scholarly journals A high-throughput microfluidic nano-immunoassay for detecting anti-SARS-CoV-2 antibodies in serum or ultra-low volume dried blood samples

2020 ◽  
Author(s):  
Zoe Swank ◽  
Grégoire Michielin ◽  
Hon Ming Yip ◽  
Patrick Cohen ◽  
Diego O. Andrey ◽  
...  
Keyword(s):  
High Throughput ◽  
Whole Blood ◽  
Large Scale ◽  
Test Strip ◽  
Sample Collection ◽  
Serum Samples ◽  
Blood Samples ◽  
Novel Technologies ◽  
Glucose Test ◽  
Low Volume

AbstractNovel technologies are needed to facilitate large-scale detection and quantification of SARS-CoV-2 specific antibodies in human blood samples. Such technologies are essential to support seroprevalence studies, vaccine clinical trials, and to monitor quality and duration of immunity. We developed a microfluidic nano-immunnoassay for the detection of anti-SARS-CoV-2 IgG antibodies in 1024 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, ultra-low 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 whole blood easily obtainable from a simple fingerprick. The nano-immunoassay platform achieves high-throughput, high sensitivity and specificity, negligible reagent consumption, and a decentralized and simple approach to blood sample collection. We expect this technology to be immediately applicable to current and future SARS-CoV-2 related serological studies and to protein biomarker diagnostics in general.

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.

scholarly journals Feasibility of High-Throughput Genome-Wide Genotyping using DNA from Stored Buccal Cell Samples

2010 ◽  
Vol 5 ◽  
pp. BMI.S5062 ◽  
Author(s):  
Stephanie J. Loomis ◽  
Lana M. Olson ◽  
Louis R. Pasquale ◽  
Janey Wiggs ◽  
Daniel Mirel ◽  
...  
Keyword(s):  
High Throughput ◽  
Large Scale ◽  
Buccal Cell ◽  
Health Study ◽  
Attractive Alternative ◽  
Sample Collection ◽  
Blood Samples ◽  
Dna Concentration ◽  
Genome Wide ◽  
Cell Sample

It is unclear if buccal cell samples contain sufficient human DNA with adequately sized fragments for high throughput genetic bioassays. Yet buccal cell sample collection is an attractive alternative to gathering blood samples for genetic epidemiologists engaged in large-scale genetic biomarker studies. We assessed the genotyping efficiency (GE) and genotyping concordance (GC) of buccal cell DNA samples compared to corresponding blood DNA samples, from 32 Nurses' Health Study (NHS) participants using the Illumina Infinium 660W-Quad platform. We also assessed how GE and GC accuracy varied as a function of DNA concentration using serial dilutions of buccal DNA samples. Finally we determined the nature and genomic distribution of discordant genotypes in buccal DNA samples. The mean GE of undiluted buccal cell DNA samples was high (99.32%), as was the GC between the paired buccal and blood samples (99.29%). GC between the dilutions versus the undiluted buccal DNA was also very high (>97%), though both GE and GC notably declined at DNA concentrations less than 5 ng/μl. Most (>95%) genotype determinations in buccal cell samples were of the “missing call” variety (as opposed to the “alternative genotype call” variety) across the spectrum of buccal DNA concentrations studied. Finally, for buccal DNA concentration above 1.7 ng/ul, discordant genotyping calls did not cluster in any particular chromosome. Buccal cell-derived DNA represents a viable alternative to blood DNA for genotyping on a high-density platform.

scholarly journals Factors Enhancing Serum Syndecan-1 Concentrations: A Large-Scale Comprehensive Medical Examination

2019 ◽  
Vol 8 (9) ◽  
pp. 1320
Author(s):  
Kazumasa Oda ◽  
Hideshi Okada ◽  
Akio Suzuki ◽  
Hiroyuki Tomita ◽  
Ryo Kobayashi ◽  
...  
Keyword(s):  
Large Scale ◽  
Prospective Observational Study ◽  
Nitrogen Concentration ◽  
Laboratory Data ◽  
University Hospital ◽  
Endothelial Glycocalyx ◽  
Sample Collection ◽  
Blood Samples ◽  
Triglyceride Concentration ◽  
Medical Examinations

Endothelial disorders are related to various diseases. An initial endothelial injury is characterized by endothelial glycocalyx injury. We aimed to evaluate endothelial glycocalyx injury by measuring serum syndecan-1 concentrations in patients during comprehensive medical examinations. A single-center, prospective, observational study was conducted at Asahi University Hospital. The participants enrolled in this study were 1313 patients who underwent comprehensive medical examinations at Asahi University Hospital from January 2018 to June 2018. One patient undergoing hemodialysis was excluded from the study. At enrollment, blood samples were obtained, and study personnel collected demographic and clinical data. No treatments or exposures were conducted except for standard medical examinations and blood sample collection. Laboratory data were obtained by the collection of blood samples at the time of study enrolment. According to nonlinear regression, the concentrations of serum syndecan-1 were significantly related to age (p = 0.016), aspartic aminotransferase concentration (AST, p = 0.020), blood urea nitrogen concentration (BUN, p = 0.013), triglyceride concentration (p < 0.001), and hematocrit (p = 0.006). These relationships were independent associations. Endothelial glycocalyx injury, which is reflected by serum syndecan-1 concentrations, is related to age, hematocrit, AST concentration, BUN concentration, and triglyceride concentration.

scholarly journals A rapid high-throughput sequencing-based approach for the identification of unknown bacterial pathogens in whole blood

2020 ◽  
Vol 6 (6) ◽  
pp. FSO476
Author(s):  
Ofir Israeli ◽  
Efi Makdasi ◽  
Inbar Cohen-Gihon ◽  
Anat Zvi ◽  
Shirley Lazar ◽  
...  
Keyword(s):  
High Throughput ◽  
Whole Blood ◽  
High Throughput Sequencing ◽  
High Background ◽  
Blood Samples ◽  
Bacterial Enrichment ◽  
Host Pathogen ◽  
High Throughput Dna Sequencing ◽  
Human Blood Samples ◽  
Pathogen Dna

High-throughput DNA sequencing (HTS) of pathogens in whole blood samples is hampered by the high host/pathogen nucleic acids ratio. We describe a novel and rapid bacterial enrichment procedure whose implementation is exemplified in simulated bacteremic human blood samples. The procedure involves depletion of the host DNA, rapid HTS and bioinformatic analyses. Following this procedure, Y. pestis, F. tularensis and B. anthracis spiked-in samples displayed an improved host/pathogen DNA ratio of 2.5–5.9 orders of magnitude, in samples with bacteria spiked-in at 103–105 CFU/ml. The procedure described in this study enables rapid and detailed metagenomic profiling of pathogens within 8–9 h, circumventing the challenges imposed by the high background present in the bacteremic blood and by the unknown nature of the sample.

scholarly journals Measurement of Glycated Hemoglobin A1c from Dried Blood by Turbidimetric Immunoassay

2009 ◽  
Vol 3 (5) ◽  
pp. 1203-1206 ◽  
Author(s):  
Ramakrishnan Lakshmy ◽  
Ruby Gupta
Keyword(s):  
Whole Blood ◽  
Large Scale ◽  
Hemoglobin A1c ◽  
Glycated Hemoglobin ◽  
Intraclass Correlation ◽  
Dried Blood Spots ◽  
Sample Collection ◽  
Blood Spots ◽  
Glycated Hemoglobin A1c ◽  
The Impact

Background: Glycated hemoglobin A1c (A1C) is an important marker in the diagnosis and treatment of diabetes. Dried blood measurement of A1C is useful in large scale epidemiological evaluation of A1C, especially to assess the impact of intervention programs. The possibility of using dried blood for measurement of A1C by the immunoturbidimetric method was explored in the present study. Method: Blood was collected from 30 patients, and blood spots were prepared and dried. The dried blood spot samples were kept for different lengths of time at 4°C to assess stability. Glycated hemoglobin was measured in whole blood and dried blood on the day of collection as well as on days 10 and 15 by immunoturbidimetric method. Results: The A1C values of 30 samples analyzed for comparison between whole blood estimation and dried blood ranged from 4.6% to 9.9%. The mean A1C on the day of sample collection was 6.01% ± 1.58% in fresh whole blood samples and 5.94% ± 1.58 % in dried blood spots. A linear and highly correlated relationship was observed between dried blood A1C values and those in whole blood ( r = 0.986 and intraclass correlation value = 0.993). Glycated hemoglobin values on day 10 and day 15 were comparable with the values on day 1 with a shift in mean of just 1% on day 10 and 3.04% on day 15. Conclusion: In conclusion, dried blood can be used for measurement of A1C by immunoturbidimetric method, and further stability of A1C measurement from dried blood for up to 15 days at 4°C makes it an ideal matrix for transportation in developing countries like India.

scholarly journals A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Michal Kucharski ◽  
Jaishree Tripathi ◽  
Sourav Nayak ◽  
Lei Zhu ◽  
Grennady Wirjanata ◽  
...  
Keyword(s):  
High Throughput ◽  
Large Scale ◽  
Rna Extraction ◽  
Transcriptome Data ◽  
High Quality ◽  
Diverse Range ◽  
Human Haemoglobin ◽  
Infected Blood ◽  
Blood Samples ◽  
Total Rna

Abstract Background Sequencing technology advancements opened new opportunities to use transcriptomics for studying malaria pathology and epidemiology. Even though in recent years the study of whole parasite transcriptome proved to be essential in understanding parasite biology there is no compiled up-to-date reference protocol for the efficient generation of transcriptome data from growing number of samples. Here, a comprehensive methodology on how to preserve, extract, amplify, and sequence full-length mRNA transcripts from Plasmodium-infected blood samples is presented that can be fully streamlined for high-throughput studies. Results The utility of various commercially available RNA-preserving reagents in a range of storage conditions was evaluated. Similarly, several RNA extraction protocols were compared and the one most suitable method for the extraction of high-quality total RNA from low-parasitaemia and low-volume blood samples was established. Furthermore, the criteria needed to evaluate the quality and integrity of Plasmodium RNA in the presence of human RNA was updated. Optimization of SMART-seq2 amplification method to better suit AT-rich Plasmodium falciparum RNA samples allowed us to generate high-quality transcriptomes from as little as 10 ng of total RNA and a lower parasitaemia limit of 0.05%. Finally, a modified method for depletion of unwanted human haemoglobin transcripts using in vitro CRISPR-Cas9 treatment was designed, thus improving parasite transcriptome coverage in low parasitaemia samples. To prove the functionality of the pipeline for both laboratory and field strains, the highest  2-hour resolution RNA-seq transcriptome for P. falciparum 3D7 intraerythrocytic life cycle available to  date was generated, and the entire protocol was applied to create the largest transcriptome data from Southeast Asian field isolates. Conclusions Overall, the presented methodology is an inclusive pipeline for generation of good quality transcriptomic data from a diverse range of Plasmodium-infected blood samples with varying parasitaemia and RNA inputs. The flexibility of this pipeline to be adapted to robotic handling will facilitate both small and large-scale future transcriptomic studies in the field of malaria.

Use of blood specimens collected on filter paper in screening for abnormal hemoglobins.

1976 ◽  
Vol 22 (5) ◽  
pp. 685-687 ◽  
Author(s):  
R M Schmidt ◽  
E M Brosious ◽  
S Holland ◽  
J M Wright ◽  
G R Serjeant
Keyword(s):  
Disease Control ◽  
Cellulose Acetate ◽  
Filter Paper ◽  
Whole Blood ◽  
Sample Collection ◽  
Capillary Tubes ◽  
Cellulose Acetate Electrophoresis ◽  
Blood Samples ◽  
Blood Specimens ◽  
The U.S

Abstract Both cellulose acetate electrophoresis and citrate agar electrophoresis were performed on 834 blood samples collected on filter paper in Jamaica and shipped for testing to the National Hemoglobinopathy Standardization Laboratory at the U.S. National Center for Disease Control. Additionally, 30 blood samples collected locally were stored on filter paper, in microhematocrit capillary tubes, and as whole blood specimens; at selected times the samples were tested for stability to determine the best sample-collection technique for hemoglobin electrophoresis. Results were most nearly accurate when both cellulose acetate electrophoresis and citrate agar testing were used. The methods are easy to perform, but results are unreliable if the blood samples on filter paper are stored at 4 degrees C for longer than two weeks before they are tested.

Disposable glucose test strip for whole blood with integrated sensing/diffusion-limiting layer

2009 ◽  
Vol 55 (2) ◽  
pp. 544-550 ◽  
Author(s):  
Zhencheng Chen ◽  
Cheng Fang ◽  
Hongyan Wang ◽  
Jishan He
Keyword(s):  
Whole Blood ◽  
Test Strip ◽  
Glucose Test

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.

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.

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