scholarly journals Concentration of Plasmodium falciparum gametocytes in whole blood samples by magnetic cell sorting enhances parasite infection rates in mosquito feeding assays

2017 ◽  
Vol 16 (1) ◽  
Author(s):  
Isaie J. Reuling ◽  
Will J. R. Stone ◽  
Marga van de Vegte-Bolmer ◽  
Geert-Jan van Gemert ◽  
Rianne Siebelink-Stoter ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Cell Sorting ◽  
Whole Blood ◽  
Parasite Infection ◽  
Infection Rates ◽  
Magnetic Cell Sorting ◽  
Blood Samples ◽  
Whole Blood Samples

scholarly journals Using CF11 cellulose columns to inexpensively and effectively remove human DNA from Plasmodium falciparum-infected whole blood samples

2012 ◽  
Vol 11 (1) ◽  
pp. 41 ◽  
Author(s):  
Meera Venkatesan ◽  
Chanaki Amaratunga ◽  
Susana Campino ◽  
Sarah Auburn ◽  
Oliver Koch ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Whole Blood ◽  
Blood Samples ◽  
Human Dna ◽  
Whole Blood Samples ◽  
Cf11 Cellulose

scholarly journals 5WBF: A low-cost and straightforward whole blood filtration method suitable for whole-genome sequencing of Plasmodium falciparum clinical isolates

2021 ◽  
Author(s):  
Romain Coppée ◽  
Atikatou Mama ◽  
Véronique Sarrasin ◽  
Claire Kamaliddin ◽  
Lucie Adoux ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Whole Blood ◽  
Nuclear Genome ◽  
Whole Genome ◽  
Ring Stage ◽  
Filtration Method ◽  
Blood Samples ◽  
Organellar Genomes ◽  
Whole Blood Samples ◽  
Template Dna

Background: Whole-genome sequencing (WGS) is becoming increasingly helpful to assist malaria control programs. A major drawback of this approach is the large amount of human DNA compared to parasite DNA extracted from unprocessed whole blood. As red blood cells (RBCs) have a diameter of about 7-8 μm and exhibit some deformability, we hypothesized that cheap and commercially available 5 μm filters might retain leukocytes but much less of Plasmodium falciparum-infected RBCs. This study aimed to test the hypothesis that such a filtration method, named 5WBF (for 5 μm Whole Blood Filtration), may provide highly enriched parasite material suitable for P. falciparum WGS. Methods: Whole blood was collected from five patients experiencing a P. falciparum malaria episode (ring-stage parasitemia range: 0.04-5.5%) and from mock samples obtained by mixing synchronized, ring-stage cultured P. falciparum 3D7 parasites with uninfected human whole blood (final parasitemia range: 0.02-1.1%). These whole blood samples (50 to 400 μL) were diluted in RPMI 1640 medium or PBS 1X buffer and filtered with syringes connected to a 5 μm commercial filter. DNA was extracted from filtered and unfiltered counterpart blood samples using a commercial kit. The 5WBF method was evaluated on the ratios of parasite:human DNA assessed by qPCR and by sequencing depth and percentages of coverage from WGS data (Illumina NextSeq 500). As a comparison, we also applied to the same unprocessed whole blood samples the selective whole-genome amplification (sWGA) method which does not rely on blood filtration. Results: After applying 5WBF, qPCR indicated an average of 2-fold loss in the amount of parasite template DNA (Pf ARN18S gene) and from 4,096- to 65,536-fold loss of human template DNA (human β actin gene). WGS analyses revealed that > 95% of the nuclear genome and the entire whole organellar genomes were covered at ≥ 10x depth for all samples tested. In sWGA counterparts, none of the organellar genomes were covered, and from 47.7 to 82.1% of the nuclear genome was covered at ≥ 10x depth depending on parasitemia. Sequence reads were homogeneously distributed across gene sequences for 5WBF-treated samples (n = 5,460 genes; mean coverage: 91x; median coverage: 93x; 5th percentile: 70x; 95th percentile: 103x), allowing the identification of gene copy number variations such as for gch1. This later analysis was not possible for sWGA-treated samples, as we observed a much more heterogeneous distribution of reads among gene sequences (mean coverage: 80x; median coverage: 51x; 5th percentile: 7x; 95th percentile: 245x. Conclusions: The novel 5WBF leucodepletion method is simple to implement and based on commercially available, standardized, 5 μm filters which cost from 1.0 to 1.7€ per unit, depending on suppliers. 5WBF permits extensive genome-wide analysis of P. falciparum DNA from minute amounts of whole blood even with parasitemias as low as 0.02%.

Steadiness/stability of antiepileptic drugs in serum, plasma and whole-blood samples under various storage methods

2010 ◽  
Vol 41 (02) ◽  
Author(s):  
N Shazi ◽  
A Böss ◽  
HJ Merkel ◽  
F Scharbert ◽  
D Hannak ◽  
...  
Keyword(s):  
Antiepileptic Drugs ◽  
Whole Blood ◽  
Blood Samples ◽  
Storage Methods ◽  
Whole Blood Samples

scholarly journals Determination of 19 Psychoactive Substances in Premortem and Postmortem Whole Blood Samples Using Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Separations ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 78
Author(s):  
Sevasti Karampela ◽  
Jessica Smith ◽  
Irene Panderi
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Formic Acid ◽  
Whole Blood ◽  
High Performance ◽  
Tandem Mass ◽  
Psychoactive Substances ◽  
Blood Samples ◽  
Whole Blood Samples

An ever-increasing need exists within the forensic laboratories to develop analytical processes for the qualitative and quantitative determination of a broad spectrum of new psychoactive substances. Phenylethylamine derivatives are among the major classes of psychoactive substances available on the global market and include both amphetamine analogues and synthetic cathinones. In this work, an ultra-high-performance liquid chromatography-positive ion electrospray ionization tandem mass spectrometric method (UHPLC-ESI-MS/MS) has been developed and fully validated for the determination of 19 psychoactive substances, including nine amphetamine-type stimulants and 10 synthetic cathinone derivatives, in premortem and postmortem whole blood. The assay was based on the use of 1 mL premortem or postmortem whole blood, following solid phase extraction prior to the analysis. The separation was achieved on a Poroshell 120 EC-C18 analytical column with a gradient mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water in 9 min. The dynamic multiple reaction monitoring used in this work allowed for limit of detection (LOD) and lower limit of quantitation (LOQ) values of 0.5 and 2 ng mL−1, respectively, for all analytes both in premortem and postmortem whole blood samples. A quadratic calibration model was used for the 12 quantitative analytes over the concentration range of 20–2000 ng mL−1, and the method was shown to be precise and accurate both in premortem and postmortem whole blood. The method was applied to the analysis of real cases and proved to be a valuable tool in forensic and clinical toxicology.

scholarly journals Isolation and characterization of circulating pro-vascular progenitor cell subsets from human whole blood samples

2021 ◽  
Vol 2 (1) ◽  
pp. 100311
Author(s):  
Daniella C. Terenzi ◽  
Ehab Bakbak ◽  
Justin Z. Trac ◽  
Mohammad Al-Omran ◽  
Adrian Quan ◽  
...  
Keyword(s):  
Whole Blood ◽  
Progenitor Cell ◽  
Blood Samples ◽  
Human Whole Blood ◽  
Isolation And Characterization ◽  
Whole Blood Samples
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