculate a baseline for each primary window corresponding to all samples in the sequenced batch, a unique alignment within each primary window based on the baseline.
  • the number of sequencing sequences is weighted and corrected.
  • the T1 value in the first filter element 1158 comprises a calculation based on a Z-test or a T-test, which filter is filtered out of a secondary window having a T1 value between -3-3.
  • the T2 value in the second combining element 1159 is calculated according to a rank sum test, a symbol check or a run test, the insignificant difference being the T2 value of the adjacent two windows at -3 Between -3.
  • the hypothesis test in the micro-deletion micro-repeat final window determining element 1161 comprises calculating from a Z-test or a T-test, the test threshold being defined as three. That is, when the statistic of the test is >3 or ⁇ -3, it is determined to be the final window containing the microdeletion microrepetition.
  • the fetal nucleic acid concentration obtaining unit 121 further includes a male fetal nucleic acid concentration fy obtaining unit 121.
  • the male fetal nucleic acid concentration fy obtaining unit 121 includes:
  • a second sequencing component 1211 for sequencing a biological sample containing free nucleic acid to obtain a sequencing result composed of a plurality of sequencing data
  • a first number determining component 1212 configured to determine, according to the sequencing result, a number of unique aligned sequencing sequences in the Y chromosome in the sample that fall within a primary window;
  • a second statistical component 1213 for counting the sum of the number of unique aligned sequencing sequences in each primary window on the Y chromosome and the total number of primary windows;
  • the first number determining element 1212 further comprises a filtering module 12121 for grouping reference genes by a predetermined length Divided into a plurality of primary windows, the primary window in which the number of unique alignment sequences in the Y chromosome is more than 5 times larger than the average number of sequences is removed.
  • the fetal nucleic acid concentration obtaining device 120 further includes a female fetal nucleic acid concentration fs obtaining unit 122.
  • the female fetal nucleic acid concentration fs obtaining unit 122 includes:
  • a third sequencing component 1221 configured to sequence a biological sample containing free nucleic acid to obtain a sequencing result composed of a plurality of sequencing data
  • a second number determining component 1222 configured to determine, according to the sequencing result, a number of unique aligned sequencing sequences whose length falls within a predetermined range in the sample;
  • a frequency determining component 1223 for determining a frequency at which a unique alignment sequencing sequence occurs within the predetermined range based on the number of unique aligned sequencing sequences whose length falls within a predetermined range;
  • a female fetal nucleic acid concentration determining element 1224 is configured to determine a female fetal nucleic acid concentration fs in the sample according to a predetermined function based on a frequency of the unique aligned sequencing sequence occurring within the predetermined range.
  • the female fetal nucleic acid concentration fs obtaining unit 122 further includes a predetermined range determining component 1225.
  • the predetermined range determining component 1225 further includes:
  • a length determining module 12251 configured to determine a length of a unique alignment sequencing sequence included in the plurality of control samples
  • the first frequency determining module 12252 is configured to set a plurality of candidate length ranges, and respectively determine a frequency of the unique aligned sequencing sequence that occurs in each candidate length range of the plurality of control samples;
  • a correlation coefficient determination module 12253 configured to generate a frequency of uniquely aligned sequencing sequences within each candidate length range based on the plurality of control samples, and in the control sample a concentration of the nucleic acid, determining a correlation coefficient between each of the candidate length ranges and a concentration of the nucleic acid in the control sample;
  • the predetermined range determining module 12254 is configured to determine, according to the value of the correlation coefficient, at least one candidate length range or a candidate length range combination as the predetermined range.
  • the predetermined range is determined based on a plurality of control samples, wherein the nucleic acid concentration in the control sample is known, preferably, the predetermined range is determined based on at least 20 control samples of.
  • control sample is a maternal peripheral blood sample of a normal male fetus with a known ratio of free fetal nucleic acid, and the free fetal nucleic acid concentration in the control sample is determined using the Y chromosome. That is, it is determined by the method of the above-described male fetal nucleic acid concentration fy of the present invention.
  • the candidate length range spans from 1 to 300 bp, preferably from 1 to 20 bp.
  • the plurality of candidate length ranges have a step size of 1-2 bp.
  • the candidate length ranges are 1-20, 2-21,3-22, ..., respectively, wherein the span is 20 bp and the step size is 1 bp.
  • the predetermined range is from 179 bp to 206 bp.
  • the female fetal nucleic acid concentration fs obtaining unit 122 further includes a predetermined function determining component 1226.
  • the predetermined function determining component 1226 includes:
  • a second frequency determining module 12261 configured to determine, in the plurality of control samples, a frequency of occurrence of a unique alignment sequencing sequence within the predetermined range, respectively;
  • a fitting module 12262 for using the plurality of control samples in the predetermined range
  • the frequency at which a unique aligned sequencing sequence occurs is fitted to a known nucleic acid concentration to determine the predetermined function.
  • the fit is a linear fit.
  • the first filtering device 140 further includes a false positive determining unit 141, configured to calculate rmY ⁇ 6 if rmY ⁇ 2 or the repeated copy number is calculated according to the missing copy number. , then judged to be untrustworthy, filtering out false positive results;
  • rms ⁇ 2 is calculated based on the missing copy number or the repeated copy number is calculated to obtain rms ⁇ 6, it is determined to be unreliable, and the false positive result is filtered out.
  • the second filtering device 150 further includes a positive determining unit 151 for determining that dmY is positive if dmY ⁇ 0.13 or dmY>0.85; or, if dms ⁇ 0.15 or dms>0.791, Then dms is positive.
  • the determining principle in the third filtering device 170 is: if amY is between 0.95 and 1.05, the fragment of the microdeletion microrepetition is considered to be from the mother, and the microdeletion is filtered. Repeated segment
  • the microdeletion microrepetitive fragment is considered to be from the mother, and the microdeletion microrepetitive fragment is filtered.
  • the reference genome is divided into a plurality of primary windows according to a predetermined length, the predetermined length being 1 bp to 5 M, preferably 20 kp to 40 kp being a predetermined length, for example (1-20 bp, 20-40 bp, 40-80 bp, 80) -100bp, 100-120bp, ...,);
  • the male fetal nucleic acid concentration fy is calculated.
  • d2 is the average depth of the primary window without the microdeletion microrepetition
  • d2 the total number of primary windows without the microdeletion microrepetition / Number of primary windows without micro-deletion micro-repeats.
  • the predetermined range is 179 bp to 206 bp.
  • the predetermined range is obtained by the following steps:
  • this embodiment uses a male fetal control sample in which the concentration of free fetal nucleic acid is determined according to the Y chromosome, ie through the above male The method of fetal nucleic acid concentration fy is determined.
  • a combination of degrees or ranges determines a correlation coefficient between each of the candidate length ranges and the concentration of the nucleic acid; wherein the correlation coefficient is calculated by correlation, including linear regression, logistic regression, local weighting, and the like.
  • the candidate length range has a span of 1-300 bp, preferably 1-20 bp.
  • the multiple candidate length ranges have a step size of 1-2 bp.
  • the predetermined function is obtained by the following steps:
  • the missing copy number is calculated by rms ⁇ 2 or the repeated copy number is calculated to obtain rms ⁇ 6, it is determined to be untrustworthy, and the false positive result is filtered out;
  • Filtering false positives is to remove the effects of multiple copies and make the results more accurate.
  • microdeletion microrepetitive fragment is considered to be from a mother, and the microdeletion microrepetitive fragment is filtered;
  • the microdeletion microrepetitive fragment is considered to be from the mother, the microdeletion microrepetitive fragment is filtered, and the fetal chromosomal microdeletion microrepetitive fragment is obtained after filtration.
  • One batch of 100 samples was selected, and 2 ml of peripheral blood was extracted for plasma separation.
  • Library construction can be performed with reference to plasma library construction requirements well known to those skilled in the art.
  • the sequencing process can be performed on a computer basis with reference to sequencing procedures well known to those skilled in the art.
  • the sequencing results are aligned to the reference genome to determine the location of the unique alignment sequence.
  • the reference genome was divided into multiple primary windows according to the length of 20 kb.
  • the number of unique alignment sequences and GC content in each primary window was counted, and the number of sequences falling into the primary window by local weighted regression was used for GC correction.
  • Table 1 Table shows the results of 19 samples, the first column is the id of the sample, the second column is the microdeletion microrepetition, and the third column is the microdeletion microrepetition length of the chromosome, fourth The column is the detected T value.
  • the selected region was 179bp-206bp, and the correlation coefficient was R -0.9056996.
  • the relationship between the frequency of the nucleic acid in the remaining 11 samples ranging from 179 bp to 206 bp in the range of 179 bp to 206 bp was calculated as a function of the free fetal nucleic acid concentration.
  • Filtration is performed according to the copy number, and the missing fragment having the copy number rmY value of 2 or more is filtered, and the fragment having the repeated copy number rmY value of 6 or more is filtered.
  • fragments having a dmY greater than 0.13 and less than 0.85 were filtered.
  • Fragments with amY > 0.95 and amY ⁇ 1.05 were filtered to obtain samples of male fetuses containing microdeletions and microduplications.
  • fragments with dms greater than 0.15 and less than 0.791 were filtered.
  • Fragments with ams > 0.93 and ams ⁇ 1.06 were filtered to obtain a sample of female fetuses containing microdeletions and microduplications.
  • the results of the micro-deletion micro-replication can be filtered to obtain a large number of false positives, and an accurate result is obtained, see FIG.
  • the abscissa indicates the number of the sample
  • the ordinate indicates the concentration
  • fm indicates the concentration estimated by the microdeletion microrepetition
  • fy indicates the concentration estimated by the male fetus sample according to chrY
  • fs indicates the concentration estimated by the fetus according to the fragment.
  • the method for determining the microdeletion microrepetition in the fetal chromosome in this embodiment is the same as that in the second embodiment. Again, the difference is that in step 4.2, the 40 kb window is divided.
  • the method for determining the microdeletion microrepetition in the fetal chromosome in this embodiment is the same as that in the second embodiment, except that the step 4.4 is performed in units of 200, and the length of the secondary window obtained after the combination is 4M.

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Abstract

L'invention concerne un procédé et un dispositif permettant de déterminer une microdélétion et une microduplication dans les chromosomes fœtaux. Le procédé comprend les étapes consistant : à acquérir la concentration fm de fragments contenant la microdélétion et la microduplication ; à acquérir la concentration d'acide nucléique fœtal mâle/femelle fy/fs ; à calculer le rapport rmY de la concentration fm de fragments contenant la microdélétion et la microduplication à la concentration d'acide nucléique fœtal mâle fy, ou le rapport rms de fm à la concentration d'acide nucléique fœtal femelle fs ; à calculer rmY ou rms selon le nombre de copies supprimées ou le nombre de copies dupliquées, et à filtrer les faux positifs ; à prendre la partie décimale dmY de rmY ou la partie décimale dms de rms, à déterminer si dmY/dms est positif ou non, et si tel n'est pas le cas, alors à filtrer le résultat ; à calculer la somme amY de la concentration fm de fragments contenant la microdélétion et la microduplication et de la concentration d'acide nucléique fœtal mâle fy, ou la somme ams de fm et de la concentration d'acide nucléique fœtal femelle fs ; et à filtrer les fragments de microdélétion et de microduplication selon un principe d'évaluation, et à obtenir les fragments de microdélétion et de microduplication des chromosomes fœtaux après filtrage.
PCT/CN2017/100423 2017-02-17 2017-09-04 Procédé et dispositif permettant de déterminer une microdélétion et une microduplication dans des chromosomes fœtaux Ceased WO2018149114A1 (fr)

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CN106778069B (zh) * 2017-02-17 2020-02-14 广州精科医学检验所有限公司 确定胎儿染色体中微缺失微重复的方法及设备
CN110970089B (zh) * 2019-11-29 2023-05-23 北京优迅医疗器械有限公司 胎儿浓度计算的预处理方法、预处理装置及其应用
CN112037846A (zh) * 2020-07-14 2020-12-04 广州市达瑞生物技术股份有限公司 一种cffDNA非整倍体检测方法、系统、储存介质以及检测设备
CN116246704B (zh) * 2023-05-10 2023-08-15 广州精科生物技术有限公司 用于胎儿无创产前检测的系统

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