CN114350804A - 一种aml相关的融合基因及其应用，检测引物、试剂盒 - Google Patents
一种aml相关的融合基因及其应用，检测引物、试剂盒 Download PDFInfo
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Abstract
本发明涉及AML临床诊断技术领域，公开了一种AML相关的融合基因及其应用，检测引物、试剂盒。本发明针对高通量测序发现的白血病新融合基因SART3‑RARG，设计了特异性的PCR引物，通过实验证实该引物可用于常规RT‑PCR特异性检测，亦可用于实时荧光定量qPCR用于诊断以及定量监测，本发明扩大了原有检测手段的检测范围,能够应用于临床,可用于SART3‑RARG急性髓系白血病的诊断及疗效监测,为白血病的诊断分型及分子靶向干预提供依据。
Description
技术领域
本发明涉及AML临床诊断技术领域，具体的说是涉及一种AML相关的融合基因及其应用，检测引物、试剂盒。
背景技术
急性髓性白血病(AML)是一类来源于髓系造血干祖细胞的恶性克隆性血液系统疾病，急性早幼粒细胞白血病(APL)是AML的一个特殊类型，约占AML的10％，FAB协作组根据其细胞形态学特点将其定义为AML-M3型，其分子学特点为涉及17号染色体q21上的维甲酸受体α(RARA)与位于15号染色体q22上的早幼粒白血病基因(PML)的断裂重组，在超过95％的患者中可检测出PML-RARA融合基因，2016年世界卫生组织(WHO)因此将其定义为APL withPML-RARA。该型疾病易合并弥漫性血管内凝血及重要脏器出血，早期死亡率高。
全反式维甲酸(ATRA)通过靶向PML-RARA融合蛋白，诱导早幼粒白血病细胞分化、改善凝血功能障碍，显著降低了此类患者的早期死亡，维甲酸联合化疗或三氧化二砷已成为临床上该类患者的一线治疗，经过规范治疗80％以上患者可获长期无病生存。
维甲酸受体(RAR)属于核受体因子超家族成员，包括alpha(RARA)、beta(RARB)、gamma(RARG)三个亚型，三个亚型具有高度的序列相似性，功能上有重复亦有区别。近期陆续报道了涉及RARB或RARG的重排极少数病例，包括1种累及RARB的融合基因(TBLR1-RARB)和5种累及RARG的融合基因(NUP98-R ARG,PML-RARG,CPSF6-RARG，NPM1-RARG，HNRNPC-RARG)。这些患者都表现出AML-M3的形态学和免疫表型特点，以及出凝血障碍，但普遍对维甲酸诱导分化治疗无效，凝血功能得不到纠正，早期死亡率高，部分患者对AML化疗有效，部分学者认为这类疾病应归属为AML，因此，早期识别并调整治疗方案非常重要。
融合基因在恶性血液肿瘤的诊断、分型、微小残留病(MRD)监测及其预后，均有重要价值。累及维甲酸受体的不同亚型，虽然具有相同的细胞形态学、免疫表型特点，但同时具有一些独特的致病机理，对维甲酸治疗反应不一，因此，对RAR相关融合基因的精确诊断将有助于及早采取正确有效的干预措施，实施个体化治疗。
发明内容
本发明首要目的是提供一种AML相关的融合基因，扩展了APL样AML中RAR G相关融合基因的类型。
所述的AML相关的融合基因由SART3外显子18及其上游序列(上游序列是指SART3基因的外显子1-外显子17的序列)和RARG外显子3及其下游序列(下游序列是RARG基因外显子4-外显子10的序列)，通过SART3外显子18和RAR G外显子3直接融合形成，其序列见SEQID NO.1所示。
SART3基因序列在GeneBank中的登录号为NM_014706.4，RARG基因序列在GeneBank中的登录号为NM_000966.6；所述SART3外显子18核苷酸序列如SEQ ID NO.4所示，RARG外显子3核苷酸序列如SEQ ID NO.5所示，通过两者直接融合为SART3 exon18-RARG exon3。融合基因示意图见图1。
本发明的第二个目的是提供检测所述的AML相关的融合基因的试剂在制备A ML辅助诊断制剂中的应用。
所述的应用，辅助鉴别经典APL。进一步地，检测所述的AML相关的融合基因的试剂包括PCR检测试剂。
更进一步地，所述的PCR检测试剂中上游引物以SART3外显子18为靶基因设计，下游引物以RARG外显子3为靶基因设计。
所述引物序列优选：如SEQ ID NO.2和SEQ ID NO.3所示。
本发明的第三个目的是用于扩增所述的AML相关的融合基因的引物，所述引物序列如SEQ ID NO.2和SEQ ID NO.3所示。该引物对具有特异性和敏感性，可用于诊断及MRD监测(检测残留病变measurable residue disease)。
本发明的第四个目的是提供一种AML诊断试剂盒，包括检测所述融合基因的试剂。
进一步地，辅助鉴别经典APL。
所述的AML相关的融合基因的试剂包括PCR检测试剂。
进一步地，所述的PCR检测试剂中上游引物以SART3外显子18为靶基因设计，下游引物以RARG外显子3为靶基因设计。
更进一步地，所述引物序列如SEQ ID NO.2和SEQ ID NO.3所示。
本发明通过高通量测序技术首次发现，在一例临床表现、形态学和免疫表型符合APL的初诊患者，发现其细胞遗传学未见典型t(15；17)(q24；q21)染色体易位、未见RARA断裂、分子学上未见PML-RARA融合基因，检测出SART3-RARG融合基因，该融合基因由SART3外显子18和RARG外显子3融合形成。因此，检测出SART3-RARG融合基因表达，可以预估该患者为AML患者，并与经典APL患者相鉴别。
本发明提供的新融合基因，引物遵循设计原则，引物最好在模板cDNA的保守区内设计，长度在15bp-30bp之间，引物GC含量在40％-65％之间，退火温度接近Tm值，引物自身及引物之间不应存在互补序列。扩增实验结果显示，本发明提供的扩增引物，扩增出单一特异的电泳条带；进一步利用本发明提供的扩增引物进行检测，该引物组合诊断的特异性达到100％，敏感性达到了10-3，检测时间仅需三个工作日。
总之，本发明提供了RARG相关AML的新融合基因SART3-RARG，并针对该融合基因设计了特异性的PCR引物，扩大了原有检测手段的检测范围，能够应用于临床，可提高诊断APL样AML的检出率和准确率，为诊断分型及分子靶向治疗提供依据。
附图说明
图1为本发明融合基因的示意图；其中，箭头左边表示SART3基因外显子18及其上游序列，图中仅显示出SART3基因外显子1-18(ex1-18)；箭头右边表示RARG基因外显子3及其下游序列，图中仅显示出RARG基因外显子3-10(ex3-10)；
图2为实施例2的PCR扩增结果；
图3为实施例2扩增产物测序结果；
图4为实施例3对照组病例SART3-RARG融合基因的扩增结果；
图5为实施例3不同稀释度的SART3-RARG融合基因的扩增结果。
具体实施方式
本发明实施例公开了一种RARG相关APL样AML的融合基因及其检测引物和应用，本领域技术人员可以借鉴本文内容，适当进行工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明内。本发明所述融合基因及其检测引物和应用已通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述融合基因及其检测引物和应用进行改动或适当变更与组合，来实现和应用本发明技术。
本发明扩增引物组合是对原有APL样AML中伴随RARG融合基因检测的PCR引物的补充，扩展了APL样AML中RARG相关融合基因的类型，增加了RT-PCR方法诊断APL样AML的检出率。
以下就本发明所提供的一种RARG相关AML的融合基因及其检测引物和应用做进一步说明。
实施例1：针对明确诊断的病例进行验证和分析
患者，男性，40岁，因“牙龈出血、皮肤瘀斑20余天”就诊。血常规：白细胞计数3.18×109/L↓，血红蛋白64g/l↓，红细胞计数1.95×10^12/L↓，血小板计数48×109/L↓。凝血功能：凝血酶原时间21.0sec↑(ref 10s-14s)，血浆活化部分凝血活酶时间45.4sec↑(ref28s-45s)，血浆纤维蛋白原0.60g/l↓(ref 2.0g-4.0g)，纤维蛋白降解产物74.66ug/mL↑，D二聚体定量>40ug/mL FEU↑(ref 0-0.55ug/ml)。骨髓形态学：骨髓增生活跃，粒系占88％，红系占2％；粒系异常早幼粒细胞占73％，胞浆颗粒增多、覆盖于核上，可见内外浆；红系比例低，成熟红细胞无明显形态异常；淋巴细胞占10％，单核细胞无明显增减；巨核少，血小板少见；未见寄生虫及其他异常细胞。MPO染色阴性。免疫分型：P2占有核细胞68.3％，表达CD13,CD33,CD15,部分表达CD117、cMPO,不表达CD2,CD7,CD10,CD19,CD22,CD20,CD14,CD64,CD11b,CD38,CD9,CD56,CD34,HLA-DR,为异常早幼粒细胞；P3占有核细胞6.4％，为成熟淋巴细胞。因此，其临床表现、形态学和免疫分型基本符合APL的诊断，初步诊断为急性早幼粒细胞白血病(APL)。
但该患者的染色体结果为46，-Y,X[20]，细胞遗传学未见t(15；17)(q22；q21)染色体异常,分子生物学未检测到PML-RARA融合基因。本发明通过高通量测序技术，检测出SART3-RARG融合基因，发现该基因由SART3外显子18和RARG外显子3融合形成。
实施例2：扩增实验
对于实施例1中高通量测序RNA-seq检测出SART3 exon18-RARG exon3新融合基因的病例，使用本发明设计的引物进行验证，
表1
| SEQ ID | 序列(5’——3’) |
| NO.2 | TCAAAGTGGCAATCAGCAACC |
| NO.3 | AGCCTGGGAGGCTCCGTACC |
RNA提取：取患者新鲜骨髓液2mL,置于EDTA-K2抗凝管中，用细胞裂解液分离总白细胞，颠倒混匀，室温放置10分钟，1500rpm离心5分钟。采用TRIz ol试剂提取标本总RNA,RNA沉淀用20ul DEPC处理水溶解。经紫外分光光度计检测RNA浓度和纯度后,直接进行逆转录或置于-80℃冻存。
逆转录PCR(RT-PCR)按两步法操作步骤将RNA逆转录为cDNA。PCR扩增引物为表1引物组合。扩增体系包括5u/ulTaq酶0.2ul,10×Taq Buffer(含MgCl2)2.5uL,10mmol/L dNTP0.5ul(以上试剂均购自美国Thermo生物公司)，10umol/L的上、下游引物各0.5ul,cDNA模板150ng,无菌去离子水加至25uL。PCR扩增条件为94℃变性10min后，94℃30s、60℃30s、72℃40s,共35次循环，最后72℃延伸7min。PCR产物用2％琼脂糖，电压100V,电泳,GelRed(购自Biotium公司)染色后于紫外光下观察结果，并送测序。
扩增结果见图2,使用本发明引物PCR后可见单一特异的电泳条带，对扩增产物进行测序，得到SART3 exon18-RARG exon3融合基因融合处的序列(图3)，扩增产物的全部序列如下：
tcaaagtggcaatcagcaaccctcctcagaggaaagttccagagaagccagagaccaggaaggcaccaggtggccccatgcttttgccgcagacatacggagcgacttttggaggcccagtgggcaggccaggcagggcgggtacggagcctcccaggct，见SEQ ID NO.6。
实施例3：针对对照组病例进行检测
选取了30例诊断明确AML病例(包括10例典型APL、1例CPSF6-RARG阳性APL样AML和19例典型AML(非APL))，将这30例分别编号为C1～C30,选取3例正常人样本作为对照组病例，编号N1～N3，使用本发明的引物组合进行检测,扩增β-actin作为内参对照,RNA提取、逆转录PCR和凝胶电泳方法同实施例2,结果见表2和图4。
表2
| 标本编号 | 检测结果 | 标本编号 | 检测结果 | 标本编号 | 检测结果 | 标本编号 | 检测结果 |
| C1 | 阴性 | C11 | 阴性 | C21 | 阴性 | N1 | 阴性 |
| C2 | 阴性 | C12 | 阴性 | C22 | 阴性 | N2 | 阴性 |
| C3 | 阴性 | C13 | 阴性 | C23 | 阴性 | N3 | 阴性 |
| C4 | 阴性 | C14 | 阴性 | C24 | 阴性 | ||
| C5 | 阴性 | C15 | 阴性 | C25 | 阴性 | ||
| C6 | 阴性 | C16 | 阴性 | C26 | 阴性 | ||
| C7 | 阴性 | C17 | 阴性 | C27 | 阴性 | ||
| C8 | 阴性 | C18 | 阴性 | C28 | 阴性 | ||
| C9 | 阴性 | C19 | 阴性 | C29 | 阴性 | ||
| C10 | 阴性 | C20 | 阴性 | C30 | 阴性 |
同时，取实施例1病例(P1)标本与编号C1的对照组标本混合,进行倍比稀释，分别取1：10、1：100、1：1000、1：10000,共4个不同浓度的稀释样品，使用本发明表1的引物组合进行检测,RNA提取、逆转录PCR和测序方法同实施例2。
结果:使用本发明设计引物组合检测，30例对照组病例均未检测出SART3-R ARG融合基因，证明本发明设计的引物特异性达100％。实施例1病例标本经稀释后，在1:1000仍能检测到明显的阳性条带,1:10000未检出，证明本发明引物组合对SART3-RARG融合基因的检测敏感度达10-3(图5)。
以上所述只是用于理解本发明的方法及其核心思想，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，可以对本发明进行若干改进和修饰,这些改进和修饰也落入本发明权利的保护范围。
序列表
<110> 中南大学湘雅二医院
<120> 一种AML相关的融合基因及其应用，检测引物、试剂盒
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 4221
<212> DNA
<213> 智人(Homo sapiens)
<400> 1
atggcgactg cggccgaaac ctcggcttca gaacccgagg ctgagtccaa ggctgggccc 60
aaggctgacg gagaggagga tgaggttaag gcggctagga caaggaggaa ggtgttatcg 120
cgggctgtgg ccgctgcgac atacaagacc atggggccag cgtgggatca gcaggaggaa 180
ggcgtgagcg agagcgatgg ggatgagtac gccatggctt cctccgcgga gagctccccc 240
ggggagtacg agtgggaata tgacgaagag gaggagaaaa accagctgga gattgagaga 300
ctggaggagc agttgtctat caacgtctat gactacaact gccatgtgga cttgatcaga 360
ctgctcaggc tggaagggga gcttaccaag gtgaggatgg cccgccagaa gatgagtgaa 420
atctttccct tgactgaaga gctctggctg gagtggctgc atgacgagat cagcatggcc 480
caggatggcc tggacagaga gcacgtgtat gacctctttg agaaagccgt gaaggattac 540
atttgtccta acatttggct agagtatggc cagtactcag ttggtgggat tggtcagaaa 600
ggtggccttg agaaagttcg ctccgtgttt gaaagggctc tctcgtctgt tggtttacat 660
atgaccaaag gactcgccct ctgggaggct taccgagagt ttgaaagtgc gattgtggaa 720
gctgctcggc ttgagaaagt ccacagtctt ttccggcgac agttggcgat cccactctat 780
gatatggagg ccacatttgc agagtatgaa gaatggtcag aagacccaat accagagtca 840
gtaattcaga actataacaa agcactacag cagctggaga aatataaacc ctatgaagaa 900
gcactgttgc aggcagaggc accaaggctg gcagaatatc aagcatatat cgattttgag 960
atgaaaattg gcgatcctgc tcgcattcag ttgatctttg agcgcgccct ggtcgagaac 1020
tgccttgtcc cagacttatg gatccgttac agtcagtacc tagatcgaca actgaaagta 1080
aaggatttgg ttttatctgt acataaccgc gctattagaa actgcccctg gacagttgcc 1140
ttatggagtc ggtacctctt ggccatggag agacatggag ttgatcatca agtaatttct 1200
gtaaccttcg agaaagcttt gaatgccggc ttcatccagg ccactgatta tgtggagatt 1260
tggcaggcat accttgatta cctgaggaga agggttgatt tcaaacaaga ctccagtaaa 1320
gagctggagg agttgagggc cgcctttact cgtgccttgg agtatctgaa gcaggaggtg 1380
gaagagcgtt tcaatgagag tggtgatcca agctgcgtga ttatgcagaa ctgggctagg 1440
attgaggctc gactgtgcaa taacatgcag aaagctcggg aactctggga tagcatcatg 1500
accagaggaa atgccaagta cgccaacatg tggctagagt attacaacct ggaaagagct 1560
catggtgaca cccagcactg ccggaaggct ctgcaccggg ccgtccagtg caccagtgac 1620
tacccagagc acgtctgcga agtgttactc accatggaga ggacagaagg ttctttagaa 1680
gattgggata tagctgttca gaaaactgaa acccgattag ctcgtgtcaa tgagcagaga 1740
atgaaggctg cagagaagga agcagccctt gtgcagcaag aagaagaaaa ggctgaacaa 1800
cggaaaagag ctcgggctga gaagaaagcg ttaaaaaaga agaaaaagat cagaggccca 1860
gagaagcgcg gagcagatga ggatgatgag aaagagtggg gcgatgatga agaagagcag 1920
ccttccaaac gcagaagggt cgagaacagc atccctgcag ctggagaaac acaaaatgta 1980
gaagtagcag cagggcccgc tgggaaatgt gctgccgtag atgtggagcc cccttcgaag 2040
cagaaggaga aggcagcctc cctgaagagg gacatgccca aggtgctgca cgacagcagc 2100
aaggacagca tcaccgtctt tgtcagcaac ctgccctaca gcatgcagga gccggacacg 2160
aagctcaggc cactcttcga ggcctgtggg gaggtggtcc agatccgacc catcttcagc 2220
aaccgtgggg atttccgagg ttactgctac gtggagttta aagaagagaa atcagccctt 2280
caggcactgg agatggaccg gaaaagtgta gaagggaggc caatgtttgt ttccccctgt 2340
gtggataaga gcaaaaaccc cgattttaag gtgttcaggt acagcacttc cctagagaaa 2400
cacaagctgt tcatctcagg cctgcctttc tcctgtacta aagaggaact agaagaaatc 2460
tgtaaggctc atggcaccgt gaaggacctc aggctggtca ccaaccgggc tggcaaacca 2520
aagggcctgg cctacgtgga gtatgaaaat gaatcccagg cgtcgcaggc tgtgatgaag 2580
atggacggca tgactatcaa agagaacatc atcaaagtgg caatcagcaa ccctcctcag 2640
aggaaagttc cagagaagcc ggagaccagg aaggcaccag gtggccccat gcttttgccg 2700
cagacatacg gagcgacttt tggaggccca gtgggcaggc caggcagggc gggtacggag 2760
cctcccaggc tggggcagtg ggcatgggca ggggctgtgg ctgaagacct cgcccgccca 2820
ctgcagaccc caggggactc tcacaccgca gctgccatgg ccaccaataa ggagcgactc 2880
tttgcggctg gtgccctggg gcctggatct ggctacccag gggcaggttt ccccttcgcc 2940
ttcccagggg cactcagggg gtctccgcct ttcgagatgc tgagccctag cttccggggc 3000
ctgggccagc ctgacctccc caaggagatg gcctctctgt cggtggagac acagagcacc 3060
agctcagagg agatggtgcc cagctcgccc tcgccccctc cgcctcctcg ggtctacaag 3120
ccatgcttcg tgtgcaatga caagtcctct ggctaccact atggggtcag ctcttgtgaa 3180
ggctgcaagg gcttctttcg ccgaagcatc cagaagaaca tggtgtacac gtgtcaccgc 3240
gacaaaaact gtatcatcaa caaggtgacc aggaatcgct gccagtactg ccggctacag 3300
aagtgcttcg aagtgggcat gtccaaggaa gctgtgcgaa atgaccggaa caagaagaag 3360
aaagaggtga aggaagaagg gtcacctgac agctatgagc tgagccctca gttagaagag 3420
ctcatcacca aggtcagcaa agcccatcag gagactttcc cctcgctctg ccagctgggc 3480
aagtatacca cgaactccag tgcagaccac cgcgtgcagc tggatctggg gctgtgggac 3540
aagttcagtg agctggctac caagtgcatc atcaagatcg tggagtttgc caagcggttg 3600
cctggcttta cagggctcag cattgctgac cagatcactc tgctcaaagc tgcctgccta 3660
gatatcctga tgctgcgtat ctgcacaagg tacaccccag agcaggacac catgaccttc 3720
tccgacgggc tgaccctgaa ccggacccag atgcacaatg ccggcttcgg gcccctcaca 3780
gaccttgtct ttgcctttgc tgggcagctc ctgcccctgg agatggatga caccgagaca 3840
gggctgctca gcgccatctg cctcatctgc ggagaccgca tggacctgga ggagcccgaa 3900
aaagtggaca agctgcagga gccactgctg gaagccctga ggctgtacgc ccggcgccgg 3960
cggcccagcc agccctacat gttcccaagg atgctaatga aaatcaccga cctccggggc 4020
atcagcacta agggagctga aagggccatt actctgaaga tggagattcc aggcccgatg 4080
cctcccttaa tccgagagat gctggagaac cctgaaatgt ttgaggatga ctcctcgcag 4140
cctggtcccc accccaatgc ctctagcgag gatgaggttc ctgggggcca gggcaaaggg 4200
ggcctgaagt ccccagcctg a 4221
<210> 2
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
tcaaagtggc aatcagcaac c 21
<210> 3
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
agcctgggag gctccgtacc 20
<210> 4
<211> 191
<212> DNA
<213> 智人(Homo sapiens)
<400> 4
ggcctggcct acgtggagta tgaaaatgaa tcccaggcgt cgcaggctgt gatgaagatg 60
gacggcatga ctatcaaaga gaacatcatc aaagtggcaa tcagcaaccc tcctcagagg 120
aaagttccag agaagccgga gaccaggaag gcaccaggtg gccccatgct tttgccgcag 180
acatacggag c 191
<210> 5
<211> 326
<212> DNA
<213> 智人(Homo sapiens)
<400> 5
gacttttgga ggcccagtgg gcaggccagg cagggcgggt acggagcctc ccaggctggg 60
gcagtgggca tgggcagggg ctgtggctga agacctcgcc cgcccactgc agaccccagg 120
ggactctcac accgcagctg ccatggccac caataaggag cgactctttg cggctggtgc 180
cctggggcct ggatctggct acccaggggc aggtttcccc ttcgccttcc caggggcact 240
cagggggtct ccgcctttcg agatgctgag ccctagcttc cggggcctgg gccagcctga 300
cctccccaag gagatggcct ctctgt 326
<210> 6
<211> 160
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
tcaaagtggc aatcagcaac cctcctcaga ggaaagttcc agagaagcca gagaccagga 60
aggcaccagg tggccccatg cttttgccgc agacatacgg agcgactttt ggaggcccag 120
tgggcaggcc aggcagggcg ggtacggagc ctcccaggct 160
Claims (10)
1.一种AML相关的融合基因，其特征在于，由SART3外显子18及其上游序列和RARG外显子3及其下游序列，通过SART3外显子18和RARG外显子3直接融合形成，其序列见SEQ IDNO.1所示。
2.检测权利要求1所述的AML相关的融合基因的试剂在制备AML辅助诊断制剂中的应用。
3.根据权利要求1所述的应用，其特征在于，辅助鉴别经典APL。
4.根据权利要求1所述的应用，其特征在于，检测所述的AML相关的融合基因的试剂包括PCR检测试剂。
5.根据权利要求4所述的应用，其特征在于，所述的PCR检测试剂中上游引物以SART3外显子18为靶基因设计，下游引物以RARG外显子3为靶基因设计。
6.根据权利要求5所述的应用，其特征在于，所述引物序列如SEQ ID NO.2和SEQ IDNO.3所示。
7.用于扩增权利要求1所述的AML相关的融合基因的引物，其特征在于，所述引物序列如SEQ ID NO.2和SEQ ID NO.3所示。
8.一种AML诊断试剂盒，其特征在于，包括检测所述融合基因的试剂。
9.根据权利要求1所述的应用，其特征在于，辅助鉴别经典APL。
10.根据权利要求8所述的试剂盒，其特征在于，检测所述的AML相关的融合基因的试剂包括PCR检测试剂；进一步地，所述的PCR检测试剂中上游引物以SART3外显子18为靶基因设计，下游引物以RARG外显子3为靶基因设计；更进一步地，所述引物序列如SEQ ID NO.2和SEQ ID NO.3所示。
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| CN117165609A (zh) * | 2023-09-06 | 2023-12-05 | 上海新培晶医学检验所有限公司 | 一种cbfa2t3-nat16融合基因及其检测引物和应用 |
| CN117165610A (zh) * | 2023-10-31 | 2023-12-05 | 南昌大学第一附属医院 | 一种rara相关变异型apl的融合基因及其扩增引物 |
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| CN111549043A (zh) * | 2020-06-09 | 2020-08-18 | 苏州大学附属第一医院 | 一种rara相关变异型apl的融合基因及其检测引物和应用 |
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Non-Patent Citations (2)
| Title |
|---|
| JI LI等: "A novel SART3::RARG fusion gene in acute myeloid leukemia with acute promyelocytic leukemia phenotype and differentiation escape to retinoic acid", 《HAEMATOLOGICA》, vol. 108, no. 2, pages 627 - 632 * |
| 张静等: "急性早幼粒细胞白血病相关融合基因的研究进展", 《医学综述》, vol. 25, no. 10, pages 1894 - 1899 * |
Cited By (2)
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|---|---|---|---|---|
| CN117165609A (zh) * | 2023-09-06 | 2023-12-05 | 上海新培晶医学检验所有限公司 | 一种cbfa2t3-nat16融合基因及其检测引物和应用 |
| CN117165610A (zh) * | 2023-10-31 | 2023-12-05 | 南昌大学第一附属医院 | 一种rara相关变异型apl的融合基因及其扩增引物 |
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