CLOSE X
CLOSE X
• JAK2 V617F is located on chromosome 9, exon 14
• Quantitative PCR can identify the V617F (1849G>T) mutation in the JAK2 gene
• Present in the majority of patients with polycythemia vera, and in a subset of patients with essential thrombocythemia, primary myelofibrosis, and in rare cases, other myeloid malignancies (see image)
• Genoptix offers serial reporting by PCR, capable of detecting JAK2 mutations with a sensitivity level of 1%
JAK2
< BACK
VIEW REPORT >
CLOSE X
CLINICAL DATA:
56-year-old male. Rule out CML. Whole blood submitted for evaluation.
RESULTS:
Results of JAK2 testing by Invader Plus® were POSITIVE, indicating the V617F (1849G>T) mutation was pres ent in the JAK2 gene.
This mutation, which results in the constitutive action of the JAK2 tyrosine kinase, is present in the majority of patients with polycythemia vera, and in a subset of patients with essential thrombocythemia and chronic idiopathic myelofibrosis. Constitutive action of the JAK2 tyrosine kinase results in myeloproliferation in these disorders.
Nucleic acid studies do not constitute a definitive diagnostic test for all chronic myeloproliferative neoplasms but, together with other clinicopathologic findings, can support their diagnosis. Specifically, as mixed myeloproliferative syndromes are not uncommon, it is recommended that a careful correlation with bone marrow histology be performed.
Method:
MPL W515 mutation analysis includes isolation of DNA, target gene amplification via the polymerase chain reaction (PCR), and a Cleavase-based allele-specific signal amplification to determine the presence of the W515L (TGG>TTG) and W515K (TGG>AAG) genotypes. As assessed by positive DNA dilution experiments, this assay can detect mutant DNA when present at levels as low as 5%.
Technical Note:
A majority of patients with chronic myeloproliferative neoplasms will be negative for the MPL W515 mutation. Since the mutation typically is a somatic mutation, peripheral blood specimens with predominantly normal lymphocytes, or other cells not carrying the mutation, may have mutation levels below the limit of detection for this assay. Genetic testing errors may result from trace contamination of PCR reactions and from rare genetic variants that interfere with analysis.
56-year-old male. Rule out CML. Whole blood submitted for evaluation.
RESULTS:
Results of JAK2 testing by Invader Plus® were POSITIVE, indicating the V617F (1849G>T) mutation was pres ent in the JAK2 gene.
This mutation, which results in the constitutive action of the JAK2 tyrosine kinase, is present in the majority of patients with polycythemia vera, and in a subset of patients with essential thrombocythemia and chronic idiopathic myelofibrosis. Constitutive action of the JAK2 tyrosine kinase results in myeloproliferation in these disorders.
Nucleic acid studies do not constitute a definitive diagnostic test for all chronic myeloproliferative neoplasms but, together with other clinicopathologic findings, can support their diagnosis. Specifically, as mixed myeloproliferative syndromes are not uncommon, it is recommended that a careful correlation with bone marrow histology be performed.
Method:
MPL W515 mutation analysis includes isolation of DNA, target gene amplification via the polymerase chain reaction (PCR), and a Cleavase-based allele-specific signal amplification to determine the presence of the W515L (TGG>TTG) and W515K (TGG>AAG) genotypes. As assessed by positive DNA dilution experiments, this assay can detect mutant DNA when present at levels as low as 5%.
Technical Note:
A majority of patients with chronic myeloproliferative neoplasms will be negative for the MPL W515 mutation. Since the mutation typically is a somatic mutation, peripheral blood specimens with predominantly normal lymphocytes, or other cells not carrying the mutation, may have mutation levels below the limit of detection for this assay. Genetic testing errors may result from trace contamination of PCR reactions and from rare genetic variants that interfere with analysis.
Quantitative JAK2 Mutation Analysis
V617F (1849G>T) by Invader Plus®
V617F (1849G>T) by Invader Plus®
< BACK
< VIEW MARKER
References:
1. Pardanani AD, Levine RL, Lasho T, Pikman Y, Mesa RA, Wadleigh M, Steensma DP, Elliot MA, Wolanskyj AP, Hogan WJ, McClure RF, Litzow MR, Gilliland DG, and Tefferi A. MPL515 mutations in myeloproliferative and other myeloid disorders: a study of 1182 patients. Blood. 2006 Nov 15;108:3472-3476.
2. Duensing S, Duensing A, Meran JG, Kreft A, Busche G, Ganser A, Georgii A. Molecular detection of c-mpl thrombopoietin receptor gene expression in chronic myeloproliferative disorders. Mol Pathol. 1999 Jun;52(3):146-50.
1. Pardanani AD, Levine RL, Lasho T, Pikman Y, Mesa RA, Wadleigh M, Steensma DP, Elliot MA, Wolanskyj AP, Hogan WJ, McClure RF, Litzow MR, Gilliland DG, and Tefferi A. MPL515 mutations in myeloproliferative and other myeloid disorders: a study of 1182 patients. Blood. 2006 Nov 15;108:3472-3476.
2. Duensing S, Duensing A, Meran JG, Kreft A, Busche G, Ganser A, Georgii A. Molecular detection of c-mpl thrombopoietin receptor gene expression in chronic myeloproliferative disorders. Mol Pathol. 1999 Jun;52(3):146-50.
CLOSE X
CLOSE X
• IgVH mutation analysis by PCR is part of the prognostic profile for CLL
- IgVH gene mutation >2% - positive prognostic indicator1
- IgVH gene mutation ≤2% - negative prognostic indicator1
Kaplan-Meier survival curve comparing CLL patients with mutated and unmutated VH genes.
- IgVH gene mutation >2% - positive prognostic indicator1
- IgVH gene mutation ≤2% - negative prognostic indicator1
Kaplan-Meier survival curve comparing CLL patients with mutated and unmutated VH genes.
IgVH
< BACK
VIEW REPORT >
Reference: 1. Hamblin T, Davis Z, Gardiner A, et al. Unmutated Ig VH genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood.(1999)94:1848-1854.
CLOSE X
Kaplan-Meier survival curve comparing CLL patients with mutated and unmutated VH genes. Median survival for unmutated CLL: 117 months; median survival for mutated CLL: 293 months. The difference is significant at the P = .001 level (log-rank test).
CLINICAL DATA:
58-year-old male with anemia. Bone marrow submitted for evaluation. Accompanying CBC report, dated 3/12/10, indicates WBC 7.94 K/uL, RBC 3.78 M/uL, Hgb 11.8 g/dL, HCT 33.4%, MCV 88.3 fL, MCH 31.1 pg, MCHC 35.2 g/dL, RDW 16.0%, platelets 172 K/uL with a differential count of neutrophils 50.9%, lymphocytes 43.3%, monocytes 3.82%, eosinophils 0.977%, basophils 0.990%.
RESULTS:
PCR amplification of the IgH gene variable (V) region detects a monoclonal population of B-cells. DNA sequencing of the amplified product is performed and is compared to the germline (unmutated) consensus sequence. This patient’s sequence is 0% different from germline. Since there is 2% or less divergence, it is considered UNMUTATED and thus a POORER PROGNOSIS for CLL.
Interpretation:
Somatic hypermutation of the IgH V region occurs naturally during B-cell maturation in lymphoid follicles. In CLL patients, a clonal B-cell population with a hypermutated IgH V region (greater than 2% of DNA nucleotides different from the reference germline sequence) suggests a more mature malignancy and correlates with a more favorable prognosis and a significantly longer median survival time than those with unmutated (less than or equal to 2% difference) IgH V region (1,2,3,4).
Method:
Patient RNA is isolated and subjected to RT-PCR using two independent multiplex reactions. Each reaction contains consensus oligonucleotide primer sets specific for the upstream leader (L) or framework 1 (FR1) regions, and the downstream joining (J) region of the IgH gene on chromosome 14. PCR products are resolved using high resolution capillary electrophoresis to confirm clonality. The DNA nucleotide sequence of the IgH V region is derived and compared to standard germline sequence families contained in the National Center for Biotechnology Information IgBLAST database. The number of mutations in the patient’s IgH V region clone is expressed as a percentage in comparison to the closest matching IgH family sequence.
The limit of detection determined by RNA mixing tests is 10% clonal RNA in a polyclonal RNA background. This means that in patient specimens containing less than 10% clonal B cells, we may be unable to obtain adequate PCR product or sequencing signal. PCR may fail due to inadequate oligonucleotide primer hybridization caused by unpredicted mutations which may occur in the targeted primer binding sites. Results of molecular testing should be interpreted in the context of clinical, histological and immunophenotypic data.
58-year-old male with anemia. Bone marrow submitted for evaluation. Accompanying CBC report, dated 3/12/10, indicates WBC 7.94 K/uL, RBC 3.78 M/uL, Hgb 11.8 g/dL, HCT 33.4%, MCV 88.3 fL, MCH 31.1 pg, MCHC 35.2 g/dL, RDW 16.0%, platelets 172 K/uL with a differential count of neutrophils 50.9%, lymphocytes 43.3%, monocytes 3.82%, eosinophils 0.977%, basophils 0.990%.
RESULTS:
PCR amplification of the IgH gene variable (V) region detects a monoclonal population of B-cells. DNA sequencing of the amplified product is performed and is compared to the germline (unmutated) consensus sequence. This patient’s sequence is 0% different from germline. Since there is 2% or less divergence, it is considered UNMUTATED and thus a POORER PROGNOSIS for CLL.
Interpretation:
Somatic hypermutation of the IgH V region occurs naturally during B-cell maturation in lymphoid follicles. In CLL patients, a clonal B-cell population with a hypermutated IgH V region (greater than 2% of DNA nucleotides different from the reference germline sequence) suggests a more mature malignancy and correlates with a more favorable prognosis and a significantly longer median survival time than those with unmutated (less than or equal to 2% difference) IgH V region (1,2,3,4).
Method:
Patient RNA is isolated and subjected to RT-PCR using two independent multiplex reactions. Each reaction contains consensus oligonucleotide primer sets specific for the upstream leader (L) or framework 1 (FR1) regions, and the downstream joining (J) region of the IgH gene on chromosome 14. PCR products are resolved using high resolution capillary electrophoresis to confirm clonality. The DNA nucleotide sequence of the IgH V region is derived and compared to standard germline sequence families contained in the National Center for Biotechnology Information IgBLAST database. The number of mutations in the patient’s IgH V region clone is expressed as a percentage in comparison to the closest matching IgH family sequence.
The limit of detection determined by RNA mixing tests is 10% clonal RNA in a polyclonal RNA background. This means that in patient specimens containing less than 10% clonal B cells, we may be unable to obtain adequate PCR product or sequencing signal. PCR may fail due to inadequate oligonucleotide primer hybridization caused by unpredicted mutations which may occur in the targeted primer binding sites. Results of molecular testing should be interpreted in the context of clinical, histological and immunophenotypic data.
IgVH Hypermutation Analysis
< BACK
< VIEW MARKER
References
1. Hamblin et al. Unmutated IgV(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood (1999) 94, 1848-1854.
2. Damle et al IgV gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood (1999) 94, 1840-1847.
3. Matthews et al. Routine Analysis of IgVH mutational status in CLL patients using BIOMED-2 standardized primers and protocols. Leukemia & Lymphoma, (2004) 45(9), 1899-1904.
4. Krober et al V(H) mutation status, CD38 expression level, genomic aberrations, and survival in chronic lymphocytic leukaemia. Blood (2002) 100, 1410-1416.
1. Hamblin et al. Unmutated IgV(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood (1999) 94, 1848-1854.
2. Damle et al IgV gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood (1999) 94, 1840-1847.
3. Matthews et al. Routine Analysis of IgVH mutational status in CLL patients using BIOMED-2 standardized primers and protocols. Leukemia & Lymphoma, (2004) 45(9), 1899-1904.
4. Krober et al V(H) mutation status, CD38 expression level, genomic aberrations, and survival in chronic lymphocytic leukaemia. Blood (2002) 100, 1410-1416.
CLOSE X
CLOSE X
• t(15;17) is a recurrent genetic abnormality found in patients with acute promyelocytic leukemia (APL)
- Can be detected by FISH or PCR
- Can be detected by FISH or PCR
• NCCN guidelines recommends PCR as methodology for monitoring molecular remission of APL post treatment intervention
• Genoptix notifies physicians of acute cases within 24 hours or less, and all acute cases are confirmed by a second Genoptix hematopathologist
PML-RARA
< BACK
VIEW REPORT >
References: NCCN Practice Guidelines in Oncology. Acute Myeloid Leukemia. V.2.2011.
CLOSE X
CLINICAL DATA:
55-year-old female with acute promyelocytic leukemia, status post induction with 7+3+ATRA. Other treatment includes Daunorubicin Ara-C. Previous Compass Report reveals peripheral blood with acute promyelocytic leukemia with t(15;17), PML-RARA (collected 8/4/11, 200242978). Bone marrow submitted for evaluation. Accompanying CBC report, dated 10/28/11, indicates WBC 5.5 K/uL, RBC 3.36 M/uL, Hgb 10.6 g/dL, HCT 29.5%, MCV 87.8 fL, MCH 31.6 pg, MCHC 36.0 g/dL, RDW 13.3%, platelets 149 K/uL with a differential count of neutrophils 77%, lymphocytes 16%, monocytes 5%, eosinophils 2%.
Comments:
A t(15;17) translocation is not detected, indicating a NEGATIVE result. This result does not entirely exclude the possibility of a t(15;17) translocation present in the sample below the quantifiable detection limit of this test.
Notes:
This test is designed to detect t(15;17) PML-RARA, a recurrent genetic abnormality found in a subset of patients with acute myeloid leukemia. This test detects all three gene fusion patterns: type A (short, S-form, bcr-3), type B (long, L-form, bcr-1), and type B variant (variable, V-form, bcr-2).
METHODOLOGY:
Patient RNA is isolated, reverse transcribed into cDNA, and subjected to PCR amplification (RT-PCR) using primers specific for the PML and RARA genes. PML-RARA and ABL (control) transcripts are quantified by comparison to a standard curve run with each assay. Results are reported as a normalized ratio of PML-RARA transcripts to ABL transcripts present in the sample. Positive controls, negative controls, and controls for sample quality are performed in parallel.
LIMITATIONS:
Translocations involving other genes or gene partners will not be detected.
LIMIT OF DETECTION:
1 in 10,000 cells. Results of this test must always be interpreted in the context of morphologic and other relevant data, and should not be used alone for a diagnosis of malignancy. This test is not intended to detect minimal residual disease.
Note: Specimen is greater than 48 hours old upon receipt at testing laboratory. Sample is extracted past stability time limits. Samples submitted for RNA testing should be extracted within 48 hours. Test results should be interpreted with caution.
55-year-old female with acute promyelocytic leukemia, status post induction with 7+3+ATRA. Other treatment includes Daunorubicin Ara-C. Previous Compass Report reveals peripheral blood with acute promyelocytic leukemia with t(15;17), PML-RARA (collected 8/4/11, 200242978). Bone marrow submitted for evaluation. Accompanying CBC report, dated 10/28/11, indicates WBC 5.5 K/uL, RBC 3.36 M/uL, Hgb 10.6 g/dL, HCT 29.5%, MCV 87.8 fL, MCH 31.6 pg, MCHC 36.0 g/dL, RDW 13.3%, platelets 149 K/uL with a differential count of neutrophils 77%, lymphocytes 16%, monocytes 5%, eosinophils 2%.
Comments:
A t(15;17) translocation is not detected, indicating a NEGATIVE result. This result does not entirely exclude the possibility of a t(15;17) translocation present in the sample below the quantifiable detection limit of this test.
Notes:
This test is designed to detect t(15;17) PML-RARA, a recurrent genetic abnormality found in a subset of patients with acute myeloid leukemia. This test detects all three gene fusion patterns: type A (short, S-form, bcr-3), type B (long, L-form, bcr-1), and type B variant (variable, V-form, bcr-2).
METHODOLOGY:
Patient RNA is isolated, reverse transcribed into cDNA, and subjected to PCR amplification (RT-PCR) using primers specific for the PML and RARA genes. PML-RARA and ABL (control) transcripts are quantified by comparison to a standard curve run with each assay. Results are reported as a normalized ratio of PML-RARA transcripts to ABL transcripts present in the sample. Positive controls, negative controls, and controls for sample quality are performed in parallel.
LIMITATIONS:
Translocations involving other genes or gene partners will not be detected.
LIMIT OF DETECTION:
1 in 10,000 cells. Results of this test must always be interpreted in the context of morphologic and other relevant data, and should not be used alone for a diagnosis of malignancy. This test is not intended to detect minimal residual disease.
Note: Specimen is greater than 48 hours old upon receipt at testing laboratory. Sample is extracted past stability time limits. Samples submitted for RNA testing should be extracted within 48 hours. Test results should be interpreted with caution.
PML/RARA, t(15;17) by PCR, Quantitative
< BACK
< VIEW MARKER
CLOSE X
CLOSE X
• cKIT or CD117 is a proto-oncogene; over expression or mutations of this protein can lead to cancer1
• Prognosis is unfavorable with detected mutation2,3
• Active in several liquid and solid tumor types; therefore results of this test should always be interpreted in the context of morphologic and other relevant data, and should not be used alone for a diagnosis of malignancy
cKIT
< BACK
VIEW REPORT >
References:
1.Edling CE, Hallberg B (2007). c-Kit--a hematopoietic cell essential receptor tyrosine kinase. Int. J. Biochem. Cell Biol. 39 (11): 1995–8
2.Schlenk RF. Mutations and treatment outcomes in cytogenetically normal acute myeloid leukemia. N Engl J Med. 2008;358;18, 1909–1918.
3.Sanz MA. Management of acute promyelocytic leukemia: recommendations from an expert panel on behalf of the European Leukemia Net, Blood. 2009;113, 1875–1891.
1.Edling CE, Hallberg B (2007). c-Kit--a hematopoietic cell essential receptor tyrosine kinase. Int. J. Biochem. Cell Biol. 39 (11): 1995–8
2.Schlenk RF. Mutations and treatment outcomes in cytogenetically normal acute myeloid leukemia. N Engl J Med. 2008;358;18, 1909–1918.
3.Sanz MA. Management of acute promyelocytic leukemia: recommendations from an expert panel on behalf of the European Leukemia Net, Blood. 2009;113, 1875–1891.
References:
2)Schlenk RF. Mutations and treatment outcomes in cytogenetically normal acute myeloid leukemia. N Engl J Med. 2008;358;18, 1909–1918.
4)Sanz MA. Management of acute promyelocytic leukemia: recommendations from anexpert panel on behalf of the European Leukemia Net, Blood. 2009;113, 1875–1891.
2)Schlenk RF. Mutations and treatment outcomes in cytogenetically normal acute myeloid leukemia. N Engl J Med. 2008;358;18, 1909–1918.
4)Sanz MA. Management of acute promyelocytic leukemia: recommendations from anexpert panel on behalf of the European Leukemia Net, Blood. 2009;113, 1875–1891.
CLOSE X
CLINICAL DATA:
65-year-old female with persistent severe anemia and myelodysplasia. Rule out acute leukemia.
Bone marrow submitted for evaluation.
Accompanying CBC report, dated 1/31/11, indicates WBC 7.49 K/uL, RBC 2.28 M/uL, Hgb 6.7 g/dL, HCT 20.3%, MCV 88.9 fL, MCH 29.2 pg, MCHC 32.9g/dL, RDW 15.7%, platelets 161.0 K/uL with a differential count of neutrophils 46.0%, bands 21.0, lymphocytes 16.0%, eosinophils 3.0%, metamyelocytes 8%, myelocytes 6.0%, 1+ anisocytosis, 1+ poikilocytosis, few ovalocytes, immature granulocytes present, 1+ toxic granulation.
Comments:
There is evidence of the cKIT (D816V) point mutation by PCR analysis, indicating a POSITIVE result.
Notes:
Patient DNA is isolated and subjected to allele-specific PCR amplification. The reaction uses an oligonucleotide primer set specific for the exon 17 of KIT on chromosome 4, and an allele-specific primer that specifically initiates amplification from the allele containing the point mutation in codon 816. Each assay includes a positive control reaction using DNA from a plasmid that contains the KIT D816V mutation and a negative control using placental DNA. The KIT gene sequences are present in the normal human genome and serve as a control for PCR in the assay.
PCR products are analyzed by electrophoresis and UV transillumination of ethidium bromide stained gels.
Results of this test must always be interpreted in the context of morphologic and other relevant data, and should not beused alone for a diagnosis of malignancy.
The D816V mutation can be detected if tumor represents at least 0.3% of cells.
65-year-old female with persistent severe anemia and myelodysplasia. Rule out acute leukemia.
Bone marrow submitted for evaluation.
Accompanying CBC report, dated 1/31/11, indicates WBC 7.49 K/uL, RBC 2.28 M/uL, Hgb 6.7 g/dL, HCT 20.3%, MCV 88.9 fL, MCH 29.2 pg, MCHC 32.9g/dL, RDW 15.7%, platelets 161.0 K/uL with a differential count of neutrophils 46.0%, bands 21.0, lymphocytes 16.0%, eosinophils 3.0%, metamyelocytes 8%, myelocytes 6.0%, 1+ anisocytosis, 1+ poikilocytosis, few ovalocytes, immature granulocytes present, 1+ toxic granulation.
Comments:
There is evidence of the cKIT (D816V) point mutation by PCR analysis, indicating a POSITIVE result.
Notes:
Patient DNA is isolated and subjected to allele-specific PCR amplification. The reaction uses an oligonucleotide primer set specific for the exon 17 of KIT on chromosome 4, and an allele-specific primer that specifically initiates amplification from the allele containing the point mutation in codon 816. Each assay includes a positive control reaction using DNA from a plasmid that contains the KIT D816V mutation and a negative control using placental DNA. The KIT gene sequences are present in the normal human genome and serve as a control for PCR in the assay.
PCR products are analyzed by electrophoresis and UV transillumination of ethidium bromide stained gels.
Results of this test must always be interpreted in the context of morphologic and other relevant data, and should not beused alone for a diagnosis of malignancy.
The D816V mutation can be detected if tumor represents at least 0.3% of cells.
cKIT (D816V) Mutation by PCR
< BACK
< VIEW MARKER
CLOSE X
CLOSE X
• Confirms the diagnosis and follow-up of CML and ALL
• Genoptix BCR/ABL by PCR detects the three main gene breakpoints:
- Major (p210) – most common breakpoint in CML
- Minor (p190) - in rare cases can be seen in CML with increased monocytes, resembling CMML, and in the majority of ALL
- Micro (p230) – rare, associated with prominent neutrophilic maturation (mimicking CNL) and/or conspicuous thrombocytosis
• Can be identified by three technologies:
- Karyotype – identify presence of t(9;22)
- FISH – may identify presence of BCR/ABL translocation not identified by routine cytogenetics due to cryptic translocations
- PCR - detects and quantifies BCR/ABL transcripts, establishes a baseline at diagnosis and provides a means for follow-up and disease monitoring including assessment of molecular response to TKIs
• Genoptix serial reporting of BCR/ABL by PCR defines the target three-log reduction, and graphs as many as five serial results on one report to help track disease
BCR/ABL
< BACK
VIEW REPORT >
Reference: NCCN Practice Guidelines in Oncology: Chronic Myelogenous Leukemia, Version 2.2011.
CLOSE X
CLINICAL DATA:
46-year-old male with chronic myelogenous leukemia, on a TKI. Previous CHART report on peripheral blood shows trilineage hematopoiesis and molecular evidence of minimal residual CML (collected 08/06/09). Prior COMPASS report on bone marrow reveals CML in chronic phase collected 03/05/09). Peripheral blood submitted for evaluation. Accompanying CBC report, dated 9/16/10, indicates WBC 3.7 K/uL, RBC 3.69 M/uL, Hgb 12.2 g/dL, HCT 37.0%, MCV 100 fL, MCH 33.1 pg, MCHC 33.0 g/dL, RDW 11.1%, platelets 265 K/uL with a differential count of neutrophils 57.9%, lymphocytes 36.6%, monocytes 1.9%, eosinophils 3.0%, basophils 0.6%.
Results:
POSITIVE. The BCR/ABL transcript is detected at 0.055% BCR/ABL as compared to ABL.
46-year-old male with chronic myelogenous leukemia, on a TKI. Previous CHART report on peripheral blood shows trilineage hematopoiesis and molecular evidence of minimal residual CML (collected 08/06/09). Prior COMPASS report on bone marrow reveals CML in chronic phase collected 03/05/09). Peripheral blood submitted for evaluation. Accompanying CBC report, dated 9/16/10, indicates WBC 3.7 K/uL, RBC 3.69 M/uL, Hgb 12.2 g/dL, HCT 37.0%, MCV 100 fL, MCH 33.1 pg, MCHC 33.0 g/dL, RDW 11.1%, platelets 265 K/uL with a differential count of neutrophils 57.9%, lymphocytes 36.6%, monocytes 1.9%, eosinophils 3.0%, basophils 0.6%.
Results:
POSITIVE. The BCR/ABL transcript is detected at 0.055% BCR/ABL as compared to ABL.
GenoTRACE® Report
BCR/ABL t(9;22) Detection by Quantitative RT-PCR
BCR/ABL t(9;22) Detection by Quantitative RT-PCR
< BACK
< VIEW MARKER
Notes:
Quantitative PCR detects the presence of the major/p210 breakpoint (e13a2) BCR/ABL transcript. BCR/ABL is required for the diagnosis of chronic myelogenous leukemia (CML) and is present in ~20% of acute lymphoblastic leukemia (ALL) cases. The therapeutic significance of any one level of BCR/ABL is not clear. However, the trend over time may be useful in management. Studies have shown that achievement of a 3 log reduction in BCR/ABL RNA from baseline median (0.079% at Genoptix) within 18 month of treatment with first generation tyrosine kinase inhibitor (TKI) treatment is associated with a minimal risk of progression at 84 months from the start of treatment. Variable BCR-ABL breakpoints are associated with certain phenotypic features of CML and may confer different prognostic significance. CML patients in chronic phase and carrying predominantly minor BCR/ABL transcript tend to have a more “monocytic” CML, those with a micro breakpoint tend to have a more “neutrophilic” CML.
Quantitative PCR detects the presence of the major/p210 breakpoint (e13a2) BCR/ABL transcript. BCR/ABL is required for the diagnosis of chronic myelogenous leukemia (CML) and is present in ~20% of acute lymphoblastic leukemia (ALL) cases. The therapeutic significance of any one level of BCR/ABL is not clear. However, the trend over time may be useful in management. Studies have shown that achievement of a 3 log reduction in BCR/ABL RNA from baseline median (0.079% at Genoptix) within 18 month of treatment with first generation tyrosine kinase inhibitor (TKI) treatment is associated with a minimal risk of progression at 84 months from the start of treatment. Variable BCR-ABL breakpoints are associated with certain phenotypic features of CML and may confer different prognostic significance. CML patients in chronic phase and carrying predominantly minor BCR/ABL transcript tend to have a more “monocytic” CML, those with a micro breakpoint tend to have a more “neutrophilic” CML.
CLOSE X
CLOSE X
• MPL W515L/K is located on chromosome 1, exon 10
• Present in a subset of patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF)
• MPL W515L/K mutation affects the thrombopoietin receptor (TPO) (see image on page 2)
• TPO is the primary regulator of megakaryocyte maturation and supports production of functional platelets
• Genoptix PCR assay can detect mutant DNA when present at levels as low as 5%
MPL
< BACK
VIEW REPORT >
CLOSE X
MPL W515 L/K Mutation Analysis
< BACK
< VIEW MARKER
CLINICAL DATA:
79-year-old female with anemia, leukocytosis, and a history of thrombocytopenia. Treatment includes Aranesp. Peripheral blood submitted for evaluation. Accompanying CBC report, dated 9/17/08, indicates WBC 12.2 K/uL, RBC 3.40 M/uL, Hgb 9.6 g/dL, HCT 30.5%, MCV 90 fL, MCH 28.2 pg, MCHC 31.5 g/dL, RDW 22.3%, platelets 326 K/uL with a differential count of neutrophils 41%, bands 17%, metamyelocytes 5%, myelocytes 12%, promyelocytes 5%, blasts 2%, lymphocytes 15%, monocytes 3%, eosinophils 0%, basophils 0%.
Results:
Results are POSITIVE for the MPL W515 mutation by Invader Plus®. The test detected the K allele of MPL indicating a TG to AA mutation in codon 515 causing an amino acid change from tryptophan to lysine.
Notes:
Results of MPL W515 testing by Invader Plus® are POSITIVE, indicating a W515 mutation is present in the MPL gene. This mutation, which results in the constitutive cytokine-independent activation of the JAK-STAT pathway, is present in a subset of patients with chronic idiopathic myelofibrosis, essential thrombocythemia, and acute myeloid leukemia with antecedent myeloproliferative disorder(1). MPL codes for the thrombopoietin (TPO) receptor. TPO is the primary regulator of megakaryocyte maturation and supports production of functional platelets (2). Nucleic acid studies do not constitute a definitive diagnostic test for all chronic myeloproliferative neoplasms but, togetherwith other clinicopathologic findings, can support their diagnosis. Specifically, as mixed myeloproliferative syndromes are not uncommon, it is recommended that a careful correlation with bone marrow histology be performed.
Method:
MPL W515 mutation analysis includes isolation of DNA, target gene amplification via the polymerase chain reaction (PCR), and a Cleavase-based allele-specific signal amplification to determine the presence of the W515L (TGG>TTG) and W515K (TGG>AAG) genotypes. As assessed by positive DNA dilution experiments, this assay can detect mutant DNA whenpresent at levels as low as 5%.
Technical Note:
A majority of patients with chronic myeloproliferative neoplasms will be negative for the MPL W515 mutation. Since the mutation typically is a somatic mutation, peripheral blood specimens with predominantly normal lymphocytes, or other cells not carrying the mutation, may have mutation levels below the limit of detection for this assay. Genetic testing errors may result from trace contamination of PCR reactions and from rare genetic variants that interfere with analysis.
79-year-old female with anemia, leukocytosis, and a history of thrombocytopenia. Treatment includes Aranesp. Peripheral blood submitted for evaluation. Accompanying CBC report, dated 9/17/08, indicates WBC 12.2 K/uL, RBC 3.40 M/uL, Hgb 9.6 g/dL, HCT 30.5%, MCV 90 fL, MCH 28.2 pg, MCHC 31.5 g/dL, RDW 22.3%, platelets 326 K/uL with a differential count of neutrophils 41%, bands 17%, metamyelocytes 5%, myelocytes 12%, promyelocytes 5%, blasts 2%, lymphocytes 15%, monocytes 3%, eosinophils 0%, basophils 0%.
Results:
Results are POSITIVE for the MPL W515 mutation by Invader Plus®. The test detected the K allele of MPL indicating a TG to AA mutation in codon 515 causing an amino acid change from tryptophan to lysine.
Notes:
Results of MPL W515 testing by Invader Plus® are POSITIVE, indicating a W515 mutation is present in the MPL gene. This mutation, which results in the constitutive cytokine-independent activation of the JAK-STAT pathway, is present in a subset of patients with chronic idiopathic myelofibrosis, essential thrombocythemia, and acute myeloid leukemia with antecedent myeloproliferative disorder(1). MPL codes for the thrombopoietin (TPO) receptor. TPO is the primary regulator of megakaryocyte maturation and supports production of functional platelets (2). Nucleic acid studies do not constitute a definitive diagnostic test for all chronic myeloproliferative neoplasms but, togetherwith other clinicopathologic findings, can support their diagnosis. Specifically, as mixed myeloproliferative syndromes are not uncommon, it is recommended that a careful correlation with bone marrow histology be performed.
Method:
MPL W515 mutation analysis includes isolation of DNA, target gene amplification via the polymerase chain reaction (PCR), and a Cleavase-based allele-specific signal amplification to determine the presence of the W515L (TGG>TTG) and W515K (TGG>AAG) genotypes. As assessed by positive DNA dilution experiments, this assay can detect mutant DNA whenpresent at levels as low as 5%.
Technical Note:
A majority of patients with chronic myeloproliferative neoplasms will be negative for the MPL W515 mutation. Since the mutation typically is a somatic mutation, peripheral blood specimens with predominantly normal lymphocytes, or other cells not carrying the mutation, may have mutation levels below the limit of detection for this assay. Genetic testing errors may result from trace contamination of PCR reactions and from rare genetic variants that interfere with analysis.
References:
1. Pardanani AD, Levine RL, Lasho T, Pikman Y, Mesa RA, Wadleigh M, Steensma DP, Elliot MA, Wolanskyj AP, Hogan WJ, McClure RF, Litzow MR, Gilliland DG, and Tefferi A. MPL515 mutations in myeloproliferative and other myeloid disorders: a study of 1182 patients. Blood. 2006 Nov 15;108:3472-3476.
2. Duensing S, Duensing A, Meran JG, Kreft A, Busche G, Ganser A, Georgii A. Molecular detection of c-mpl thrombopoietin receptor gene expression in chronic myeloproliferative disorders. Mol Pathol. 1999 Jun;52(3):146-50.
1. Pardanani AD, Levine RL, Lasho T, Pikman Y, Mesa RA, Wadleigh M, Steensma DP, Elliot MA, Wolanskyj AP, Hogan WJ, McClure RF, Litzow MR, Gilliland DG, and Tefferi A. MPL515 mutations in myeloproliferative and other myeloid disorders: a study of 1182 patients. Blood. 2006 Nov 15;108:3472-3476.
2. Duensing S, Duensing A, Meran JG, Kreft A, Busche G, Ganser A, Georgii A. Molecular detection of c-mpl thrombopoietin receptor gene expression in chronic myeloproliferative disorders. Mol Pathol. 1999 Jun;52(3):146-50.
CLOSE X
CLOSE X
• Del(5q) is a sole cytogenetic abnormality
• Predominantly in female patients with a 7:3 ratio
• Favorable prognosis with a 12% to 16% transformation rate to AML and median survival of >5 years
• Clinical presentation is macrocytic anemia, mild leukopenia, normal or increased platelet count
• Causes treatment-resistant anemia and MDS
5q-
< BACK
References:
Boultwood J, et al. Blood. 1994;84:3253-3260.
Mathew P, et al. Blood. 1993:81:1040-1045.
Boultwood J, et al. Blood. 1994;84:3253-3260.
Mathew P, et al. Blood. 1993:81:1040-1045.
CLOSE X
CLOSE X
• Described as "the guardian of the genome” for its role in conserving stability by preventing genome mutation1
• A tumor suppressor gene located on the short arm of chromosome 172
• Single P53 deletion should be considered as a high-risk aberration for future risk-adapted treatment strategies in AML2
• The molecular risk factors FLT3-ITD and NPM1 mutation have shown an inverse correlation to the P53 deletion2
• Somatic mutations of P53 can be found in half of all cancers1
17p (P53)
< BACK
References:
1. Genetic Home Reference: TP53. National Institute of Health. http://ghr.nlm.nih.gov/gene/TP53. Accessed 11-16-11
2. Seifert H, Mohr B, Thiede C, Et. Al. The prognostic impact of 17p (p53) deletion in 2272 adults with acute myeloid leukemia. Leukemia. 2009:23, 656-663.
1. Genetic Home Reference: TP53. National Institute of Health. http://ghr.nlm.nih.gov/gene/TP53. Accessed 11-16-11
2. Seifert H, Mohr B, Thiede C, Et. Al. The prognostic impact of 17p (p53) deletion in 2272 adults with acute myeloid leukemia. Leukemia. 2009:23, 656-663.
CLOSE X