scholarly journals Analisis Perbandingan Kualitas Produk Amplikon Gen PMSA2 Antara Spesimen Spot Darah Kering dan Vena

2020 ◽  
Vol 12 (1) ◽  
pp. 10-18
Author(s):  
Arsyam Mawardi ◽  
Hendra K. Maury ◽  
Yustinus Maladan
Keyword(s):  
Genomic Dna ◽  
Venous Blood ◽  
Dried Blood Spot ◽  
Laboratory Research ◽  
Blood Samples ◽  
Storage Method ◽  
Vacuum Tubes ◽  
Blood Specimens ◽  
Blood Spot

This study is aimed to analyze the comparative quality of PMSA2 gene amplicon product stability from two different specimen sources, spot specimens of dried blood and venous blood, as well as selecting the best storage method for specimens of blood samples. This research uses descriptive laboratory research methods. The research began with the process of sample preparation for dried blood spot and venous blood, each using Whatman 903 paper and vacuum tubes containing EDTA, isolating genomic DNA using KIT Zymo Research, amplification of PMSA-2 genes with PCR, detection of PCR products through electrophoresis, measurement of DNA concentration and absorbance, and data analysis. The results of this study are expected to be a source of information about the advantages of two specimen storage methods for clinical blood samples, as well as providing a clear description of the quality of each specimen storage method based on the quality of its amplicon products. The results showed that a total of ten medical samples of dried blood spot and ten venous blood were isolated from the genomic DNA of ten and nine, respectively. PMSA2 gene amplicons detected were seven in venous blood and six in dried blood spot. Venous blood specimens have sensitivity in detecting PMSA genes in samples with the highest value of 554 ng / μL and purity of 2,007 (WB7), and concentration of 550.2 and highest purity of 2,076 (WB10). Venous blood storage techniques using categorized vacuum tubes are effective in the detection of PMSA2 genes and have time efficiency in the process. From these results it was also concluded that the comparison analysis of amplicon products between venous blood specimens was better, more stable and efficient than dry blood spot specimens, thus recommending storage of venous blood specimens using vacuum tubes as the best storage method of blood sample specimens.

PKP-014 Dried blood spot sampling of nilotinib in patients with chronic myeloid leukaemia: a comparison with venous blood samples

2014 ◽  
Vol 21 (Suppl 1) ◽  
pp. A142.1-A142
Author(s):  
AM Schimmel ◽  
CCLM Boons ◽  
A Chahbouni ◽  
AJ Wilhelm ◽  
YM Den Hartog ◽  
...  
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Venous Blood ◽  
Dried Blood Spot ◽  
Blood Samples ◽  
Blood Spot

scholarly journals Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking

2018 ◽  
Vol 2 ◽  
pp. 57 ◽  
Author(s):  
Abhinendra Kumar ◽  
Sharayu Mhatre ◽  
Sheela Godbole ◽  
Prabhat Jha ◽  
Rajesh Dikshit
Keyword(s):  
Genomic Dna ◽  
Large Scale ◽  
Venous Blood ◽  
Magnetic Bead ◽  
Epidemiological Studies ◽  
Dried Blood Spot ◽  
Future Research ◽  
Prolonged Storage ◽  
Whole Genome ◽  
Blood Spot

Background: Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies.  Methods: The purpose of this study was to extract the maximum amount of gDNA from DBS on Whatman 903 protein saver card. gDNA was extracted through column  (Qiagen) & magnetic bead based (Invitrogen) methods. Quantification of extracted gDNA was performed with a spectrophotometer, fluorometer, and integrity analyzed by agarose gel electrophoresis.  Result: Large variation was observed in quantity & purity (260/280 ratio, 1.8-2.9) of the extracted gDNA. The intact gDNA bands on the electrophoresis gel reflect the robustness of DBS for gDNA even after prolonged storage time. The extracted gDNA amount 2.16 – 24 ng/µl is sufficient for its PCR based downstream application, but unfortunately it can’t be used for whole genome sequencing or genotyping from extracted gDNA. Sequencing or genotyping can be achieved by after increasing template copy number through whole genome amplification of extracted gDNA. The obtained results create a base for future research to develop high-throughput research and extraction methods from blood samples. Conclusion: The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies.

scholarly journals Measurement of the alcohol biomarker phosphatidylethanol (PEth) in dried blood spots and venous blood—importance of inhibition of post-sampling formation from ethanol

2021 ◽  
Author(s):  
Olof Beck ◽  
Maria Mellring ◽  
Christian Löwbeer ◽  
Sabina Seferaj ◽  
Anders Helander
Keyword(s):  
Filter Paper ◽  
Whole Blood ◽  
Room Temperature ◽  
Venous Blood ◽  
Dried Blood Spots ◽  
Blood Samples ◽  
Ethanol Formation ◽  
Alcohol Biomarker ◽  
Blood Specimens ◽  
Blood Spot

AbstractPhosphatidylethanol (PEth) is a group of phospholipids formed in cell membranes following alcohol consumption by action of the enzyme phospholipase D (PLD). PEth measurement in whole blood samples is established as a specific alcohol biomarker with clinical and forensic applications. However, in blood specimens containing ethanol, formation of PEth may continue after sampling leading to falsely elevated concentrations. This study evaluated the use of dried blood spot (DBS) and microsampling specimens to avoid post-sampling formation of PEth. Filter paper cards and three commercial devices for volumetric microsampling of finger-pricked blood were assessed, using PEth-negative and PEth-positive whole blood fortified with 2 g/L ethanol. PEth (16:0/18:1) was measured by LC–MS/MS. Post-sampling formation of PEth occurred in wet blood and in the volumetric devices, but not filter paper cards, when stored at room temperature for 48 h. Addition of an inhibitor of PLD, sodium metavanadate (NaVO3), eliminated post-sampling formation during storage and drying. In conclusion, the present study confirmed previous observations that PEth can be formed in blood samples after collection, if the specimen contains ethanol. The results further demonstrated that post-sampling formation of PEth from ethanol also occurred with commercial devices for volumetric dried blood microsampling. In order for a PEth result not to be questioned, it is recommended to use a PLD inhibitor, whether venous blood is collected in a vacutainer tube or finger-pricked blood is obtained using devices for dried blood microsampling. Graphical abstract

scholarly journals Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking

2019 ◽  
Vol 2 ◽  
pp. 57
Author(s):  
Abhinendra Kumar ◽  
Sharayu Mhatre ◽  
Sheela Godbole ◽  
Prabhat Jha ◽  
Rajesh Dikshit
Keyword(s):  
Genomic Dna ◽  
Large Scale ◽  
Venous Blood ◽  
Magnetic Bead ◽  
Epidemiological Studies ◽  
Dried Blood Spot ◽  
Future Research ◽  
Prolonged Storage ◽  
Whole Genome ◽  
Blood Spot

Background: Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies.  Methods: The purpose of this study was to extract the maximum amount of gDNA from DBS on Whatman 903 protein saver card. gDNA was extracted through column  (Qiagen) & magnetic bead based (Invitrogen) methods. Quantification of extracted gDNA was performed with a spectrophotometer, fluorometer, and integrity analyzed by agarose gel electrophoresis.  Result: Large variation was observed in quantity & purity (260/280 ratio, 1.8-2.9) of the extracted gDNA. The intact gDNA bands on the electrophoresis gel reflect the robustness of DBS for gDNA even after prolonged storage time. The extracted gDNA amount 2.16 – 24 ng/µl is sufficient for its PCR based downstream application, but unfortunately it can’t be used for whole genome sequencing or genotyping from extracted gDNA. Sequencing or genotyping can be achieved by after increasing template copy number through whole genome amplification of extracted gDNA. The obtained results create a base for future research to develop high-throughput research and extraction methods from blood samples. Conclusion: The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies.

scholarly journals Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking

2019 ◽  
Vol 2 ◽  
pp. 57 ◽  
Author(s):  
Abhinendra Kumar ◽  
Sharayu Mhatre ◽  
Sheela Godbole ◽  
Prabhat Jha ◽  
Rajesh Dikshit
Keyword(s):  
Genomic Dna ◽  
Large Scale ◽  
Venous Blood ◽  
Magnetic Bead ◽  
Epidemiological Studies ◽  
Dried Blood Spot ◽  
Future Research ◽  
Prolonged Storage ◽  
Whole Genome ◽  
Blood Spot

Background: Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies.  Methods: The purpose of this study was to extract the maximum amount of gDNA from DBS on Whatman 903 protein saver card. gDNA was extracted through column  (Qiagen) & magnetic bead based (Invitrogen) methods. Quantification of extracted gDNA was performed with a spectrophotometer, fluorometer, and integrity analyzed by agarose gel electrophoresis.  Result: Large variation was observed in quantity & purity (260/280 ratio, 1.8-2.9) of the extracted gDNA. The intact gDNA bands on the electrophoresis gel reflect the robustness of DBS for gDNA even after prolonged storage time. The extracted gDNA amount 2.16 – 24 ng/µl is sufficient for its PCR based downstream application, but unfortunately it can’t be used for whole genome sequencing or genotyping from extracted gDNA. Sequencing or genotyping can be achieved by after increasing template copy number through whole genome amplification of extracted gDNA. The obtained results create a base for future research to develop high-throughput research and extraction methods from blood samples. Conclusion: The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies.

scholarly journals A surrogate virus neutralization test to quantify antibody-mediated inhibition of SARS-CoV-2 in finger stick dried blood spot samples

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Amelia E. Sancilio ◽  
Richard T. D’Aquila ◽  
Elizabeth M. McNally ◽  
Matthew P. Velez ◽  
Michael G. Ison ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Venous Blood ◽  
Host Cells ◽  
Dried Blood Spot ◽  
Blood Collection ◽  
Widespread Application ◽  
Live Virus ◽  
Dilution Series ◽  
Wide Range ◽  
Blood Spot

AbstractThe spike protein of SARS-CoV-2 engages the human angiotensin-converting enzyme 2 (ACE2) receptor to enter host cells, and neutralizing antibodies are effective at blocking this interaction to prevent infection. Widespread application of this important marker of protective immunity is limited by logistical and technical challenges associated with live virus methods and venous blood collection. To address this gap, we validated an immunoassay-based method for quantifying neutralization of the spike-ACE2 interaction in a single drop of capillary whole blood, collected on filter paper as a dried blood spot (DBS) sample. Samples are eluted overnight and incubated in the presence of spike antigen and ACE2 in a 96-well solid phase plate. Competitive immunoassay with electrochemiluminescent label is used to quantify neutralizing activity. The following measures of assay performance were evaluated: dilution series of confirmed positive and negative samples, agreement with results from matched DBS-serum samples, analysis of results from DBS samples with known COVID-19 status, and precision (intra-assay percent coefficient of variation; %CV) and reliability (inter-assay; %CV). Dilution series produced the expected pattern of dose–response. Agreement between results from serum and DBS samples was high, with concordance correlation = 0.991. Analysis of three control samples across the measurement range indicated acceptable levels of precision and reliability. Median % surrogate neutralization was 46.9 for PCR confirmed convalescent COVID-19 samples and 0.1 for negative samples. Large-scale testing is important for quantifying neutralizing antibodies that can provide protection against COVID-19 in order to estimate the level of immunity in the general population. DBS provides a minimally-invasive, low cost alternative to venous blood collection, and this scalable immunoassay-based method for quantifying inhibition of the spike-ACE2 interaction can be used as a surrogate for virus-based assays to expand testing across a wide range of settings and populations.

Development and validation of a dried blood spot–HPLC assay for the determination of metronidazole in neonatal whole blood samples

2010 ◽  
Vol 397 (2) ◽  
pp. 687-693 ◽  
Author(s):  
Maysa Faisal Suyagh ◽  
Godwill Iheagwaram ◽  
Prashant Laxman Kole ◽  
Jeff Millership ◽  
Paul Collier ◽  
...  
Keyword(s):  
Whole Blood ◽  
Dried Blood Spot ◽  
Hplc Assay ◽  
Blood Samples ◽  
Whole Blood Samples ◽  
Development And Validation ◽  
Blood Spot

scholarly journals Dried Blood Spot Analysis Suitable for Therapeutic Drug Monitoring of Voriconazole, Fluconazole, and Posaconazole

2013 ◽  
Vol 57 (10) ◽  
pp. 4999-5004 ◽  
Author(s):  
Kim C. M. van der Elst ◽  
Lambert F. R. Span ◽  
Kai van Hateren ◽  
Karin M. Vermeulen ◽  
Tjip S. van der Werf ◽  
...  
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Drug Monitoring ◽  
Sampling Method ◽  
Fungal Infections ◽  
Venous Blood ◽  
Blood Sampling ◽  
Dried Blood Spot ◽  
Therapeutic Drug ◽  
Venous Blood Sampling ◽  
Blood Spot

ABSTRACTInvasive aspergillosis and candidemia are important causes of morbidity and mortality in immunocompromised and critically ill patients. The triazoles voriconazole, fluconazole, and posaconazole are widely used for the treatment and prophylaxis of these fungal infections. Due to the variability of the pharmacokinetics of the triazoles among and within individual patients, therapeutic drug monitoring is important for optimizing the efficacy and safety of antifungal treatment. A dried blood spot (DBS) analysis was developed and was clinically validated for voriconazole, fluconazole, and posaconazole in 28 patients. Furthermore, a questionnaire was administered to evaluate the patients' opinions of the sampling method. The DBS analytical method showed linearity over the concentration range measured for all triazoles. Results for accuracy and precision were within accepted ranges; samples were stable at room temperature for at least 12 days; and different hematocrit values and blood spot volumes had no significant influence. The ratio of the drug concentration in DBS samples to that in plasma was 1.0 for voriconazole and fluconazole and 0.9 for posaconazole. Sixty percent of the patients preferred DBS analysis as a sampling method; 15% preferred venous blood sampling; and 25% had no preferred method. There was significantly less perception of pain with the DBS sampling method (P= 0.021). In conclusion, DBS analysis is a reliable alternative to venous blood sampling and can be used for therapeutic drug monitoring of voriconazole, fluconazole, and posaconazole. Patients were satisfied with DBS sampling and had less pain than with venous sampling. Most patients preferred DBS sampling to venous blood sampling.

Evaluation of a novel dried blood spot collection device (HemaSpot™) to test blood samples collected from dogs for antibodies to Leishmania infantum

2014 ◽  
Vol 205 (1-2) ◽  
pp. 338-342 ◽  
Author(s):  
Alexa C. Rosypal ◽  
Leanne D. Pick ◽  
Jaime O. Esquivel Hernandez ◽  
David S. Lindsay
Keyword(s):  
Leishmania Infantum ◽  
Dried Blood Spot ◽  
Blood Samples ◽  
Collection Device ◽  
Blood Spot
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