Rapid detection of cancer-related cfc-DNA biomarkers directly from whole blood.

2012 ◽  
Vol 30 (30_suppl) ◽  
pp. 14-14
Author(s):  
Michael James Heller ◽  
Avery Sonnenberg
Keyword(s):  
Cancer Patient ◽  
Blood Sample ◽  
Whole Blood ◽  
High Field ◽  
Lymphocytic Leukemia ◽  
Clinical Samples ◽  
Diagnostic Systems ◽  
Serum Samples ◽  
Blood Samples ◽  
Green Fluorescent

14 Background: We have developed a “Sample to Answer” dielectrophoretic (DEP) technology for detecting cancer and other disease-related cfc-DNA biomarkers directly from whole blood. Using AC/DC electric field microarray devices specifically designed for the isolation of cfc-DNA, exosomes and cellular nanoparticulates from blood samples, experiments were carried out by adding about 20ul of the patient blood sample to the DEP microarray device. Methods: The DEP field was applied at 10 kHz and 20 Vp-p for 15 minutes to nine microelectrodes. The microarray was then washed three times with 0.5x PBS with the DEP field on, and finally examined by epifluorescent microscopy. Results: For Chronic Lymphocytic Leukemia (CLL) cancer patient blood samples, results clearly show greenish white fluorescent circles around the nine DEP microelectrodes on the array. This is the SYBR Green fluorescent stained cfc-DNA from the CLL cancer patient blood sample now concentrated in the DEP high field regions. Normal blood samples show little or no fluorescence around the DEP high field microelectrodes. More than fifty CLL samples have been analyzed to date. Conclusions: DEP devices are now being used to collect cfc-DNA from other cancer patient whole blood, plasma and serum samples. Blood sample to PCR is now achieved in less than twenty minutes. Newer work has demonstrated that PCR can be carried out in-situ (in the same device). Thus, the new DEP technology and devices set the stage for “seamless sample to answer” diagnostic systems which will allow a variety of important cancer and other disease biomarkers to be rapidly isolated and analyzed directly from whole blood and other clinical samples.

scholarly journals Tranexamic acid quantification in human whole blood using liquid samples or volumetric absorptive microsampling devices

Bioanalysis ◽  
2020 ◽  
Author(s):  
Elodie Lamy ◽  
Ileana Runge ◽  
Ian Roberts ◽  
Haleema Shakur-Still ◽  
Stanislas Grassin-Delyle
Keyword(s):  
Tranexamic Acid ◽  
Whole Blood ◽  
International Association ◽  
Clinical Samples ◽  
European Medicines Agency ◽  
Therapeutic Drug ◽  
Blood Samples ◽  
Human Whole Blood ◽  
Liquid Samples ◽  
Volumetric Absorptive Microsampling

Background: Recent clinical trials demonstrate the benefits of the antifibrinolytic drug tranexamic acid but its pharmacokinetics remain to be investigated more in depth. Although pharmacokinetics studies are usually performed with plasma, volumetric absorptive microsampling devices allow us to analyze dried whole blood samples with several advantages. Materials & methods: High-sensitivity LC–MS/MS methods for the quantification of tranexamic acid in human whole blood using liquid samples or dry samples on volumetric absorptive microsampling devices were developed and validated based on International Association from Therapeutic Drug Monitoring and Clinical Toxicology, European Medicines Agency and US Food and Drug Administration guidance. Conclusion: The method performances were excellent across the range of clinically relevant concentrations. The stability of tranexamic acid in blood samples stored up to 1 month at +50°C was demonstrated. The methods’ suitability was confirmed with clinical samples.

scholarly journals Peculiarities of vanadium determination in whole blood by icp-ms

2014 ◽  
Vol 60 (1) ◽  
pp. 109-114 ◽  
Author(s):  
T.S. Ulanova ◽  
O.V. Gileva ◽  
E.V. Stenno ◽  
G.A. Veikhman
Keyword(s):  
Sample Preparation ◽  
Blood Sample ◽  
Whole Blood ◽  
Concentration Level ◽  
Blood Samples ◽  
Reference Concentration ◽  
Icp Ms ◽  
Vanadium Determination ◽  
The Town

The parameters of vanadium determination by ICP-MS in whole blood are presented. Conditions for blood sample preparation to reduce measure errors and to determine vanadium at the reference concentration level were optimized. The accuracy of the results is confirmed by analysis of standard blood samples Seronorm L1, L2 and L3. Vanadium mean in whole blood for the group of children from the town of Chusovoy (n=80) was 1.29±0.45 µg/L, and vanadium mean for grown – ups from the town of Chusovoy was 1.63±0.25 µg/L.

scholarly journals Droplet Digital PCR for the Detection of Plasmodium falciparum DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 478
Author(s):  
Elena Pomari ◽  
Ronaldo Silva ◽  
Lucia Moro ◽  
Giulia La Marca ◽  
Francesca Perandin ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Dna Analysis ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Clinical Samples ◽  
Clinical Parameters ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Blood Samples ◽  
Paired Samples

Background: The estimation of Plasmodium falciparum parasitaemia can vary according to the method used. Recently, droplet digital PCR (ddPCR) has been proposed as a promising approach in the molecular quantitation of Plasmodium, but its ability to predict the actual parasitaemia on clinical samples has not been largely investigated. Moreover, the possibility of applying the ddPCR-sensitive method to serum samples has never been explored. Methods: We used, for the first time, ddPCR on both blood and serum to detect the DNA of P. falciparum in 52 paired samples from 26 patients. ddPCR was compared with loop-mediated isothermal amplification (LAMP) and rtPCR. The correlation between the ddPCR results, microscopy, and clinical parameters was examined. Results: ddPCR and microscopy were found to be strongly correlated (ρ(26) = 0.83111, p < 0.0001) in blood. Samples deviating from the correlation were partially explained by clinical parameters. In serum samples, ddPCR revealed the best performance in detecting P. falciparum DNA, with 77% positive samples among malaria subjects. Conclusion: Absolute quantitation by ddPCR can be a flexible technique for Plasmodium detection, with potential application in the diagnosis of malaria. In particular, ddPCR is a powerful approach for Plasmodium DNA analysis on serum when blood samples are unavailable.

Washing-free Chemiluminescence Immunoassay for Rapid Detection of cTnI in Whole Blood Samples

2021 ◽  
Author(s):  
Huan Zhao ◽  
Enben Su ◽  
Li Huang ◽  
Yunfeng Zai ◽  
Yuan Liu ◽  
...  
Keyword(s):  
Whole Blood ◽  
Rapid Detection ◽  
Magnetic Particles ◽  
Troponin I ◽  
Sample Type ◽  
Serum Samples ◽  
Chemiluminescence Immunoassay ◽  
Blood Samples ◽  
Significant Difference ◽  
Whole Blood Samples

Abstract Background: Chemiluminescence immunoassay (CLIA) has always been a great challenge in detecting whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Results: In this scheme, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotin-conjugated cTnI antibody and detected by streptavidin/acridine aster-conjugated PCMS. After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the cTnI concentrations of the serum samples, plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at 0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62~5.67%. The assay was linear over the studied range of 0.01–50.00 ng/mL, and no hook effect was found when cTnI concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit (Abbott assay kit), the correlation coefficient was 0.9859.Conclusions: A washing-free chemiluminescence immunoassay (CLIA) was established for the rapid detection of cardiac troponin I (cTnI) in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and polychloromethylstyrene microspheres (PCMS) for signal amplification, which showed great potential in clinical application.

Design and Development of a Unit-Use Cartridge for Coagulation Testing in Whole Blood

1999 ◽  
Author(s):  
Rhonda Cheadle ◽  
Andy Maczuszenko ◽  
Cindra Widrig Opalsky
Keyword(s):  
Blood Sample ◽  
Whole Blood ◽  
Clot Formation ◽  
Coagulation Cascade ◽  
Design And Development ◽  
Blood Samples ◽  
Coagulation Assays ◽  
Coagulation Testing ◽  
Functional Features ◽  
Whole Blood Samples

Abstract The following describes the development of a disposable cartridge for use at the patient bedside to perform traditional coagulation assays on fresh whole blood samples. The cartridge provides a means by which a blood sample can be metered and quantitatively mixed with reagents that activate the coagulation cascade. Clot formation is subsequently detected using a microfabricated sensor housed within the cartridge. The functional features of the cartridge and sensor are described.

scholarly journals Flexible Micro Spring Array Device for High-Throughput Enrichment of Viable Circulating Tumor Cells

2014 ◽  
Vol 60 (2) ◽  
pp. 323-333 ◽  
Author(s):  
Ramdane A Harouaka ◽  
Ming-Da Zhou ◽  
Yin-Ting Yeh ◽  
Waleed J Khan ◽  
Avisnata Das ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Whole Blood ◽  
Ex Vivo ◽  
Cell Damage ◽  
Flexible Structures ◽  
Capture Efficiency ◽  
Clinical Samples ◽  
Blood Samples ◽  
Whole Blood Samples

Abstract BACKGROUND The dissemination of circulating tumor cells (CTCs) that cause metastases in distant organs accounts for the majority of cancer-related deaths. CTCs have been established as a cancer biomarker of known prognostic value. The enrichment of viable CTCs for ex vivo analysis could further improve cancer diagnosis and guide treatment selection. We designed a new flexible micro spring array (FMSA) device for the enrichment of viable CTCs independent of antigen expression. METHODS Unlike previous microfiltration devices, flexible structures at the micro scale minimize cell damage to preserve viability, while maximizing throughput to allow rapid enrichment directly from whole blood with no need for sample preprocessing. Device performance with respect to capture efficiency, enrichment against leukocytes, viability, and proliferability was characterized. CTCs and CTC microclusters were enriched from clinical samples obtained from breast, lung, and colorectal cancer patients. RESULTS The FMSA device enriched tumor cells with 90% capture efficiency, higher than 104 enrichment, and better than 80% viability from 7.5-mL whole blood samples in &lt;10 min on a 0.5-cm2 device. The FMSA detected at least 1 CTC in 16 out of 21 clinical samples (approximately 76%) compared to 4 out of 18 (approximately 22%) detected with the commercial CellSearch® system. There was no incidence of clogging in over 100 tested fresh whole blood samples. CONCLUSIONS The FMSA device provides a versatile platform capable of viable enrichment and analysis of CTCs from clinically relevant volumes of whole blood.

scholarly journals Next-Generation Sequencing to Detect Pathogens in Pediatric Febrile Neutropenia: A Single-Center Retrospective Study of 112 Cases

2021 ◽  
Author(s):  
Kazuhiro Horiba ◽  
Yuka Torii ◽  
Toshihiko Okumura ◽  
Suguru Takeuchi ◽  
Takako Suzuki ◽  
...  
Keyword(s):  
Cluster Analysis ◽  
Next Generation Sequencing ◽  
Blood Culture ◽  
Febrile Neutropenia ◽  
Clinical Samples ◽  
Next Generation ◽  
Serum Samples ◽  
Blood Samples ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

Abstract Background Febrile neutropenia (FN) is a frequent complication in immunocompromised patients. However, causative microorganisms are detected in only 10% of patients. This study aimed to detect the microorganisms that cause FN using next-generation sequencing (NGS) to idenjpgy the genome derived from pathogenic microorganisms in the bloodstream. Here, we implemented a metagenomic approach to comprehensively analyze microorganisms present in clinical samples from patients with FN. Methods FN is defined as 1) a neutrophil count &lt; 500/µL, and 2) fever ≥ 37.5 °C. Plasma/serum samples of 112 pediatric patients with FN, 10 patients with neutropenia without fever (NE), were sequenced by NGS and analyzed by a metagenomic pipeline PATHDET. Results The putative pathogens were detected by NGS in 5 of 10 patients with FN with positive for blood culture results, 15 of 87 patients (17%) with negative for blood culture results, and 3 of 8 patients with NE. Several bacteria that were common in the oral, skin, and gut flora were commonly detected in blood samples, suggesting translocation of the human microbiota to the bloodstream in the setting of neutropenia. The cluster analysis of the microbiota in blood samples using NGS demonstrated that the representative bacteria of each cluster was mostly consistent with the pathogens in each patient. Conclusions NGS technique has a great potential for detecting causative pathogens in patients with FN. Cluster analysis, which extracts characteristic microorganisms from a complex microbial population, may be effective to detect pathogens in minute quantities of microbiota, such as those from the bloodstream.

scholarly journals A high-throughput microfluidic nanoimmunoassay for detecting anti–SARS-CoV-2 antibodies in serum or ultralow-volume blood samples

2021 ◽  
Vol 118 (18) ◽  
pp. e2025289118
Author(s):  
Zoe Swank ◽  
Grégoire Michielin ◽  
Hon Ming Yip ◽  
Patrick Cohen ◽  
Diego O. Andrey ◽  
...  
Keyword(s):  
Human Serum ◽  
High Throughput ◽  
Whole Blood ◽  
Test Strip ◽  
Serum Samples ◽  
Blood Samples ◽  
Novel Technologies ◽  
Human Serum Samples ◽  
Biomarker Analysis ◽  
Glucose Test

Novel technologies are needed to facilitate large-scale detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) specific antibodies in human blood samples. Such technologies are essential to support seroprevalence studies and vaccine clinical trials, and to monitor quality and duration of immunity. We developed a microfluidic nanoimmunoassay (NIA) for the detection of anti–SARS-CoV-2 IgG antibodies in 1,024 samples per device. The method achieved a specificity of 100% and a sensitivity of 98% based on the analysis of 289 human serum samples. To eliminate the need for venipuncture, we developed low-cost, ultralow-volume whole blood sampling methods based on two commercial devices and repurposed a blood glucose test strip. The glucose test strip permits the collection, shipment, and analysis of 0.6 μL of whole blood easily obtainable from a simple finger prick. The NIA platform achieves high throughput, high sensitivity, and specificity based on the analysis of 289 human serum samples, and negligible reagent consumption. We furthermore demonstrate the possibility to combine NIA with decentralized and simple approaches to blood sample collection. We expect this technology to be applicable to current and future SARS-CoV-2 related serological studies and to protein biomarker analysis in general.

Aqueous and serum-based materials compared for use as simulated calibrators for three ionized calcium analyzers.

1986 ◽  
Vol 32 (8) ◽  
pp. 1548-1550 ◽  
Author(s):  
J Toffaletti ◽  
C Bird ◽  
C Berg ◽  
B Abrams
Keyword(s):  
Reference Material ◽  
Human Serum ◽  
Whole Blood ◽  
Calcium Ion ◽  
Ionized Calcium ◽  
Serum Samples ◽  
Blood Samples ◽  
Serum Pool ◽  
Accuracy And Precision ◽  
Whole Blood Samples

Abstract To determine if bias between different ionized calcium analyzers could be decreased, we analyzed 10 control fluids during a study in which ionized calcium was measured in more than 150 serum and whole-blood samples. After calibrating three ionized calcium analyzers (Radiometer ICA 1, Nova 8, and AVL 980) with the manufacturers' respective calibrators, we used the between-instrument differences of the control fluids to simulate recalibration of the analyzers during each analytical run. A filtered human serum pool containing ionized calcium at 1 mmol/L concentration, with CO2 removed and having no added buffer, was the only material that consistently decreased between-analyzer bias of both serum and whole blood. Another human serum pool containing about 1.3 mmol of ionized calcium and about 10 mmol of bicarbonate per liter was even better at minimizing analyzer biases for serum samples, but was not as effective for whole-blood samples. Some additives used to buffer pH apparently adversely affected both the accuracy and precision of some, but not other, calcium ion electrodes. We conclude that if a reference material is developed for calibration of ionized calcium analyzers, it should be tested on several analyzers for use with both serum and whole blood, and it should be at least as effective as a human serum material, such as that used here.

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