CN105567837A - 检测K-ras基因突变的引物和探针体系、方法及试剂盒 - Google Patents

检测K-ras基因突变的引物和探针体系、方法及试剂盒 Download PDF

Info

Publication number
CN105567837A
CN105567837A CN201610071654.2A CN201610071654A CN105567837A CN 105567837 A CN105567837 A CN 105567837A CN 201610071654 A CN201610071654 A CN 201610071654A CN 105567837 A CN105567837 A CN 105567837A
Authority
CN
China
Prior art keywords
primer
dna
detected
probe
ras
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.)
Granted
Application number
CN201610071654.2A
Other languages
English (en)
Other versions
CN105567837B (zh
Inventor
陈晓琦
张传雷
王新亭
张闪闪
刘姝
陈欣菊
冀爱英
刘香丽
张宇
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.)
FIRST AFFILIATED HOSPITAL OF HENAN INSTITUTE OF CHINESE MEDICINE
Original Assignee
FIRST AFFILIATED HOSPITAL OF HENAN INSTITUTE OF CHINESE MEDICINE
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 FIRST AFFILIATED HOSPITAL OF HENAN INSTITUTE OF CHINESE MEDICINE filed Critical FIRST AFFILIATED HOSPITAL OF HENAN INSTITUTE OF CHINESE MEDICINE
Priority to CN201610071654.2A priority Critical patent/CN105567837B/zh
Publication of CN105567837A publication Critical patent/CN105567837A/zh
Application granted granted Critical
Publication of CN105567837B publication Critical patent/CN105567837B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

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
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • 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
    • C12Q1/6858—Allele-specific amplification
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Landscapes

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

Abstract

本发明涉及一种检测K<b>-</b>ras基因突变的引物和探针体系，包括SEQ？ID？NO：1～13所示的核苷酸序列；还公开了其检测方法，包括配置50μL荧光PCR反应体系、进行荧光PCR扩增程序、结果判定等步骤。本发明可同时检测KRAS基因7种突变；最多可以检测1～10拷贝的基因突变；特异性强，在10～30ng？野生型DNA背景下不受干扰；检测性能突出，在10ng？DNA背景下，可以检测出1～10拷贝的基因突变。

Description

检测K-ras基因突变的引物和探针体系、方法及试剂盒
技术领域
本发明属于基因工程技术领域，具体涉及一种检测K-ras基因突变的引物和探针体系、方法及试剂盒。
背景技术
K-ras基因是EGFR信号通路中的重要分子之一，是首个影响结直肠癌临床治疗决策的生物标记物，其所编码的K-ras蛋白为EGFR信号通路下游区的一种分子G蛋白，当K-ras基因发生突变后，会导致该信号通路异常活化，从而对EGFR单抗治疗无效。因此，K-ras基因状态与针对EGFR的靶向药物（例如西妥昔单抗和帕尼单抗等）的疗效有关，《09年NCCN非小细胞肺癌临床实践指南》明确指出：如果K-ras基因发生了突变，则不建议病人使用特罗凯(Tarceva/厄洛替尼/Erlotinib)进行分子靶向治疗。
许多肿瘤中K-ras基因的突变率较高，在白血病、肺癌、直肠癌、胰腺癌等癌症中，K-ras突变很常见，尤其是在结直肠癌和非小细胞肺癌中，突变率甚至可达30～60%和21～28%。同时在上述两种肿瘤中约80～90%的K-ras基因突变发生在突变热点2号外显子12、13密码子上，其中的7个突变热点：G12C、G12R、G12S、G12V、G12D、G12A、G13V/D占到了全部突变的90%以上。
因此，检测K-ras基因上述7个突变热点，可准确判断患者对该类治疗的预测疗效，以供临床医师参考，也能尽最大的努力筛选出有效人群，避免错过治疗。
发明内容
针对上述问题，本发明提供一种检测人K-ras基因突变的引物和探针体系，可同时检测KRAS基因7种突变，最多可以检测1～10拷贝的基因突变，特异性强，在10～30ng野生型DNA背景下不受干扰，检测性能突出，在10ngDNA背景下，可以检测出1～10拷贝的基因突变。
为解决以上技术问题，本发明通过以下技术方案实现：
设计一种检测K-ras基因突变的引物和探针体系，包括SEQIDNO：1～13所示的核苷酸序列，所述引物或探针具有以下特点：
（1）本发明引进LNA修饰的野生探针封闭野生模板，探针3’-末端用PO4修饰或NH2修饰封闭阻止TaqDNA聚合酶的延伸；
（2）本发明特异性引物3’-末端碱基采用LNA修饰；
（3）本发明特异性引物3’-末端起第二个碱基引入脱氧次黄嘌呤核苷修饰。
设计一种检测K-ras基因突变的方法，包括以下步骤：
（1）合成所述引物和探针；
（2）配置50μL荧光PCR反应体系：Buffer（10×）5μL，Mg2+1.0～5mmol/L，dNTP100～1000nmol/L，上述每个引物100～500nmol/L，上述每个探针100～1000nmol/L，TaqDNA聚合酶1U～3U，DNA模板5～10ng；
（3）进行荧光PCR扩增程序：酶激活，95℃10min；突变富集，95℃30s，68℃30，64℃50s，72℃25s，16个循环；扩增检测，30个循环，95℃30s，58℃32s，72℃18s，于“58℃32s”该步骤收集荧光信号FAM和HEX；
（4）结果判定：检测体系中，HEX通道15<Ct<22，表明上样在可控范围内，结果有效；以FAM信号通道为阳性判断标准，曲线呈“S”曲线，且Ct<29为阳性，Ct为0则为阴性。
所述DNA模板包括FFPE组织及外周血游离DNA。
利用上述引物和/或探针中的至少一种，可制成用于检测K-ras基因突变的试剂盒。
本发明具有以下积极有益技术效果：
（1）本发明改良现有扩增受阻检测体系，摈弃ARMS引物设计中引入错配的原则，将特异性检测突变引物3’-末端起第2碱基采用脱氧次黄嘌呤核苷代替。脱氧次黄嘌呤核苷（dI）是天然存在的碱基，与A、G、C、T结合力弱，当与其它碱基结合时，会比其它碱基错配相对更稳定。脱氧次黄嘌呤与其它碱基的结合能力为：dI：dC>dI：dA>dI：dG>dI：dT，在DNA聚合酶的催化下，脱氧次黄嘌呤优先与dC结合。
（2）本发明在特异性检测突变引物的3’-末端碱基采用LNA修饰，提高了检测的特异性和敏感性。
（3）本发明引入了LNA修饰的封闭野生型的LNA探针，使得可以在高背景下检测低拷贝。
因此，本发明可同时检测KRAS基因7种突变；最多可以检测1～10拷贝的基因突变；特异性强，在10～30ng野生型DNA背景下不受干扰；检测性能突出，在10ngDNA背景下，可以检测出1～10拷贝的基因突变。
附图说明
图1为本发明阳性结果判定标准对照示意图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合具体实施例，对本发明进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。以下实施例中所涉及的原料，如无特别说明均为市售，所涉及检测方法如无特别说明，则均为常规方法。
实施例1
一种检测KRAS基因7种突变的试剂盒，包括SEQIDNO：1～13所示的核苷酸序列，见表1。
表1引物和探针
。
实施例2
一种检测K-ras基因7种突变的方法，包括以下步骤：
（1）配置50μL荧光PCR反应体系：Buffer（10×）5μL，Mg2+2.0mmol/L，dNTP300nmol/L，实施例1每个引物200nmol/L，实施例1每个探针300nmol/L，TaqDNA聚合酶2U，DNA模板10ng，余量为ddH2O；
（2）进行荧光PCR扩增程序：酶激活，95℃10min；突变富集，95℃30s，68℃30，64℃50s，72℃25s，16个循环；扩增检测，30个循环，95℃30s，58℃32s，72℃18s，于“58℃32s”该步骤收集荧光信号FAM和HEX；
（3）结果判定：检测体系中，HEX通道15<Ct<22，表明上样在可控范围内，结果有效；以FAM信号通道为阳性判断标准，曲线呈“S”曲线，且Ct<29为阳性，Ct为0则为阴性。
实施例3
一、K-ras突变检测体系和阈值的确定
（1）突变检测体系：通过长期的反复测试研究，本发明确定K-ras检测的上样量为10ng，将K-ras基因7种突变分别装入八排管的7个孔，第8个为外控；
（2）阳性判定：阳性判定需要满足以下要求：
首先，PCR阴性对照没有扩增曲线，有标明有污染，结果不可靠；
其次，检测103copies阳性突变，Ct<23，否则性能不稳定；
第三，外控的Ct值为：17～25，确保上样量的准确性；
最后，检测样品，曲线成“S”，且Ct<30。
满足上述条件，待检测样品K-ras基因突变为阳性。
结果无论有无突变，内参HEX通路都有曲线可以观察到，如果没有，则表明实验存在错误，需要重新实验，见图1。
二、检测结肠癌K-ras基因突变
62例结肠癌样本，已经采用厦门艾德ADX-ARMS试剂盒标定，来评价本发明实施例2检测临床样本的准确性。检测有出入或阳性的样本采用COLD-PCR后，亚克隆测序鉴定。
表2本发明检测与ADX-ARMS结果对比
通过与对62例临床样本的检测，结果与厦门艾德完全一致，且主要为Gly12Ser（GGT>AGT）、Gly12Val（GGT>GTT）、Gly12Asp（GGT>GAT）、Gly12Ala（GGT>GCT）和Gly13Asp（GGC>GAC），具体的结果参加表2。对两种方法检测判定为阳性的样本采用COLD-PCR测序，结果完成正确。在结肠癌中，K-ras的突变率约为32.8%左右。
SEQUENCELISTING
<110>河南中医学院第一附属医院
<120>检测K-ras基因突变的引物和探针体系、方法及试剂盒
<130>/
<160>13
<170>PatentInversion3.5
<210>1
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>1
gtggtagttggagctna17
<210>2
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>2
gtggtagttggagctnc17
<210>3
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>3
gtggtagttggagctnt17
<210>4
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>4
tgtggtagttggagcna17
<210>5
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>5
tgtggtagttggagcnc17
<210>6
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>6
tgtggtagttggagcnt17
<210>7
<211>17
<212>DNA
<213>人工序列
<220>
<221>modified_base
<222>(16)..(16)
<223>I
<220>
<221>misc_feature
<222>(16)..(16)
<223>nisa,c,g,ort
<400>7
gtagttggagctggtna17
<210>8
<211>18
<212>DNA
<213>人工序列
<400>8
acctctattgttggatca18
<210>9
<211>15
<212>DNA
<213>人工序列
<400>9
tggagctggtggcgt15
<210>10
<211>27
<212>DNA
<213>人工序列
<400>10
agagtgccttgacgatacagctaattc27
<210>11
<211>19
<212>DNA
<213>人工序列
<400>11
agcctgttattttgtccta19
<210>12
<211>19
<212>DNA
<213>人工序列
<400>12
atcccaattcatacggtag19
<210>13
<211>23
<212>DNA
<213>人工序列
<400>13
tggttccttccttctggcttgtc23

Claims (4)

1.一种检测K-ras基因突变的引物和探针体系，其特征在于：包括SEQIDNO：1～13所示的核苷酸序列。
2.一种检测K-ras基因突变的方法，其特征在于：包括以下步骤：
（1）合成权利要求1所述引物和探针；
（2）配置50μL荧光PCR反应体系：Buffer5μL，Mg2+1.0～5mmol/L，dNTP100～1000nmol/L，权利要求1中所述引物100～500nmol/L，权利要求1中所述探针100～1000nmol/L，TaqDNA聚合酶1U～3U，DNA模板5～10ng；
（3）进行荧光PCR扩增程序：酶激活，95℃10min；突变富集，95℃30s，68℃30，64℃50s，72℃25s，16个循环；扩增检测，95℃30s，58℃32s，72℃18s，30个循环，于“58℃32s”该步骤收集荧光信号FAM和HEX；
（4）结果判定：检测体系中，HEX通道15<Ct<22，表明上样在可控范围内，结果有效；以FAM信号通道为阳性判断标准，曲线呈“S”曲线，且Ct<29为阳性，Ct为0则为阴性。
3.根据权利要求2所述检测K-ras基因突变的方法，其特征在于：所述DNA模板包括FFPE组织DNA或外周血游离DNA。
4.一种检测K-ras基因突变的试剂盒，包括权利要求1所述引物和/或探针中的至少一种。
CN201610071654.2A 2016-02-02 2016-02-02 检测K-ras基因突变的引物和探针体系、方法及试剂盒 Active CN105567837B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610071654.2A CN105567837B (zh) 2016-02-02 2016-02-02 检测K-ras基因突变的引物和探针体系、方法及试剂盒

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610071654.2A CN105567837B (zh) 2016-02-02 2016-02-02 检测K-ras基因突变的引物和探针体系、方法及试剂盒

Publications (2)

Publication Number Publication Date
CN105567837A true CN105567837A (zh) 2016-05-11
CN105567837B CN105567837B (zh) 2019-01-29

Family

ID=55878453

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610071654.2A Active CN105567837B (zh) 2016-02-02 2016-02-02 检测K-ras基因突变的引物和探针体系、方法及试剂盒

Country Status (1)

Country Link
CN (1) CN105567837B (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106868127A (zh) * 2017-02-20 2017-06-20 人和未来生物科技（长沙）有限公司 人kras基因多点突变单管联检荧光pcr方法、试剂盒及体系
CN106978481A (zh) * 2017-02-20 2017-07-25 人和未来生物科技（长沙）有限公司 基于arms‑pcr法检测bmp3和ndrg4基因甲基化的荧光pcr方法、试剂盒及体系
CN110982886A (zh) * 2019-12-30 2020-04-10 武汉光谷联合医学检验所股份有限公司 一种人类K-ras基因突变检测试剂盒及其应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102367478A (zh) * 2011-10-25 2012-03-07 浙江大学 用于KRAS基因突变分型的ARMS-qPCR检测试剂盒及检测方法
CN104805208A (zh) * 2015-04-30 2015-07-29 山东维真生物科技有限公司 用于检测人类kras基因7种热点突变的引物探针组合物、试剂盒及检测方法
CN105177118A (zh) * 2015-07-17 2015-12-23 陈晓琦 检测人egfr基因29种突变的引物和探针体系、方法及试剂盒

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102367478A (zh) * 2011-10-25 2012-03-07 浙江大学 用于KRAS基因突变分型的ARMS-qPCR检测试剂盒及检测方法
CN104805208A (zh) * 2015-04-30 2015-07-29 山东维真生物科技有限公司 用于检测人类kras基因7种热点突变的引物探针组合物、试剂盒及检测方法
CN105177118A (zh) * 2015-07-17 2015-12-23 陈晓琦 检测人egfr基因29种突变的引物和探针体系、方法及试剂盒

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
EVGENY A ET AL.: "Increased detection rates of EGFR and KRAS mutations in NSCLC specimens with low tumour cell content by 454 deep sequencing", 《VIRCHOWS ARCH》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106868127A (zh) * 2017-02-20 2017-06-20 人和未来生物科技（长沙）有限公司 人kras基因多点突变单管联检荧光pcr方法、试剂盒及体系
CN106978481A (zh) * 2017-02-20 2017-07-25 人和未来生物科技（长沙）有限公司 基于arms‑pcr法检测bmp3和ndrg4基因甲基化的荧光pcr方法、试剂盒及体系
CN106978481B (zh) * 2017-02-20 2020-06-16 人和未来生物科技（长沙）有限公司 基于arms-pcr法检测bmp3和ndrg4基因甲基化的荧光pcr方法、试剂盒及体系
CN110982886A (zh) * 2019-12-30 2020-04-10 武汉光谷联合医学检验所股份有限公司 一种人类K-ras基因突变检测试剂盒及其应用

Also Published As

Publication number Publication date
CN105567837B (zh) 2019-01-29

Similar Documents

Publication Publication Date Title
Weinberg et al. Molecular profiling of biliary cancers reveals distinct molecular alterations and potential therapeutic targets
Zaliova et al. Genomic landscape of pediatric B-other acute lymphoblastic leukemia in a consecutive European cohort
Yang et al. IDH mutant lower grade (WHO Grades II/III) astrocytomas can be stratified for risk by CDKN2A, CDK4 and PDGFRA copy number alterations
Morgenstern et al. The challenge of defining “ultra‐high‐risk” neuroblastoma
Vanni et al. The current state of molecular testing in the BRAF-mutated melanoma landscape
Vigneswaran et al. Beyond the World Health Organization grading of infiltrating gliomas: advances in the molecular genetics of glioma classification
Hu et al. Fluorescence in situ hybridization (FISH): an increasingly demanded tool for biomarker research and personalized medicine
Park et al. Utility of targeted deep sequencing for detecting circulating tumor DNA in pancreatic cancer patients
Nagahashi et al. Genomic landscape of colorectal cancer in Japan: clinical implications of comprehensive genomic sequencing for precision medicine
Chang et al. Comprehensive molecular and clinicopathologic analysis of 200 pulmonary invasive mucinous adenocarcinomas identifies distinct characteristics of molecular subtypes
Gorovets et al. IDH mutation and neuroglial developmental features define clinically distinct subclasses of lower grade diffuse astrocytic glioma
Dietel et al. Predictive molecular pathology and its role in targeted cancer therapy: a review focussing on clinical relevance
Jansen et al. Molecular pathology in adult gliomas: diagnostic, prognostic, and predictive markers
Lan et al. Mutations in the RAS and PI3K pathways are associated with metastatic location in colorectal cancers
Bibault et al. Next-generation sequencing of FLT3 internal tandem duplications for minimal residual disease monitoring in acute myeloid leukemia
Sha et al. Association study of the let-7 miRNA-complementary site variant in the 3′ untranslated region of the KRAS gene in stage III colon cancer (NCCTG N0147 Clinical Trial)
Yamauchi et al. Serial profiling of circulating tumor DNA for optimization of anti‐VEGF chemotherapy in metastatic colorectal cancer patients
Cruz et al. Analysis of KIAA1549–BRAF fusion gene expression and IDH1/IDH2 mutations in low grade pediatric astrocytomas
Barrett et al. Clinical study of genomic drivers in pancreatic ductal adenocarcinoma
van't Erve et al. Metastatic colorectal cancer treatment response evaluation by ultra-deep sequencing of cell-free DNA and matched white blood cells
Hwang et al. Distinct mutational profile and immune microenvironment in microsatellite-unstable and POLE-mutated tumors
Kim et al. Circulating tumor DNA shows variable clonal response of breast cancer during neoadjuvant chemotherapy
CN105567837A (zh) 检测K-ras基因突变的引物和探针体系、方法及试剂盒
Tafe et al. Clinical genotyping of non–small cell lung cancers using targeted next-generation sequencing: utility of identifying rare and co-mutations in oncogenic driver genes
Huvila et al. Molecular subtype diagnosis of endometrial carcinoma: Comparison of the next-generation sequencing panel and Proactive Molecular Risk Classifier for Endometrial Cancer classifier

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant