scholarly journals Modernizing the Toolkit for Arthropod Bloodmeal Identification

Insects ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 37
Author(s):  
Erin M. Borland ◽  
Rebekah C. Kading
Keyword(s):  
Nuclear Dna ◽  
Point Of Care ◽  
Isotope Analysis ◽  
Digital Pcr ◽  
Lessons Learned ◽  
Molecular Techniques ◽  
Melting Analysis ◽  
Field Deployment ◽  
Blood Meal Identification ◽  
Bloodmeal Identification

Understanding vertebrate–vector interactions is vitally important for understanding the transmission dynamics of arthropod-vectored pathogens and depends on the ability to accurately identify the vertebrate source of blood-engorged arthropods in field collections using molecular methods. A decade ago, molecular techniques being applied to arthropod blood meal identification were thoroughly reviewed, but there have been significant advancements in the techniques and technologies available since that time. This review highlights the available diagnostic markers in mitochondrial and nuclear DNA and discusses their benefits and shortcomings for use in molecular identification assays. Advances in real-time PCR, high resolution melting analysis, digital PCR, next generation sequencing, microsphere assays, mass spectrometry, and stable isotope analysis each offer novel approaches and advantages to bloodmeal analysis that have gained traction in the field. New, field-forward technologies and platforms have also come into use that offer promising solutions for point-of-care and remote field deployment for rapid bloodmeal source identification. Some of the lessons learned over the last decade, particularly in the fields of DNA barcoding and sequence analysis, are discussed. Though many advancements have been made, technical challenges remain concerning the prevention of sample degradation both by the arthropod before the sample has been obtained and during storage. This review provides a roadmap and guide for those considering modern techniques for arthropod bloodmeal identification and reviews how advances in molecular technology over the past decade have been applied in this unique biomedical context.

scholarly journals Integrated Extreme Real-Time PCR and High-Speed Melting Analysis in 52 to 87 Seconds

2019 ◽  
Vol 65 (2) ◽  
pp. 263-271 ◽  
Author(s):  
Joseph T Myrick ◽  
Robert J Pryor ◽  
Robert A Palais ◽  
Sean J Ison ◽  
Lindsay Sanford ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
High Speed ◽  
Dna Analysis ◽  
Point Of Care ◽  
Turnaround Time ◽  
Single Nucleotide Variants ◽  
Cycle Times ◽  
Pcr Products ◽  
Melting Analysis

Abstract BACKGROUND Extreme PCR in <30 s and high-speed melting of PCR products in <5 s are recent advances in the turnaround time of DNA analysis. Previously, these steps had been performed on different specialized instruments. Integration of both extreme PCR and high-speed melting with real-time fluorescence monitoring for detection and genotyping is presented here. METHODS A microfluidic platform was enhanced for speed using cycle times as fast as 1.05 s between 66.4 °C and 93.7 °C, with end point melting rates of 8 °C/s. Primer and polymerase concentrations were increased to allow short cycle times. Synthetic sequences were used to amplify fragments of hepatitis B virus (70 bp) and Clostridium difficile (83 bp) by real-time PCR and high-speed melting on the same instrument. A blinded genotyping study of 30 human genomic samples at F2 c.*97, F5 c.1601, MTHFR c.665, and MTHFR c.1286 was also performed. RESULTS Standard rapid-cycle PCR chemistry did not produce any product when total cycling times were reduced to <1 min. However, efficient amplification was possible with increased primer (5 μmol/L) and polymerase (0.45 U/μL) concentrations. Infectious targets were amplified and identified in 52 to 71 s. Real-time PCR and genotyping of single-nucleotide variants from human DNA was achieved in 75 to 87 s and was 100% concordant to known genotypes. CONCLUSIONS Extreme PCR with high-speed melting can be performed in about 1 min. The integration of extreme PCR and high-speed melting shows that future molecular assays at the point of care for identification, quantification, and variant typing are feasible.

Academic Family Health Teams Collaborative Lessons from the Far Flung North

2010 ◽  
pp. 350-369
Author(s):  
David Topps
Keyword(s):  
Point Of Care ◽  
Family Health ◽  
Data Access ◽  
Software Tools ◽  
Resource Discovery ◽  
Lessons Learned ◽  
Northern Ontario ◽  
Team Collaboration ◽  
Informatics Infrastructure ◽  
Family Health Team

Working collaboratively, in widely distributed settings, poses unique challenges. The Academic Family Health Team, affiliated with the Northern Ontario School of Medicine, has had to adopt a wide variety of information sharing practices and collaborative software tools, in order to function effectively in such roles as clinicians, educators and researchers. Based on an ongoing action research model, this chapter describes approaches taken and lessons learned while developing the informatics infrastructure to support interprofessional practice. The author describes how common procedures and software tools can benefit from a Web 2.0 approach, comparing commercial and open-source aspects of possible solutions. Ubiquitous data access for point of care decision making is supported by integrating web services, mobile devices and multi-stream communications. Resource discovery is enabled by integrating information streams into the medical record, into wireless device interfaces and via clinical dashboards. Effective team collaboration is highly enhanced through such infrastructure support.

scholarly journals Comparative Use of Quantitative PCR (qPCR), Droplet Digital PCR (ddPCR), and Recombinase Polymerase Amplification (RPA) in the Detection of Shiga Toxin-Producing E. coli (STEC) in Environmental Samples

Water ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3507
Author(s):  
Mark A. Ibekwe ◽  
Shelton E. Murinda ◽  
Stanley Park ◽  
Amarachukwu Obayiuwana ◽  
Marcia A. Murry ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Environmental Samples ◽  
Point Of Care ◽  
Foodborne Pathogen ◽  
Digital Pcr ◽  
Absolute Quantification ◽  
Droplet Digital Pcr ◽  
Recombinase Polymerase Amplification ◽  
High Rate ◽  
E Coli

E. coli O157:H7 is a foodborne pathogen that constitutes a global threat to human health. However, the quantification of this pathogen in food and environmental samples may be problematic at the low cell numbers commonly encountered in environmental samples. In this study, we used recombinase polymerase amplification (RPA) for the detection of E. coli O157:H7, real-time quantitative PCR (qPCR) for quantification, and droplet digital PCR (ddPCR) for absolute and accurate quantification of E. coli O157:H7 from spiked and environmental samples. Primer and probe sets were used for the detection of stx1 and stx2 using RPA. Genes encoding for stx1, stx2, eae, and rfbE were used to quantify E. coli O157:H7 in the water samples. Furthermore, duplex ddPCR assays were used to quantify the pathogens in these samples. Duplex assay set 1 used stx1 and rfbE genes, while assay set 2 used stx2 and eae genes. Droplet digital PCR was used for the absolute quantification of E. coli O15:H7 in comparison with qPCR for the spiked and environmental samples. The RPA results were compared to those from qPCR and ddPCR in order to assess the efficiency of the RPA compared with the PCR methods. The assays were further applied to the dairy lagoon effluent (DLE) and the high rate algae pond (HRAP) effluent, which were fed with diluted DLE. The RPA detected was <10 CFU/mL, while ddPCR showed quantification from 1 to 104 CFU/mL with a high reproducibility. In addition, quantification by qPCR was from 103 to 107 CFU/mL of the wastewater samples. Therefore, the RPA assay has potential as a point of care tool for the detection of E. coli O157:H7 from different environmental sources, followed by quantification of the target concentrations.

New Perspectives for Modern Trauma Management – Lessons Learned from 25 Years FAST and 15 Years E-FAST

2019 ◽  
Vol 40 (05) ◽  
pp. 560-583 ◽  
Author(s):  
Joseph Osterwalder ◽  
Gebhard Mathis ◽  
Beatrice Hoffmann
Keyword(s):  
Life Support ◽  
Point Of Care ◽  
Clinical Situation ◽  
Lessons Learned ◽  
Advanced Trauma Life Support ◽  
Point Of Care Ultrasound ◽  
Trauma Management ◽  
Abdominal Image ◽  
Trauma Surgeons ◽  
Ultrasound Applications

AbstractE-FAST (Extended-Focused Assessment with Sonography for Trauma) is now a widely utilized and internationally recognized standard exam in trauma care. It is highly accepted by emergency physicians and trauma surgeons alike. Thanks to the popularity of PoCUS (point-of-care ultrasound), it has continued to evolve over the last years and can now improve trauma diagnosis at all stages of the primary ABCDE. This review article summarizes key observations made over recent years and also highlights the extension of FAST into E-FAST in the context of PoCUS and CT developments for modern trauma management. Time has come to learn the lessons from 25 years of FAST and 15 years of E-FAST. We should redefine and position ultrasound in the primary ATLS survey (Advanced Trauma Life Support) on two levels: 1. Basic ATLS with new clinical questions, six additional abdominal image sections and one or more follow-up examinations depending on the clinical situation, and 2. Advanced ATLS with ultrasound applications for the entire trauma ABCDE.

DNA-DNA Hybridization, Phylogenetic Reconstruction and the Fossil Record

1988 ◽  
Vol 1 ◽  
pp. 75-88 ◽  
Author(s):  
Charles R. Marshall
Keyword(s):  
Dna Hybridization ◽  
Nuclear Dna ◽  
Phylogenetic Reconstruction ◽  
Cost Effective ◽  
Single Copy ◽  
Molecular Techniques ◽  
Phylogenetic Information ◽  
Molecular Approaches ◽  
Molecular Characters ◽  
Rates Of Molecular Evolution

In 1962 Zuckerkandl & Pauling suggested that the amino acid sequence of proteins might evolve in a clock-like fashion and thus may be useful for phylogenetic reconstruction. Since then, many different molecular approaches to phylogenetic reconstruction have been proposed (Wilson et al., 1977). Enthusiasm for the clock hypothesis was dampened by the discovery that rates of molecular evolution for many macromolecules have been highly variable through time (Romero-Herrera et al., 1979). However, the contribution of molecular characters to the study of phylogeny is not necessarily dependent on the notion of a molecular clock and molecular approaches continue to be an important source of phylogenetic information. One of the more powerful and cost-effective molecular techniques for phylogenetic purposes is DNA-DNA hybridization, which measures the single-copy nuclear DNA sequence divergences between species.

scholarly journals Development of the British Antarctic Survey Rapid Access Isotope Drill

2019 ◽  
Vol 65 (250) ◽  
pp. 288-298 ◽  
Author(s):  
JULIUS RIX ◽  
ROBERT MULVANEY ◽  
JIALIN HONG ◽  
DAN ASHURST
Keyword(s):  
Ice Core ◽  
Isotope Analysis ◽  
Lessons Learned ◽  
New Class ◽  
Future Developments ◽  
British Antarctic Survey ◽  
Rapid Access ◽  
Current State ◽  
Water Stable Isotope ◽  
Drilling Time

ABSTRACTThe British Antarctic Survey Rapid Access Isotope Drill is an innovative new class of electromechanical ice drill, which has recently been used to drill the deepest dry hole drilled by an electromechanical auger drill. The record-breaking depth of 461.58 m was drilled in just over 104 hours at Little Dome C. The drill collects ice chippings, for water stable isotope analysis, rather than an ice core. By not collecting a core the winch can be geared for speed rather than core breaking and is lightweight. Furthermore, emptying of the chippings is performed by simply reversing the drill motor on the surface reducing the overall drilling time significantly. The borehole is then available for instrumentation. We describe the drill in its current state including modifications carried out since it was last deployed. Test seasons and the lessons learned from each are outlined. Finally, future developments for this class of drill are discussed.

Initial Experience with Cancer Guidelines Navigator, a tool to standardize and improve the quality of cancer care in Sub-Saharan Africa, at Ocean Road Cancer Institute in Tanzania.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e14106-e14106
Author(s):  
Fernando Jose Suarez Saiz ◽  
Leemor Yuravlivker ◽  
Jerry Ndumbalo ◽  
Julius Mwaiselage ◽  
Sadiq Maalim Siu ◽  
...  
Keyword(s):  
Cancer Treatment ◽  
Treatment Planning ◽  
Reference System ◽  
Point Of Care ◽  
Lessons Learned ◽  
Ease Of Use ◽  
Sub Saharan Africa ◽  
Digital Reference ◽  
Sub Saharan ◽  
Cancer Guidelines

e14106 Background: The IBM Cancer Guidelines Navigator (CGN) is a digital reference system to support treatment planning that allows clinicians to enter a cancer patient’s clinical characteristics and presents the corresponding treatment options in the NCCN Harmonized Guidelines (TM) for Sub-Saharan Africa. In October 2019, Ocean Road Cancer Institute (ORCI) in Tanzania became the first site in Africa to initiate a hospital-wide implementation of the tool to help clinicians reduce cancer treatment variability by increasing adherence to standard evidence-based care. We describe training and lessons learned from system introduction. Methods: Training for clinical staff at ORCI occurred over one week and included daily one-hour lectures, followed by personalized hands-on training. A survey was administered to assess usability and use cases of the tool. Results: Thirty-one ORCI clinical and IT staff members participated in training, and 12 completed the survey. Responses indicated that the most beneficial uses for CGN were at point of care and for self-learning. Participants indicated that the top benefits of the tool were quick access to guidelines and evidence (75%) and ease of use (58%). Expanding cancer coverage (42%), offline access and better integration into the workflow (25%) were identified as areas for improvement. Post-training, ORCI implemented easier access to CGN on each computer and tablet used for consultation and care management. Conclusions: CGN is a digital reference system that is designed to support easy and efficient access to regionalized cancer-treatment guidelines for point-of-care treatment planning and education. Expansion of this program has been planned for other hospitals in Tanzania. Future studies will examine whether CGN usage affects guideline adherence.

scholarly journals Point-of-Care Testing as an Influenza Surveillance Tool: Methodology and Lessons Learned from Implementation

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Lisa H. Gren ◽  
Christina A. Porucznik ◽  
Elizabeth A. Joy ◽  
Joseph L. Lyon ◽  
Catherine J. Staes ◽  
...  
Keyword(s):  
Data Collection ◽  
Disease Surveillance ◽  
Point Of Care ◽  
Lessons Learned ◽  
Research Network ◽  
Influenza Surveillance ◽  
Statistical Process ◽  
University Of Utah ◽  
Department Of Health ◽  
The University

Objectives. Disease surveillance combines data collection and analysis with dissemination of findings to decision makers. The timeliness of these activities affects the ability to implement preventive measures. Influenza surveillance has traditionally been hampered by delays in both data collection and dissemination. Methods. We used statistical process control (SPC) to evaluate the daily percentage of outpatient visits with a positive point-of-care (POC) influenza test in the University of Utah Primary Care Research Network. Results. Retrospectively, POC testing generated an alert in each of 4 seasons (2004–2008, median 16 days before epidemic onset), suggesting that email notification of clinicians would be 9 days earlier than surveillance alerts posted to the Utah Department of Health website. In the 2008-09 season, the algorithm generated a real-time alert 19 days before epidemic onset. Clinicians in 4 intervention clinics received email notification of the alert within 4 days. Compared with clinicians in 6 control clinics, intervention clinicians were 40% more likely to perform rapid testing () and twice as likely to vaccinate for seasonal influenza () after notification. Conclusions. Email notification of SPC-generated alerts provided significantly earlier notification of the epidemic onset than traditional surveillance. Clinician preventive behavior was not significantly different in intervention clinics.

scholarly journals Optimization of Lyophilized LAMP and RT-PCR Reaction Mixes for Detection of Tuberculosis

2020 ◽  
Vol 4 (4) ◽  
pp. 230-236
Author(s):  
Esra Agel ◽  
Hasan Sagcan
Keyword(s):  
Rural Areas ◽  
Point Of Care ◽  
Culture Method ◽  
Molecular Techniques ◽  
Cold Chain ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Gold Standard Method ◽  
Tuberculosis Diagnosis ◽  
Peg 8000

AbstractUndoubtedly, one of the most infectious diseases in the world is tuberculosis. Key factor for tuberculosis control is to prevent possible contagion with rapid diagnosis and effective treatment. The culture method, which it takes several weeks to obtain results, is the gold standard method for laboratory diagnosis of tuberculosis. In order to prevent possible contagion of tuberculosis, diagnosis must be made in short time and treatment should be started as soon as possible. Normally, clinical samples are studied in advanced laboratories designed for this purpose. However, especially after the screening in rural areas, the transmission of the samples to the centers has many negative effects on the clinical material. Therefore, the latest trend molecular techniques in microbiological diagnosis are developing into point of care systems that can be applied in the field without laboratory infrastructure. The major challenge for molecular-based point-of-care tests is the need to store polymerase enzymes and some of the ingredients used in the cold chain. The aim of this study is to increase the resistance of the amplification reaction mixtures by lyophilizing the tuberculosis diagnosis. Lyophilization was performed on Loop-mediated isothermal amplification (LAMP) and Real-time PCR mixtures. For the lyophilization of LAMP and RT-PCR mixtures, two different experimental setups were tried from the literature except for the developed content. Chemicals such as stachyose, trehalose, glycerol and PEG 8000 are widely using as cryoprotectants. As a result, the developed content (0.5% PEG 8000, 2.0 % Stachyose) was determined the best cryoprotectant mixture. Accordingly, amplification mixtures can be produced with the developed lyophilization method and point of care kits can be developed.

scholarly journals Isothermal Amplification of Nucleic Acids Coupled with Nanotechnology and Microfluidic Platforms for Detecting Antimicrobial Drug Resistance and Beyond

2021 ◽  
Author(s):  
Seyedeh Zahra Alamolhoda ◽  
Nosratollah Zarghami ◽  
Houman Kahroba ◽  
Ahmad Mehdipour ◽  
Mohammad Pourhassan-Moghaddam ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Point Of Care ◽  
High Sensitivity ◽  
Molecular Techniques ◽  
Isothermal Amplification ◽  
High Tech ◽  
Antimicrobial Drug Resistance ◽  
Resource Limited ◽  
Accessible Point ◽  
Sensitivity Specificity

Antibiotic resistance is one of the serious health-threatening issues globally, the control of which is indispensable for rapid diagnosis and treatment because of the high prevalence and risks of pathogenicity. Traditional and molecular techniques are relatively expensive, complex, and non-portable, requiring facilities, trained personnel, and high-tech laboratories. Widespread and timely-detection is vital to the better crisis management of rapidly spreading infective diseases, especially in low-tech regions and resource-limited settings. Hence, the need for inexpensive, fast, simple, mobile, and accessible point-of-care (POC) diagnostics is highly demanding. Among different biosensing methods, the isothermal amplification of nucleic acids is favorite due to their simplicity, high sensitivity/specificity, rapidity, and portability, all because they require a constant temperature to work. Isothermal amplification methods are utilized for detecting various targets, including DNA, RNA, cells, proteins, small molecules, ions, and viruses. In this paper, we discuss various platforms, applications, and potentials of isothermal amplification techniques for biosensing of antimicrobial resistance. We also evaluate the potential of these methods, coupled with the novel and rapidly-evolving platforms offered by nanotechnology and microfluidic devices.

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