CAPFITOGEN tools. Facilitated spatial and ecogeographical germplasm analysis ...Mauricio Parra Quijano
Presentation from the conference ENHANCED GENEPOOL UTILIZATION ‒ Capturing wild relative and landrace diversity for crop improvement, June 2014, Cambridge, UK
CAPFITOGEN tools. Facilitated spatial and ecogeographical germplasm analysis ...Mauricio Parra Quijano
Presentation from the conference ENHANCED GENEPOOL UTILIZATION ‒ Capturing wild relative and landrace diversity for crop improvement, June 2014, Cambridge, UK
CAPFITOGEN Programme for the Strengthening of Capabilities in National Plant Genetic Resources Programmes, International Treaty on Plant Genetic Resources for Food and Agriculture - FAO
"Bacterial Pathogen Genomics at NCBI" presentation at the Standards for Pathogen Identification via NGS (SPIN) workshop hosted by the National Institute for Standards and Technology October 2014 by Dr. Bill Klimke.
Ion Torrent™ semiconductor sequencing, combined with Ion AmpliSeq™ technology, provides simultaneous identification of copy number variants (CNVs), single nucleotide variants (SNVs), and small insertions and deletions (indels) from a research sample by means of a single integrated workflow. 100% of assayed CNV regions (n=34) were detected using a reference set of 31 samples with known chromosomal aberrations. Low-pass whole-genome sequencing data, with approximately 0.01x read coverage, allowed the rapid ≤10 hour analysis of aneuploidies from research samples with extremely low initial input DNA amounts—even from a single cell. Using a control set of 10 samples with known chromosomal aberrations, 100% of the copy number changes were found, ranging from gains or losses of whole chromosomes to subchromosomal alterations tens of megabases (Mb) in size. The Ion PGM™ System minimizes the high cost and complexity of next-generation sequencing and, with Ion Reporter™ Software, facilitates user-defined CNV and aneuploidy detection, with three sensitivity options so that copy number analysis workflows can be tuned to achieve desired levels of sensitivity and specificity.
CAPFITOGEN Programme for the Strengthening of Capabilities in National Plant Genetic Resources Programmes, International Treaty on Plant Genetic Resources for Food and Agriculture - FAO
Automated Diagnosis of Malaria in Tropical Areas Using 40X Microscopic Images...CSCJournals
This paper proposes a new algorithm to measure parasitemia stemming from Plasmodium
falciparum by using optical images of capillary blood smears from Sub-Saharan Africa. The
approach is an improvement of the previous noteworthy method by developing a two-tone
adaptive median filter and Sauvola segmentation. The analysis was performed on the database
of 100X and 40X microscopic images originating from real time collection of patients’ blood of
some Cameroon’s laboratories. The obtained results were very satisfactory with a detection of
infected cells on 40X images of a sensitivity at 81.58%, a specificity at 97.11% and an accuracy
at 96.71%. Compared to the previous works, the values portray a real improvement in most
performance’s criteria for 100X images and 40X images as well. There is a significant step of
automated Malaria diagnosis in tropical areas and in the performance assessed. The results of
the method gave more strength for the two magnification ranges. The time spent in analysis is
obviously reduced with 40X magnification and the inaccessibility of the information by the visual
laboratory measurement is henceforth genuinely assaulted. The low cost of the system opens
therefore the possibility of a cost effective method of diagnosing malaria in low and middleincome
countries
During the recent East and Southern Africa review and planning meeting, team members reviewed the Value chain assessment framework – all crops and programs.
Detection of somatic mutations at 0.5% frequency from cfDNA and CTC DNA using...Thermo Fisher Scientific
Availability of effective blood screening for tracking of recurrence and
resistance of tumors may improve outcomes in the future. Research
studies suggest that virtually all tumors carry somatic DNA mutations, and
these may serve as biomarkers that may be tracked from blood. The
two well-characterized sources of tumor DNA in blood are circulating
tumor cells (CTC) and cell-free tumor DNA (ctDNA). The abundance of
CTC and/or ctDNA in blood may be very low at critical stages such as
early recurrence or development of resistance. Hence there is great
interest in being able to detect biomarkers at very low frequency from
blood, and in characterizing the relationship between somatic mutations
present in the tumor and those in CTC or ctDNA.
We present a research use only analysis workflow for peripheral
monitoring that enables detection of low frequency variants in blood. We
developed an analysis algorithm, using statistical modeling of next
generation sequencing reads, and optimizing parameters and filters to
enable sensitive and specific detection of somatic mutations at 0.5% allele
ratio. We demonstrate the analysis on a blood sample split into 3 subsamples
comprising normal blood, CTC enriched, and cell-free DNA
(cfDNA) samples.
We used lysis to isolate white blood cells (germline), centrifugation to
extract plasma DNA (cfDNA), while CTC cells were isolated using
Cynvenio LiquidBiopsy™ platform a fully automated antibody-based
solution. We barcoded 3 sub-samples and run them on a single Ion 318™
sequencing chip using Ion AmpliSeq™ Cancer Hotspot Panel (CHPv2),
that enables very deep (~10,000x coverage) and accurate
sequencing. This panel allows interrogation of ~2800 relevant
biomarkers from COSMIC and FDA actionable databases, and denovo
variant detection at ~20,000 genomic positions. Mutations were
annotated using the Oncomine® database in Ion Reporter™ software. The
research assay requires a small amount of input DNA (~10ng), and has a
fast turn around time from extracted DNA to variants of less than 24 hr.
Bioinformatics tools for the diagnostic laboratory - T.Seemann - Antimicrobi...Torsten Seemann
"Bioinformatics tools for the diagnostic laboratory" presented at the Australian Society for Antimicrobials 2016 annual conference in Melbourne Australia. Slides are aimed at a biological / pathology / clinican audience. Some material has been re-imagined from Nick Loman's ECCMID 2015 talk.
Principle, Procedure and applications of Digital PCR.pptxVikramadityaupmanyu
Digital PCR (dPCR) is the new generation PCR that enables absolute quantification of target gene by separating reac¬tion mixture in several compartments. In this system, copies of target nucleic acid are distributed randomly from 0, 1 or many in the several small volume compartments. Amplification is occurred in the compartment and resulting absorbance is measured. Integrated fluidic circuits, chip based microchambers, and water in oil droplets are the methods are used for separation of reaction mixture in to several compartments. BioMark HD system (Fluidigm, USA) and QuantStudio3D system (Thermofisher Scientific, USA) uses integrated fluidic circuits harbor¬ing 10000 to 40000 microchambers and integrated chip containing 20000 microchambers, respectively. In droplet digital PCR System (Biorad, USA), reaction mixture is separated into 20,000 water in droplets. After cycles of reaction using any of the above technologies, fluorescence is detected with an imaging sys¬tem in the each compartment and copy numbers of the target is calculated with imaging software.
In droplets digital PCR, after amplification, droplets containing target gene are detected by fluorescence and scored as positive, and droplets without fluorescence are scored as negative. Poisson statistical analysis of the numbers of positive and negative droplets yields absolute quantitation of the target sequence. Digital PCR is the preferred technique for absolute quantification of target gene without need of standard curve and higher sensitivity and produces highly reproducible results, and also less susceptible to inhibitors than conventional RT-qPCR
The OncoScan(TM) platform for analysis of copy number and somatic mutations i...Lawrence Greenfield
The OncoScan microarray offers high-quality copy number, genotype, and somatic mutation data with whole-genome coverage and high resolution in cancer genes for use with challenging FFPE samples.
"Bacterial Pathogen Genomics at NCBI" presentation at the Standards for Pathogen Identification via NGS (SPIN) workshop hosted by National Institute for Standards and Technology October 2014 by Dr. Bill Klimke.
CAPFITOGEN Programme for the Strengthening of Capabilities in National Plant Genetic Resources Programmes, International Treaty on Plant Genetic Resources for Food and Agriculture - FAO
Aplicaciones y herramientas de ecogeografía para la colecta, conservación y u...Mauricio Parra Quijano
Presentación ofrecida en el 2do. Congreso Nacional de Recursos Fitogenéticos y 1er. Congreso Internacional Conservación y Aprovechamiento sustentable de la Agrobiodiversidad. Universidad de Chapingo, Texcoco, México, 25 al 27 de Noviembre de 2015.
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Ion Torrent™ semiconductor sequencing, combined with Ion AmpliSeq™ technology, provides simultaneous identification of copy number variants (CNVs), single nucleotide variants (SNVs), and small insertions and deletions (indels) from a research sample by means of a single integrated workflow. 100% of assayed CNV regions (n=34) were detected using a reference set of 31 samples with known chromosomal aberrations. Low-pass whole-genome sequencing data, with approximately 0.01x read coverage, allowed the rapid ≤10 hour analysis of aneuploidies from research samples with extremely low initial input DNA amounts—even from a single cell. Using a control set of 10 samples with known chromosomal aberrations, 100% of the copy number changes were found, ranging from gains or losses of whole chromosomes to subchromosomal alterations tens of megabases (Mb) in size. The Ion PGM™ System minimizes the high cost and complexity of next-generation sequencing and, with Ion Reporter™ Software, facilitates user-defined CNV and aneuploidy detection, with three sensitivity options so that copy number analysis workflows can be tuned to achieve desired levels of sensitivity and specificity.
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This paper proposes a new algorithm to measure parasitemia stemming from Plasmodium
falciparum by using optical images of capillary blood smears from Sub-Saharan Africa. The
approach is an improvement of the previous noteworthy method by developing a two-tone
adaptive median filter and Sauvola segmentation. The analysis was performed on the database
of 100X and 40X microscopic images originating from real time collection of patients’ blood of
some Cameroon’s laboratories. The obtained results were very satisfactory with a detection of
infected cells on 40X images of a sensitivity at 81.58%, a specificity at 97.11% and an accuracy
at 96.71%. Compared to the previous works, the values portray a real improvement in most
performance’s criteria for 100X images and 40X images as well. There is a significant step of
automated Malaria diagnosis in tropical areas and in the performance assessed. The results of
the method gave more strength for the two magnification ranges. The time spent in analysis is
obviously reduced with 40X magnification and the inaccessibility of the information by the visual
laboratory measurement is henceforth genuinely assaulted. The low cost of the system opens
therefore the possibility of a cost effective method of diagnosing malaria in low and middleincome
countries
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Availability of effective blood screening for tracking of recurrence and
resistance of tumors may improve outcomes in the future. Research
studies suggest that virtually all tumors carry somatic DNA mutations, and
these may serve as biomarkers that may be tracked from blood. The
two well-characterized sources of tumor DNA in blood are circulating
tumor cells (CTC) and cell-free tumor DNA (ctDNA). The abundance of
CTC and/or ctDNA in blood may be very low at critical stages such as
early recurrence or development of resistance. Hence there is great
interest in being able to detect biomarkers at very low frequency from
blood, and in characterizing the relationship between somatic mutations
present in the tumor and those in CTC or ctDNA.
We present a research use only analysis workflow for peripheral
monitoring that enables detection of low frequency variants in blood. We
developed an analysis algorithm, using statistical modeling of next
generation sequencing reads, and optimizing parameters and filters to
enable sensitive and specific detection of somatic mutations at 0.5% allele
ratio. We demonstrate the analysis on a blood sample split into 3 subsamples
comprising normal blood, CTC enriched, and cell-free DNA
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extract plasma DNA (cfDNA), while CTC cells were isolated using
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solution. We barcoded 3 sub-samples and run them on a single Ion 318™
sequencing chip using Ion AmpliSeq™ Cancer Hotspot Panel (CHPv2),
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sequencing. This panel allows interrogation of ~2800 relevant
biomarkers from COSMIC and FDA actionable databases, and denovo
variant detection at ~20,000 genomic positions. Mutations were
annotated using the Oncomine® database in Ion Reporter™ software. The
research assay requires a small amount of input DNA (~10ng), and has a
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Presentation3 - Representa & DIVmapas tools
1. Mauricio Parra Quijano
FAO consultant
International Treaty on Plant Genetic Resources
for Nutrition and Agriculture
CAPFITOGEN Program Coordinator
Tools
13. DIVmapas
Selection of cells that include collecting sites (centroids are points in
yellow )
14. DIVmapas
Detection of neighbouring cells which centroids (green points) falls at X km
(radius ) around of the presence cell centroid (yellow points)
15. DIVmapas
Generation of areas of influence from each centroid
Detection of accessions collected within each area of influence