No abstract available.
Aged, 80 and over ; Base Sequence ; Bone Marrow/pathology ; DNA/chemistry/metabolism ; Female ; Fusion Proteins, bcr-abl/*genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/diagnosis/*genetics ; Multiplex Polymerase Chain Reaction ; Protein Isoforms/genetics ; Sequence Analysis, DNA

No abstract available.
Aged ; Base Sequence ; Bone Marrow/metabolism/pathology ; DNA Mutational Analysis ; Female ; Fusion Proteins, bcr-abl/*genetics ; Gene Duplication ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myeloid, Acute/diagnosis/*genetics ; Multiplex Polymerase Chain Reaction ; Mutation ; Nuclear Proteins/*genetics ; Philadelphia Chromosome ; fms-Like Tyrosine Kinase 3/*genetics

No abstract available.
Antineoplastic Agents/therapeutic use ; Base Sequence ; DNA/chemistry/genetics/metabolism ; DNA Mutational Analysis ; Fusion Proteins, bcr-abl/*genetics ; Genotype ; Humans ; Imatinib Mesylate/therapeutic use ; Karyotyping ; Male ; Multiplex Polymerase Chain Reaction ; Mutation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/*diagnosis/drug therapy/*genetics

OBJECTIVETo investigate the accuracy and consistency of the detection of BCR-ABL tyrosine kinase domain point mutation among different laboratories.

METHODSEvery one of 6 laboratories prepared 10 cDNA samples from tyrosine kinase inhibitors resistant BCR-ABL (P210 or P190) positive patients'bone marrow or peripheral blood. Each cDNA sample was divided into 6 aliquots and delivered to the laboratories. All 6 laboratories tested BCR-ABL point mutations of 60 samples according to their own protocols. Peking University People's Hospital analyzed the comparison results based on both the reports and sequencing chromatogram from all laboratories.

RESULTSAll laboratories reported the same nucleotide and corresponding amino acid mutations in 37 samples (61.7%）. Of 60 samples, 53 had confirmed mutation types, and a total of 23 types were included; 1 had no mutation; mutation types of 6 samples could not be determined because of the big differences among chromatograms from different laboratories. Low percentages of mutants were significantly related to results inconsistency (P=0.008). Inconsistent result of one sample was caused by the unique chromatogram of the mutant L248V, and one by the non-coverage amplification of PCR product from different laboratories. Amplification was failed in 3 samples. Testing or sequencing mistakes occurred in 7 samples. The differences in the mutant percentages among laboratories were less than 20% in the 80.6% of samples with confirmed results. Low internal control gene copies (ABL<10 000) were significantly related to both failed amplification and big differences among chromatograms from different laboratories (P=0.005 and <0.001, respectively).

CONCLUSIONProblems in the clinical routine detection of BCR-ABL point mutation could be exposed and improvement could be achieved by sample exchange and comparison. Low percentage of mutant is the main reason which causes the discrepancy of BCR-ABL point mutation results among different laboratories.

Bone Marrow ; DNA Mutational Analysis ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Point Mutation ; Polymerase Chain Reaction

OBJECTIVETo identify IKZF1 gene copy number abnormalities in BCR/ABL-negative B-lineage acute lymphoblastic leukemia (B-ALL) in children, and to investigate the association between such abnormalities and prognosis.

METHODSMultiplex ligation-dependent probe amplification (MLPA) was applied to detect IKZF1 gene copy number abnormalities in 180 children diagnosed with BCR/ABL-negative B-ALL. These children were classified into IKZF1 deletion group and IKZF1 normal group according to the presence or absence of IKZF1 gene deletion. The association between IKZF1 copy number abnormalities and prognosis of children with BCR/ABL-negative B-ALL was analyzed retrospectively.

RESULTSAmong 180 children, 27 (15.0%) had IKZF1 deletion; among the 27 children, 4 had complete deletions of 8 exons of IKZF1 gene, 17 had deletion of exon 1, 3 had deletions of exons 4-7, and 3 children had deletions of exons 2-7. Compared with those in the IKZF1 normal group, children in the IKZF1 deletion group had higher white blood cell (WBC) count and percentage of individuals with high risk of minimal residual disease at the first visit. IKZF1 deletions often occurred in BCR/ABL-negative children with no special fusion gene abnormalities. They were frequently accompanied by abnormalities in chromosomes 11, 8, 5, 7, and 21. The analysis with Kaplan-Meier method showed that disease-free survival (DFS) in the IKZF1 deletion group was significantly lower than that in the IKZF1 normal group (0.740 ± 0.096 vs 0.905 ± 0.034; P=0.002). Cox analysis showed that after exclusion of sex, age, initial WBC count, cerebrospinal fluid state at the first visit, prednisone response, and chromosome karyotype, IKZF1 deletion still affected the children's DFS (P<0.05).

CONCLUSIONSSome children with BCR/ABL-negative B-ALL have IKZF1 deletion, and IKZF1 deletion is an independent risk factor for DFS in children with BCR/ABL-negative B-ALL.

Adolescent ; Child ; Child, Preschool ; Female ; Fusion Proteins, bcr-abl ; analysis ; Gene Dosage ; Humans ; Ikaros Transcription Factor ; genetics ; Infant ; Male ; Multiplex Polymerase Chain Reaction ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; mortality ; Prognosis

The genetic variant rs16754 of Wilms tumor gene 1 (WT1) has recently been described as an independent prognostic factor in AML patients. It is of great interest to test whether WT1 single nucleotide polymorphism can be used as a molecular marker in other types of cancer, to improve risk and treatment stratification. We performed sequencing analysis of exons 7 and 9 of WT1, which are known mutational hotspots, in a total of 73 patients with BCR-ABL1-negative myeloproliferative neoplasm (MPN) and 93 healthy controls. No previously reported WT1 mutations were identified in the present study. In Korean patients with BCR-ABL1-negative MPN, WT1 genetic variant rs16754 had no significant impact on clinical outcomes. We observed a significant difference in the allelic frequencies of WT1 rs16754 in Koreans between BCR-ABL1-negative MPN cases and healthy controls. Individuals carrying variant G alleles of WT1 rs16754 showed a relatively low prevalence of BCR-ABL1-negative MPN, compared with those carrying wild A alleles of WT1 rs16754 (Hazard ratio 0.10-0.65, P<0.05). Therefore, possession of the variant G allele of WT1 rs16754 may reduce the risk of developing BCR-ABL1-negative MPN.
Adult ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group/*genetics ; Case-Control Studies ; Exons ; Female ; Fusion Proteins, bcr-abl/genetics ; Gene Frequency ; Genotype ; Humans ; Leukemia, Myeloid, Acute/pathology ; Male ; Middle Aged ; Myeloproliferative Disorders/*genetics/pathology ; Polymorphism, Single Nucleotide ; Prognosis ; Proportional Hazards Models ; Republic of Korea ; Risk ; Sequence Analysis, DNA ; WT1 Proteins/*genetics ; Young Adult

No abstract available.
Adult ; Biomarkers, Tumor/genetics ; Bone Marrow/pathology ; Calreticulin/genetics ; Erythropoietin/blood ; Female ; Fusion Proteins, bcr-abl/*genetics ; Hematocrit ; Hemoglobins/analysis ; Humans ; Janus Kinase 2/genetics ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders/*diagnosis/genetics ; Polycythemia Vera/*diagnosis/genetics ; Receptors, Thrombopoietin/genetics ; Thrombocythemia, Essential/diagnosis ; World Health Organization

No abstract available.
Aged ; Base Sequence ; Bone Marrow/metabolism/pathology ; Chromosome Aberrations ; DNA/chemistry/genetics/metabolism ; Fusion Proteins, bcr-abl/*genetics ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Male ; Myelodysplastic Syndromes/diagnosis/*genetics ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo evaluate the efficiency of one-step multiplex RT-PCR for identifying four common fusion transcripts (TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL) in children with acute lymphoblastic leukemia (ALL).

METHODSTotal RNA was extracted from bone marrow samples of 76 children who were newly diagnosed with ALL between January 2003 and December 2010. These RNAs were analyzed for TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL by one-step multiplex RT-PCR or common nested-multiplex PCR. The PCR products were confirmed by DNA sequencing.

RESULTSTEL/AML1 was found in 12 cases (the length of products was 298 bp in 9 cases and 259 bp in 3 cases), E2A/PBX1 was found in 3 cases (the length of products was 373 bp), BCR/ABL was found in 1 case (the length of products was 2 124 bp), and MLL/AF4 was found in 7 cases (the length of products was 427 bp in 1 case and 673 bp in 6 cases) using one-step multiplex RT-PCR combined with DNA sequencing. The results were consistent with those using common nested-multiplex PCR.

CONCLUSIONSOne-step multiplex RT-PCR may be another alternative for detection of common fusion transcripts in children with ALL.

Child ; Child, Preschool ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Infant ; Male ; Multiplex Polymerase Chain Reaction ; methods ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

OBJECTIVECompare the correlation and the concordance of simultaneous bone marrow cytogenetic and peripheral blood molecular responses in patients with chronic myeloid leukemia (CML) during tyrosine kinase inhibitor (TKI) treatment.

METHODSA total of 1 030 pairs of simultaneous bone marrow and peripheral blood samples from 419 patients with CML during TKI treatment were analyzed with standard G-banding techniques and real-time quantitative reverse transcriptase PCR (Q-PCR).

RESULTSThe Spearman correlation coefficient for the paired analysis of the percentage of Ph- positive metaphases versus Q-PCR (BCR-ABL/ABL) values was 0.655 (n=1 030, P<0.01). There was a significant difference in terms of BCR-ABL values among all three cytogenetic response groups of no partial cytogenetic (no PCyR), partial cytogenetic (PCyR) and complete cytogenetic responses(CCyR) (P<0.01). 93.8% of the patients in CCyR had BCR-ABL≤1% [International Scale (IS)], and 97.5% of those with BCR-ABL≤0.1% (IS) were in CCyR. There was good concordance of 86.2% (888 of 1 030 samples) when BCR-ABL values according to cutoffs of BCR-ABL>10%(IS), ≤10%->1%(IS)and ≤1%(IS) were coupled with cytogenetic responses including no PCyR, PCyR and CCyR. Furthermore, 497 pairs of samples from 279 patients with newly diagnosed CML in chronic phase during the first year on TKI as first-line therapy were analyzed. Concordances between major cytogenetic response and BCR-ABL≤10%(IS) at 3 months, CCyR and BCR-ABL≤1%(IS) at 6 months and 12 months were 89%, 83.5% and 92.1%, respectively.

CONCLUSIONThere were significant correlation and concordance between cytogenetic and molecular responses, and some differences in assessment of early responses between using cytogenetic and molecular analyses in CML patients during TKI treatment.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; cytology ; drug effects ; Child ; Cytogenetic Analysis ; Female ; Fusion Proteins, bcr-abl ; antagonists & inhibitors ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; Male ; Middle Aged ; Philadelphia Chromosome ; Protein Kinase Inhibitors ; therapeutic use ; Treatment Outcome ; Young Adult