Development of Laser-Mediated Nanodroplet Real-Time PCR on Circulating Tumor Cells (CTC) by Microfilter Platform

2015 ◽  
Author(s):  
Gregory W. Faris
Keyword(s):  
Real Time ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Real Time Pcr

scholarly journals Molecular characterization of circulating tumor cells in large quantities of contaminating leukocytes by a multiplex real-time PCR

2008 ◽  
Vol 118 (3) ◽  
pp. 455-468 ◽  
Author(s):  
Anieta M. Sieuwerts ◽  
Jaco Kraan ◽  
Joan Bolt-de Vries ◽  
Petra van der Spoel ◽  
Bianca Mostert ◽  
...  
Keyword(s):  
Real Time ◽  
Tumor Cells ◽  
Molecular Characterization ◽  
Circulating Tumor Cells ◽  
Real Time Pcr

scholarly journals FusionCatcher - a tool for finding somatic fusion genes in paired-end RNA-sequencing data

2014 ◽  
Author(s):  
Daniel Nicorici ◽  
Mihaela Satalan ◽  
Henrik Edgren ◽  
Sara Kangaspeska ◽  
Astrid Murumagi ◽  
...  
Keyword(s):  
Real Time ◽  
Tumor Cells ◽  
Rna Sequencing ◽  
Real Time Pcr ◽  
Software Tool ◽  
Fusion Genes ◽  
Sequencing Data ◽  
Detection Rates ◽  
Somatic Fusion

FusionCatcher is a software tool for finding somatic fusion genes in paired-end RNA-sequencing data from human or other vertebrates. FusionCatcher achieves competitive detection rates and real-time PCR validation rates in RNA-sequencing data from tumor cells. FusionCatcher is available at http://code.google.com/p/fusioncatcher

1327 Real-time risk evaluation of metastasis using circulating tumor cells

2009 ◽  
Vol 7 (2) ◽  
pp. 147 ◽  
Author(s):  
A. de Albuquerque ◽  
S. Kaul ◽  
N. Fersis ◽  
D. Ernst ◽  
A. Teubner ◽  
...  
Keyword(s):  
Real Time ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Risk Evaluation

BRAF V600E Mutation Appears Specific for Hairy Cell Leukemia Among Low and Intermediate Grade B-Cell Lymphomas: Utility of a Real Time PCR Based Approach for Detection

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2635-2635
Author(s):  
Mark Ewalt ◽  
Subhadra Nandula ◽  
Adrienne A. Phillips ◽  
Vundavalli Murty ◽  
Mahesh Mansukhani ◽  
...  
Keyword(s):  
Real Time ◽  
Tumor Cells ◽  
Real Time Pcr ◽  
Braf V600e Mutation ◽  
Braf V600e ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Pcr Assay ◽  
Neoplastic Cells ◽  
Intermediate Grade

Abstract Abstract 2635 BACKGROUND: Hairy cell leukemia (HCL) is a rare type of B-cell non Hodgkin lymphoma (B-NHL), which is characterized by neoplastic cells exhibiting slender cytoplasmic projections and an activated phenotype. Unlike other B-NHL, HCL lacks characteristic recurrent chromosome abnormalities and its etiology remains elusive. Recently, the BRAF V600E mutation was described to occur at a high frequency in HCL in contrast to other B-NHL, suggesting that the BRAF V600E mutation could represent a disease defining lesion and a possible therapeutic target. In this study, we determined the prevalence of the BRAF V600E mutation in a large series of low and intermediate grade B-NHL and assessed the relationship of this mutation, if any, with microsatellite instability (MSI). METHODS: DNA was extracted from various low and intermediate grade B-NHL (n=100), as defined by the 2008 World Health Organization Classification, which comprised HCL (n=10 from 6 patients), chronic lymphocytic leukemia/small lymphocytic lymphoma (n=22), mantle cell lymphoma (n=19), marginal zone lymphoma (n=22), follicular lymphoma (n=20) and lymphoplasmacytic lymphoma (n=7). Percentage of neoplastic cells was assessed by flow cytometry (n=99) or immunohistochemistry (n=1). Presence of BRAF V600E mutation was determined using a real time PCR assay using allele-specific hydrolysis (“Taqman”) probes. The detection limit of the assay was determined by diluting DNA extracted from a fresh HCL sample with normal high molecular weight DNA extracted by the same method. MSI analysis was performed using a multiplex reaction for 5 quasi monomorphic mononucleotide repeat markers (BAT-25, BAT-26, MONO-27, NR-21 and NR-24) and two highly polymorphic penta nucleotide markers (Penta C and Penta D) as sample identifiers. In the absence of normal DNA for comparison, a sample was considered MSI-H if greater than two markers showed an altered pattern, indeterminate if only two showed an alteration and MSS if no marker showed an altered allele. RESULTS: The ubiquitous occurrence and high specificity of the BRAF V600E mutation for HCL was confirmed in our series of B-NHL. The HCL samples consisted of 10 specimens from 6 patients and were the initial diagnostic specimen in 2 patients. The percentage of neoplastic cells in these samples ranged from 1% to 78.4% (median 18.2%). The BRAF V600E mutation was detected in 7 of 10 (70%) samples of HCL from 5 of 6 (83%) patients. The proportion of hairy cells was ≤5% in samples where the mutation was not detected. All 90 of the other low and intermediate grade B-NHL (>50% tumor burden) were negative for the V600E mutation. Compared to the reported sensitivity of approximately 30% tumor cells for detecting the BRAF V600E mutation by Sanger sequencing, our real time PCR assay allowed detection of the mutation in samples containing ≥9.8% tumor cells. Analysis of the HCL cases for MSI revealed that all cases had a microsatellite stable (MSS) phenotype. CONCLUSIONS: The BRAF V600E mutation appears specific for HCL among low and intermediate grade B-NHL and is not associated with microsatellite instability. These characteristics thus warrant inclusion in disease definition. Further refinements of a realtime PCR based approach for detecting the BRAF V600E mutation might be of utility in diagnosis and disease monitoring, as the neoplastic cellular yield is often limited in HCL due to underlying myelofibrosis. Disclosures: No relevant conflicts of interest to declare.

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