Comparative sensitivity analyses of quantitative polymerase chain reaction and flow cytometry in detecting cellular microchimerism in murine tissues

2014 ◽  
Vol 406 ◽  
pp. 74-82 ◽  
Author(s):  
Kristin Thiele ◽  
Carsten Holzmann ◽  
Maria Emilia Solano ◽  
Gunther Zahner ◽  
Petra Clara Arck
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Sensitivity Analyses ◽  
Chain Reaction ◽  
Comparative Sensitivity ◽  
Polymerase Chain

scholarly journals CURRENT STRATEGIES FOR THE DETECTION OF MINIMAL RESIDUAL DISEASE IN CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA

2016 ◽  
Vol 8 ◽  
pp. 2016024 ◽  
Author(s):  
Juliana Maria Camargos Rocha ◽  
Sandra Guerra Xavier ◽  
Marcelo Eduardo Lima Souza ◽  
Juliana Godoy Assumpção ◽  
Mitiko Murao ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
High Risk ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Minimal Residual

Acute lymphoblastic leukemia (ALL) is the most common cancer in children. Current treatment strategies for childhood ALL result in long term remission for approximately 90% of patients. However, therapeutic response is worse among those who relapse. Several risk stratification approaches based on clinical and biological aspects have been proposed in order to intensify treatment in patients with high risk of relapse and reduce toxicity on those with greater probability of cure.The detection of residual leukemic cells (minimal residual disease, MRD) is the most important prognostic factor to identify high risk patients, allowing redefinition of chemotherapy. In the last decades, several standardized research protocols evaluated MRD using immunophenotyping by flow cytometry and/or real time quantitative polymerase chain reaction at different time points during treatment. Both methods are highly sensitive (10-3 a 10-5), but expensive, complex, and, because of that, require qualified staff and frequently are restricted to reference centers.The aim of this article was to review technical aspects of immunophenotyping by flow cytometry and real time quantitative polymerase chain reaction to evaluate MRD in ALL. 

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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