一般情况下RQ-PCR检测NPM1的敏感性高于FCM,但也有部分患者出现NPM1-FCM+,可能与NPM1探针针对特殊类型突变的敏感性、疾病过程中的克隆演变以及标本的取材等原因有关。而此时的FCM+对复发的预测也有意义。 综上,在AML化疗过程中监测MRD,FCM+或NPM1+均对复发...
1、实时荧光定量PCR检测NPMl一mu认mRNA表达量的方法构建:采用相 对定量法,应用TaqMan探针技术,以管家基因ABL为内参,同时设置正常对照 和无模板对照;提取待测标本总RNA,测定其核酸浓度,进行10倍系列稀释后作 为反应模板,设定其拷贝数为101、102、103、104、105、106构建标准曲线;反应 ...
After being washed three times with low-salt wash buffer and one time with high-salt wash buffer, immunoprecipitated chromatin DNA was eluted and quantified using real-time quantitative PCR. Sequences of primers for ChIP Q-PCR were listed in Table S4. Statistical analysis Statistical tests and ...
Methods NPM1 mutation(including A,B,D mutation type) was detected in 206 patients with newly diagnosed AML by real-time quantitative RT-PCR. Results The incidence of NPM1 mutation was 15.5% in total AML patients and 32.5% in normal karyotypes AML patients.The characteristics of 174 NPM1 ...
(TaKaRa Bio Inc., Kusatsu, Japan), and reversely transcribed into cDNA using a Reverse Transcriptase Kit (Invitrogen, Carlsbad, CA, USA), followed by RT-qPCR using SYBR Premix Ex Taq II (Takara, Shiga, Japan) in an ABI-7300 Real-Time PCR system (Applied Biosystems, Foster City, CA, ...
(q22) or otherwise go unrecognized. FISH or RT-PCR studies are therefore suggested when the presence ofCBFβ/MYH11is suggested by the morphologic findings but classical cytogenetic studies have been negative or uninformative. Quantitative real-time RT-PCR approaches have been employed for monitoring ...
AML karyotype analysis was normal (46,XY[20]) with BCR-ABL1 transcripts undetectable by both real-time quantitative PCR and nested PCR. Further PCR analysis identified NPM1mut type B without FLT3-ITD. Interestingly, subsequent testing demonstrated that the NPM1 mutation was present at low levels...
RealTime PCR (RT-PCR) was performed using the BioRad CFX96 Touch using TaqMan universal master mix and FAM probes. For HOXA9 qPCR following ATRA/ATO, SYBR Green PCR master mix and custom primers were used instead of FAM probes. The mRNA levels of test genes were normalized to GAPDH ...
Assessment of measurable residual disease (MRD) by real time quantitative polymerase chain reaction (RT-qPCR) forNPM1mutant transcripts [11] can be combined with the ELN risk stratification to inform therapeutic decisions, e.g. helping to select patients who may benefit from allogeneic haematopoietic...
Multiparameter flow cytometry (MFC) and real-time quantitative polymerase chain reaction (RT-PCR) were used for monitoring MRD. Ultimately, 106 patients were included in the long-term follow-up period. Patients with high NPM1 VAF (≥ 42.43%) had poorer 2-year relapse-free survival (RFS) ...