WO2016049877A1 - Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs - Google Patents

Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs Download PDF

Info

Publication number
WO2016049877A1
WO2016049877A1 PCT/CN2014/087987 CN2014087987W WO2016049877A1 WO 2016049877 A1 WO2016049877 A1 WO 2016049877A1 CN 2014087987 W CN2014087987 W CN 2014087987W WO 2016049877 A1 WO2016049877 A1 WO 2016049877A1
Authority
WO
WIPO (PCT)
Prior art keywords
str
cpi
patriarchal
probability
child
Prior art date
Application number
PCT/CN2014/087987
Other languages
English (en)
Chinese (zh)
Inventor
蒋浩君
芦静
陈芳
蒋慧
徐讯
刘萍
Original Assignee
深圳华大基因股份有限公司
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 深圳华大基因股份有限公司 filed Critical 深圳华大基因股份有限公司
Priority to PCT/CN2014/087987 priority Critical patent/WO2016049877A1/fr
Publication of WO2016049877A1 publication Critical patent/WO2016049877A1/fr

Links

Images

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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids

Definitions

  • Paternity testing is a technique that uses forensic science, biology, and genetics, especially using genetic fingerprinting to determine whether two individuals have parental genetic relationships [1-3].
  • the existing paternity testing techniques mainly use polymerase chain reaction (PCR), restricted fragment length polymorphisms (RFLP) and short tandem repeat polymorphisms. Techniques are used to calculate the paternity index and the probability of parentality to make judgments [4].
  • the invention provides a detection method and system based on STR typing technology in non-invasive prenatal paternity testing, the method and system comprising the following steps:
  • Y chromosome STR loci Sixteen Y chromosome STR loci were selected, namely DYS438, DYS391, DYS392, DYS393, DYS456, DYS389I, DYS390, DYS389II, DYS458, DYS19, DYS385, DYS439, DYS635, Y-GATA-H4, DYS437, DYS448,
  • a detection method based on STR typing technology in non-invasive prenatal paternity testing comprising the following steps:
  • flankal primers refer to primers containing specific public linkers and sequences adapted to the second generation sequencing sequence.
  • a total of 36 sites were selected from 16 autosomal STRs, 16 Y chromosomes, and 4 X chromosomes of 8 chromosomes in self-designed primers.
  • Amplification of the fragment of interest is carried out by high throughput PCR.
  • the high-throughput PCR chip used in the present invention (provided by wafergen, the ordering site is http://www.wafergen.com/seq-ready/custom/ a total of 5184 (36*144) micro PCR reaction wells (Fig. 1 ), each reaction well can carry out a PCR reaction of 100 nL system, and only a total of 30 ng of genomic DNA is required for detecting 54 sites per sample.
  • a round of PCR reaction is carried out by using an external primer containing a barcode and an internal primer designed by the present invention.
  • the target fragment is added, and each sample is added with a barcode and a sequencing adaptor, so that the PCR product does not have to undergo the sequencing process of the sequencing library, and the second-generation high-throughput sequencing is directly performed.
  • the high-throughput PCR amplification chip used in the present invention. Each sample is tested for 36 STR sites, and 144 samples can be PCR-reacted at a time, which is equivalent to constructing 144 second-generation sequencing libraries at one time, which greatly saves the time and cost of sequencing library construction.
  • High-throughput PCR chip support The reagents and the external primers are used for amplification and establishment of the target fragment, and the specific process is as follows:
  • the amplification product obtained in the above step was centrifuged in a plate centrifuge and collected into a 500 ul EP tube, and 50 ul of 1.5 volumes of magnetic beads was added to carry out product purification.

Landscapes

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

Abstract

La présente invention concerne un procédé et un système de détection pour tests prénataux non invasifs. Le procédé comprend les étapes suivantes : 1) sélectionner les STR et concevoir des amorces; 2) amplifier les acides nucléiques obtenus à partir du père présumé et de la mère enceinte par lesdites amorces, afin d'obtenir une séquence amplifiée; 3) séquencer ladite séquence amplifiée pour obtenir des séquences de séquençage; 4) obtenir des séquences de bases correspondantes pour chaque STR par rapport auxdites séquences ci-dessus, et effectuer le typage des STR sur la base desdites séquences de bases; 5) cribler les génotypes STR obtenus; et 6) calculer les paramètres associés du test de paternité.
PCT/CN2014/087987 2014-09-30 2014-09-30 Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs WO2016049877A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2014/087987 WO2016049877A1 (fr) 2014-09-30 2014-09-30 Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2014/087987 WO2016049877A1 (fr) 2014-09-30 2014-09-30 Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs

Publications (1)

Publication Number Publication Date
WO2016049877A1 true WO2016049877A1 (fr) 2016-04-07

Family

ID=55629304

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2014/087987 WO2016049877A1 (fr) 2014-09-30 2014-09-30 Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs

Country Status (1)

Country Link
WO (1) WO2016049877A1 (fr)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110265088A (zh) * 2019-06-14 2019-09-20 孙翊鸣 一种dna转化为图案的方法
CN110499372A (zh) * 2019-09-18 2019-11-26 山西医科大学 基于高通量测序技术的多重pcr靶向捕获分型体系及试剂盒
CN110499373A (zh) * 2019-09-18 2019-11-26 山西医科大学 鉴定复杂亲缘关系的高通量str分型系统及试剂盒
CN110734982A (zh) * 2019-09-18 2020-01-31 山西医科大学 基于高通量测序技术的连锁常染色体str分型系统及试剂盒
CN110872618A (zh) * 2018-09-04 2020-03-10 北京果壳生物科技有限公司 一种基于Illumina人全基因组SNP芯片数据判断被检样本性别的方法及用途
CN111485024A (zh) * 2019-01-29 2020-08-04 深圳华大法医科技有限公司 用于个体特征确认的引物组合、及其应用
CN112342297A (zh) * 2019-08-08 2021-02-09 深圳华大法医科技有限公司 用于同时分析多个dip和str位点的复合扩增系统、方法、试剂盒及其用途
WO2021155831A1 (fr) * 2020-02-05 2021-08-12 The Chinese University Of Hong Kong Analyses moléculaires utilisant de longs fragments acellulaires pendant la grossesse
CN115064208A (zh) * 2022-07-08 2022-09-16 北京阅微基因技术股份有限公司 一种基于最大带边权重团的二代str/snp引物设计方法和系统

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101440410A (zh) * 2008-12-29 2009-05-27 无锡中德美联生物技术有限公司 18个基因座的荧光标记复合扩增检验系统
CN103451311A (zh) * 2013-09-24 2013-12-18 无锡中德美联生物技术有限公司 一种同时分析人基因组dna 26个基因座的荧光标记复合扩增的试剂盒及其使用方法和应用
CN103820559A (zh) * 2014-03-04 2014-05-28 苏州阅微基因技术有限公司 21个短串联重复序列的复合扩增体系及试剂盒

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101440410A (zh) * 2008-12-29 2009-05-27 无锡中德美联生物技术有限公司 18个基因座的荧光标记复合扩增检验系统
CN103451311A (zh) * 2013-09-24 2013-12-18 无锡中德美联生物技术有限公司 一种同时分析人基因组dna 26个基因座的荧光标记复合扩增的试剂盒及其使用方法和应用
CN103820559A (zh) * 2014-03-04 2014-05-28 苏州阅微基因技术有限公司 21个短串联重复序列的复合扩增体系及试剂盒

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LI, LI ET AL.: "Selecting of STR Markers and Calculating of PI Value for Paternity Test", CHINESE JOURNAL OF FORENSIC SCIENCES: IDENTIFICATION SCIENCES, no. 3, 31 December 2003 (2003-12-31), pages 30 - 33 *

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110872618B (zh) * 2018-09-04 2022-04-19 北京果壳生物科技有限公司 一种基于Illumina人全基因组SNP芯片数据判断被检样本性别的方法及用途
CN110872618A (zh) * 2018-09-04 2020-03-10 北京果壳生物科技有限公司 一种基于Illumina人全基因组SNP芯片数据判断被检样本性别的方法及用途
CN111485024B (zh) * 2019-01-29 2024-04-26 深圳华大法医科技有限公司 用于个体特征确认的引物组合、及其应用
CN111485024A (zh) * 2019-01-29 2020-08-04 深圳华大法医科技有限公司 用于个体特征确认的引物组合、及其应用
CN110265088A (zh) * 2019-06-14 2019-09-20 孙翊鸣 一种dna转化为图案的方法
CN112342297A (zh) * 2019-08-08 2021-02-09 深圳华大法医科技有限公司 用于同时分析多个dip和str位点的复合扩增系统、方法、试剂盒及其用途
CN112342297B (zh) * 2019-08-08 2024-01-26 深圳华大法医科技有限公司 用于同时分析多个dip和str位点的复合扩增系统、方法、试剂盒及其用途
CN110499372B (zh) * 2019-09-18 2020-05-12 山西医科大学 基于高通量测序技术的多重pcr靶向捕获分型体系及试剂盒
CN110734982B (zh) * 2019-09-18 2020-08-07 山西医科大学 基于高通量测序技术的连锁常染色体str分型系统及试剂盒
CN110734982A (zh) * 2019-09-18 2020-01-31 山西医科大学 基于高通量测序技术的连锁常染色体str分型系统及试剂盒
CN110499373A (zh) * 2019-09-18 2019-11-26 山西医科大学 鉴定复杂亲缘关系的高通量str分型系统及试剂盒
CN110499372A (zh) * 2019-09-18 2019-11-26 山西医科大学 基于高通量测序技术的多重pcr靶向捕获分型体系及试剂盒
WO2021155831A1 (fr) * 2020-02-05 2021-08-12 The Chinese University Of Hong Kong Analyses moléculaires utilisant de longs fragments acellulaires pendant la grossesse
US11371084B2 (en) 2020-02-05 2022-06-28 The Chinese University Of Hong Kong Molecular analyses using long cell-free fragments in pregnancy
GB2605736A (en) * 2020-02-05 2022-10-12 Univ Hong Kong Chinese Molecular analyses using long cell-free fragments in pregnancy
US11591642B2 (en) 2020-02-05 2023-02-28 The Chinese University Of Hong Kong Molecular analyses using long cell-free fragments in pregnancy
AU2021216616B2 (en) * 2020-02-05 2023-04-20 The Chinese University Of Hong Kong Molecular analyses using long cell-free fragments in pregnancy
GB2605736B (en) * 2020-02-05 2023-05-24 Univ Hong Kong Chinese Molecular analyses using long cell-free fragments in pregnancy
CN115064208A (zh) * 2022-07-08 2022-09-16 北京阅微基因技术股份有限公司 一种基于最大带边权重团的二代str/snp引物设计方法和系统
CN115064208B (zh) * 2022-07-08 2023-01-03 北京阅微基因技术股份有限公司 一种基于最大带边权重团的二代str/snp引物设计方法和系统

Similar Documents

Publication Publication Date Title
US20200385810A1 (en) Methods for determining fraction of fetal nucleic acids in maternal samples
WO2016049877A1 (fr) Procédés de détection et systèmes basés sur une technologie de typage de str pour des tests prénataux non invasifs
JP6760917B2 (ja) 多型カウントを用いたゲノム画分の分析
EP2728014B1 (fr) Procédé non invasif permettant de détecter une aneuploïdie chromosomique fétale
DK2376661T3 (en) SIMULTANEOUS DETERMINATION OF aneuploidy and fetal FRACTION
US9353414B2 (en) Noninvasive diagnosis of fetal aneuploidy by sequencing
US20170363628A1 (en) Means and methods for non-invasive diagnosis of chromosomal aneuploidy
JP2020512000A (ja) 胎児の染色体異常を検出する方法
AU2018201992A1 (en) Sequencing methods and compositions for prenatal diagnoses
WO2024076469A1 (fr) Procédés non invasifs d'évaluation du rejet de greffe chez les receveuses de greffe enceintes
WO2020119626A1 (fr) Procédé de test prénatal non invasif de fœtus pour maladie génétique
JP2015517317A (ja) 双子の類型を鑑定する方法とシステム
Gunel et al. Real-Time Quantitative PCR for Detection Cell Free Fetal DNA

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14903041

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 11.08.2017)

122 Ep: pct application non-entry in european phase

Ref document number: 14903041

Country of ref document: EP

Kind code of ref document: A1