scholarly journals Improvement of Bovine Pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers.

2021 ◽  
Author(s):  
Paula Melisa Favaro ◽  
Ana Molineri ◽  
Maria J. Dus Santos ◽  
Luis F. Calvinho ◽  
Andrea Pecora
Keyword(s):  
Sampling Method ◽  
High Sensitivity ◽  
Cost Effective ◽  
Economic Losses ◽  
Serum Samples ◽  
Cost Effective Method ◽  
Positive Serum ◽  
Causative Agents ◽  
Filter Papers ◽  
Test Serum

Abstract Bovine pestiviruses are the causative agents of Bovine viral diarrhea, a disease that generates severe economic losses in cattle. The aim of this study was to improve its diagnosis by means of developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set a protocol for collecting, shipping and conserving bovine pestiviral RNA in filter papers. The developed RT-qPCR showed high sensitivity in detecting these viruses in different matrixes: viral stocks, semen and serum samples. Regarding the possibility of using the technique to test serum pools, it was possible to identify a positive serum sample within a pool containing 30 sera. In addition to evaluating the qPCR from fresh samples, the use of filter papers to sow bovine samples was analyzed. The sampling method in two different filter papers using bovine blood drops was a useful alternative for diagnosis purposes and allowed the preservation of pestiviral RNA up to 12 months under refrigeration.

scholarly journals Computer-supported Detection of M-Components and Evaluation of Immunoglobulins after Capillary Electrophoresis

2001 ◽  
Vol 47 (1) ◽  
pp. 110-117 ◽  
Author(s):  
Magnus Jonsson ◽  
Joyce Carlson ◽  
Jan-Olof Jeppsson ◽  
Per Simonsson
Keyword(s):  
Capillary Electrophoresis ◽  
Decision Support ◽  
Protein Separation ◽  
Serum Proteins ◽  
Cost Effective ◽  
Clinical Samples ◽  
Serum Samples ◽  
Computerized Decision Support ◽  
Cost Effective Method ◽  
Mathematical Algorithms

Abstract Background: Electrophoresis of serum samples allows detection of monoclonal gammopathies indicative of multiple myeloma, Waldenström macroglobulinemia, monoclonal gammopathy of undetermined significance, and amyloidosis. Present methods of high-resolution agarose gel electrophoresis (HRAGE) and immunofixation electrophoresis (IFE) are manual and labor-intensive. Capillary zone electrophoresis (CZE) allows rapid automated protein separation and produces digital absorbance data, appropriate as input for a computerized decision support system. Methods: Using the Beckman Paragon CZE 2000 instrument, we analyzed 711 routine clinical samples, including 95 monoclonal components (MCs) and 9 cases of Bence Jones myeloma, in both the CZE and HRAGE systems. Mathematical algorithms developed for the detection of monoclonal immunoglobulins (MCs) in the γ- and β-regions of the electropherogram were tested on the entire material. Additional algorithms evaluating oligoclonality and polyclonal concentrations of immunoglobulins were also tested. Results: CZE electropherograms corresponded well with HRAGE. Only one IgG MC of 1 g/L, visible on HRAGE, was not visible after CZE. Algorithms detected 94 of 95 MCs (98.9%) and 100% of those visible after CZE. Of 607 samples lacking an MC on HRAGE, only 3 were identified by the algorithms (specificity, 99%). Algorithms evaluating total gammaglobulinemia and oligoclonality also identified several cases of Bence Jones myeloma. Conclusions: The use of capillary electrophoresis provides a modern, rapid, and cost-effective method of analyzing serum proteins. The additional option of computerized decision support, which provides rapid and standardized interpretations, should increase the clinical availability and usefulness of protein analyses in the future.

scholarly journals Coupling multiplex pre-amplification and droplet digital PCR for longitudinal monitoring ofESR1andPIK3CAmutations from plasma cell-free DNA

2019 ◽  
Author(s):  
Huilan Yao ◽  
Grant Wu ◽  
Subhasree Das ◽  
Crystal MacKenzie ◽  
Hua Gao ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Hot Spot ◽  
Metastatic Breast ◽  
High Sensitivity ◽  
Cost Effective ◽  
Digital Pcr ◽  
Cost Effective Method ◽  
High Prevalence ◽  
High Concordance ◽  
Hotspot Mutations

AbstractHere we report on the development of a sensitive and cost-effective method to longitudinally trackESR1andPIK3CAmutations from cfDNA in patients with metastatic breast cancer (MBC) using a streamlined and de-centralized workflow. Hotspot mutations inESR1have been shown to cause resistance to aromatase inhibitor–based and anti-estrogenic therapies, whilePIK3CAmutations have high prevalence in MBC. As a result, their utility as circulating biomarkers to predict or monitor response in the clinical development of investigational compounds has been the focus of many studies. Six regions inESR1andPIK3CAgenes containing 20 hotspot mutations were pre-amplified, followed by optimized singleplex ddPCR assays to detect allele frequencies of individual mutations. Without pre-amplification, the limit of detection (LOD) and limit of linearity (LOL) of individual ddPCR assays were at 0.05-0.1% and 0.25% level, respectively. With pre-amplification, the LOD and LOL were slightly elevated at 0.1-0.25% and 0.25-0.5% levels, respectively. High concordance was achieved to the BEAMing assay (Sysmex Inostics) for mutation positive assays (r=0.98, P<0.0001). In conclusion, coupling pre-amplification and ddPCR assays allowed us for the detection of up to 20 hot spot mutations inESR1andPIK3CAwith high sensitivity and reproducibility.

scholarly journals Fabrication and Characterization of Humidity Sensors Based on Graphene Oxide–PEDOT:PSS Composites on a Flexible Substrate

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 148 ◽  
Author(s):  
Francisco J. Romero ◽  
Almudena Rivadeneyra ◽  
Markus Becherer ◽  
Diego P. Morales ◽  
Noel Rodríguez
Keyword(s):  
Graphene Oxide ◽  
Large Scale ◽  
Flexible Substrate ◽  
High Sensitivity ◽  
Cost Effective ◽  
Humidity Sensors ◽  
Good Thermal Stability ◽  
Cost Effective Method ◽  
And Performance ◽  
Stability Time

In this paper, we present a simple, fast, and cost-effective method for the large-scale fabrication of high-sensitivity humidity sensors on flexible substrates. These sensors consist of a micro screen-printed capacitive structure upon which a sensitive layer is deposited. We studied two different structures and three different sensing materials by modifying the concentration of poly(3,4-ethylenedioxythiophene)/polystyrene sulfonate (PEDOT:PSS) in a graphene oxide (GO) solution. The results show that the aggregation of the PEDOT:PSS to the GO can modify its electrical properties, boosting the performance of the capacitive sensors in terms of both resistive losses and sensitivity to relative humidity (RH) changes. Thus, in an area less than 30 mm2, the GO/PEDOT:PSS-based sensors can achieve a sensitivity much higher (1.22 nF/%RH at 1 kHz) than other similar sensors presented in the literature which, together with their good thermal stability, time response, and performance over bending, demonstrates that the manufacturing approach described in this work paves the way for the mass production of flexible humidity sensors in an inexpensive way.

scholarly journals High-Quality, Cost-Effective Strategy for Detection of Autoantibodies to Extractable Nuclear Antigens

2001 ◽  
Vol 8 (3) ◽  
pp. 471-474 ◽  
Author(s):  
Tri G. Phan ◽  
Watson W. S. Ng ◽  
Dana Bird ◽  
Kara Smithers ◽  
Vicky Wong ◽  
...  
Keyword(s):  
Cost Effective ◽  
Turnaround Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Quality Cost ◽  
Nuclear Antigens ◽  
Cost Effective Method ◽  
Meaningful Result ◽  
Clinical Diagnoses

ABSTRACT We evaluated methods for the detection of autoantibodies to extractable nuclear antigens (ENAs) to determine the strategy that yielded the most cost effective and clinically meaningful result. We prospectively compared counterimmunoelectrophoresis (CIEP) with and without serum prediffusion (SPD) and found that SPD significantly improved the quality of precipitation lines. This resulted in a decreased requirement for repeat testing and, consequently, was associated with a significant decrease in reagent costs and specimen turnaround time. We also retrospectively compared reactivity by CIEP, CIEP plus SPD, enzyme-linked immunosorbent assay (ELISA), and line immunoassay (LIA) of 52 serum samples that were previously determined to be positive for ENAs, and we correlated the results with clinical diagnoses. There was significant agreement among CIEP, CIEP plus SPD, ELISA, and LIA for the detection of anti-SS-A, anti-SS-B and anti-RNP. In general, CIEP, CIEP plus SPD, and LIA correlated better with the clinical diagnoses than ELISA, even though ELISA detected anti-ENAs more often than the other methods. CIEP plus SPD is therefore the most cost effective method for the identification of clinically meaningful ENAs. Based on our experience, we now screen for ENAs by CIEP, and positive samples are then typed by CIEP plus SPD. Samples that are difficult to interpret are then further assessed by an alternative method.

scholarly journals STAMP: a multiplex sequencing method for simultaneous evaluation of mitochondrial DNA heteroplasmies and content

2020 ◽  
Author(s):  
Xiaoxian Guo ◽  
Yiqin Wang ◽  
Ruoyu Zhang ◽  
Zhenglong Gu
Keyword(s):  
Large Scale ◽  
Nuclear Dna ◽  
Large Population ◽  
High Sensitivity ◽  
Cost Effective ◽  
Cost Effective Method ◽  
Sequencing Method ◽  
Scale Population ◽  
Mtdna Variants ◽  
Multiplex Sequencing

ABSTRACTHuman mitochondrial genome (mtDNA) variations, such as mtDNA heteroplasmies (the co-existence of mutated and wild-type mtDNA), have received increasing attention in recent years for their clinical relevance to numerous diseases. But large-scale population studies of mtDNA heteroplasmies have been lagging due to the lack of a labor- and cost-effective method. Here, we present a novel human mtDNA sequencing method called STAMP (sequencing by targeted amplification of multiplex probes) for measuring mtDNA heteroplasmies and content in a streamlined workflow. We show that STAMP has high mapping rates to mtDNA, deep coverage of unique reads, and high tolerance to sequencing and PCR errors when applied to human samples. STAMP also has high sensitivity and low false positive rates in identifying artificial mtDNA variants at fractions as low as 0.5% in genomic DNA samples. We further extend STAMP, by including nuclear DNA-targeting probes, to enable assessment of relative mtDNA content in the same assay. The high cost-effectiveness of STAMP, along with the flexibility of using it for measuring various aspects of mtDNA variations, will accelerate the research of mtDNA heteroplasmies and content in large population cohorts, and in the context of human diseases and aging.

scholarly journals Application of insecticides in industrial animal breeding

2019 ◽  
pp. 102-107
Author(s):  
A. P. Paliy ◽  
A. M. Mashkey ◽  
N. V. Sumakova ◽  
V. V. Gontar ◽  
A. P. Paliy
Keyword(s):  
Large Range ◽  
Practical Importance ◽  
Animal Husbandry ◽  
Cost Effective ◽  
Farm Animals ◽  
Economic Losses ◽  
Animal Products ◽  
Resistance To Insects ◽  
Modern Methods ◽  
Causative Agents

Entomoses of farm animals are widespread in the territory of Ukraine and cause significant economic losses to animal husbandry. It is established that the sick animals have reduced milk, meat and wool productivity, breeding qualities; weakened young animals, which are easily exposed to various diseases of infectious and non-infectious etiology, are born. Among all modern methods and means for artificial reduction of the number of insects, the most effective is the chemical method. To protect animals from midges the most cost-effective is the spraying of animals with insecticides and repellents. The analysis of the presented literature data allows us to say that sufficiently large range of effective preparations of both domestic and foreign production is presented on the market of disinsection agents. However, it has been reported that resistance to insects has formed for most of them, some of the products are highly toxic to warm-blooded animals, and also they are quite expensive and their use is economically unjustified. Great scientific and practical importance has the development of modern methods of combating the causative agents of farm animal entomoses based on strict regulations for treatment-and-prophylactic means, which make it possible to reduce the number of parasites to an economically intangible level, prevent environmental pollution by pesticides, and obtain safe animal products of high sanitary quality. The insecticide market has a fairly large range of efficient products, both domestic and foreign, but most of them do not meet modern challenges and advanced livestock technologies. At the present stage of the disinfectology development, the search for new compositions of chemical compounds for disinsection in animal husbandry to combat harmful insects is promising

scholarly journals STAMP: a multiplex sequencing method for simultaneous evaluation of mitochondrial DNA heteroplasmies and content

2020 ◽  
Vol 2 (4) ◽  
Author(s):  
Xiaoxian Guo ◽  
Yiqin Wang ◽  
Ruoyu Zhang ◽  
Zhenglong Gu
Keyword(s):  
Large Scale ◽  
Nuclear Dna ◽  
Large Population ◽  
High Sensitivity ◽  
Cost Effective ◽  
Cost Effective Method ◽  
Sequencing Method ◽  
Scale Population ◽  
Mtdna Variants ◽  
Multiplex Sequencing

Abstract Human mitochondrial genome (mtDNA) variations, such as mtDNA heteroplasmies (the co-existence of mutated and wild-type mtDNA), have received increasing attention in recent years for their clinical relevance to numerous diseases. But large-scale population studies of mtDNA heteroplasmies have been lagging due to the lack of a labor- and cost-effective method. Here, we present a novel human mtDNA sequencing method called STAMP (sequencing by targeted amplification of multiplex probes) for measuring mtDNA heteroplasmies and content in a streamlined workflow. We show that STAMP has high-mapping rates to mtDNA, deep coverage of unique reads and high tolerance to sequencing and polymerase chain reaction errors when applied to human samples. STAMP also has high sensitivity and low false positive rates in identifying artificial mtDNA variants at fractions as low as 0.5% in genomic DNA samples. We further extend STAMP, by including nuclear DNA-targeting probes, to enable assessment of relative mtDNA content in the same assay. The high cost-effectiveness of STAMP, along with the flexibility of using it for measuring various aspects of mtDNA variations, will accelerate the research of mtDNA heteroplasmies and content in large population cohorts, and in the context of human diseases and aging.

scholarly journals A Simple Colorimetric Molecular Detection of Novel Coronavirus (COVID-19), an Essential Diagnostic Tool for Pandemic Screening

2020 ◽  
Author(s):  
Pazhanimuthu Annamalai ◽  
Madhu Kanta ◽  
Pazhanivel Ramu ◽  
Baskar Ravi ◽  
Kokilavani Veerapandian ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Point Of Care ◽  
Lamp Assay ◽  
High Sensitivity ◽  
Cost Effective ◽  
Cost Effective Method ◽  
Recent Outbreak ◽  
Pcr Method ◽  
Novel Coronavirus ◽  
Virus Isolates

AbstractThe recent outbreak of the newly emerged novel coronavirus (SARS-CoV-2) presents a big challenge for public health laboratories as virus isolates are not available while there is an increasing evidence that the epidemic is more widespread than initially thought, as well as spreading internationally across borders through travellers does already happen warranting a methodology for the rapid detection of the infection to control SARS-CoV-2. Aim: We intended to develop and deploy a robust and rapid diagnostic methodology using LAMP assay for use in point of care settings to detect SARS-COV-2 infection. Methodology: In the present study, we have developed a validated rapid diagnostic procedure to detect SARS-CoV-2 using LAMP assay, its design relying on isothermal amplification of the nucleic acids of the SARS-CoV-2. Results: The LAMP assay developed detects SARS-CoV-2 infection rapidly with high sensitivity and reliability. The data generated by LAMP assay were comparable and at par with the data generated by real-time PCR method. Conclusion: The present study demonstrates that the LAMP assay developed was a rapid, reliable, sensitive and cost effective method to detect SARS-CoV-2 infection in a point of care as well as in laboratory settings.

scholarly journals Deep Learning-Enabled Point-of-Care Sensing Using Multiplexed Paper-Based Sensors

2019 ◽  
Author(s):  
Zachary Ballard ◽  
Hyou-Arm Joung ◽  
Artem Goncharov ◽  
Jesse Liang ◽  
Karina Nugroho ◽  
...  
Keyword(s):  
Machine Learning ◽  
Deep Learning ◽  
Point Of Care ◽  
Low Cost ◽  
High Sensitivity ◽  
Cost Effective ◽  
Serum Samples ◽  
Cvd Risk ◽  
Medical Test ◽  
Sensing Membrane

ABSTRACTWe present a deep learning-based framework to design and quantify point-of-care sensors. As its proof-of-concept and use-case, we demonstrated a low-cost and rapid paper-based vertical flow assay (VFA) for high sensitivity C-Reactive Protein (hsCRP) testing, a common medical test used for quantifying the degree of inflammation in patients at risk of cardio-vascular disease (CVD). A machine learning-based sensor design framework was developed for two key tasks: (1) to determine an optimal configuration of immunoreaction spots and conditions, spatially-multiplexed on a paper-based sensing membrane, and (2) to accurately infer the target analyte concentration based on the signals of the optimal VFA configuration. Using a custom-designed mobile-phone based VFA reader, a clinical study was performed with 85 human serum samples to characterize the quantification accuracy around the clinically defined cutoffs for CVD risk stratification. Results from blindly-tested VFAs indicate a competitive coefficient of variation of 11.2% with a linearity of R2 = 0.95; in addition to the success in the high-sensitivity CRP range (i.e., 0-10 mg/L), our results further demonstrate a mitigation of the hook-effect at higher CRP concentrations due to the incorporation of antigen capture spots within the multiplexed sensing membrane of the VFA. This paper-based computational VFA that is powered by deep learning could expand access to CVD health screening, and the presented machine learning-enabled sensing framework can be broadly used to design cost-effective and mobile sensors for various point-of-care diagnostics applications.

scholarly journals DIAGNOSTIC ACCURACY OF SALINE HYSTEROSONOGRAPHY IN DETECTING ENDOMETRIAL HYPERPLASIA IN PATIENTS WITH POST MENOPAUSAL BLEEDING

2020 ◽  
Vol 11 (2) ◽  
pp. 1-8
Author(s):  
Benish Yousuf ◽  
Hira Ambreen ◽  
Tahira Mariam ◽  
Abdul Raouf ◽  
Ambreen Yaseen ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Predictive Value ◽  
Endometrial Hyperplasia ◽  
Foley Catheter ◽  
High Sensitivity ◽  
Cost Effective ◽  
Postmenopausal Bleeding ◽  
Cost Effective Method ◽  
Post Menopausal ◽  
Sensitivity Specificity

BACKGROUND & OBJECTIVE: Saline hysterosonography is a simple and cost-effective method with high sensitivity to detect uterine abnormalities causing postmenopausal bleeding. The objective of this study was to evaluate the diagnostic accuracy of saline hysterosonography in detecting endometrial hyperplasia in women with postmenopausal bleeding by taking histopathology as a gold standard. METHODOLOGY: A hundred and twenty (120) cases were enrolled from the outpatient and inpatient department of obstetrics and gynecology. Proper history and relevant examination of the patient was done. Then preparations were made for the procedure. The patient was counseled and the technique explained to her. Then Foley catheter no 12 was passed in cervix and sonography was done while instilling normal saline through a cervical catheter and scan pictures were frozen and results were given by expert gynecologist of Allied Hospital, Faisalabad. Histopathology specimen was sent to the pathology lab. RESULTS: Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of saline hysterosonography in detecting endometrial hyperplasia was recorded as 96.15%,91,49%,75.76%,98.85%, and 92.5% respectively. CONCLUSION: Saline hysterosonography has high sensitivity to detect uterine hyperplasia. It can be used as a cost effective alternative to hysteroscopy in many units in Pakistan.

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