presents one of the main waterborne public health threats due to its resistance to chlorine disinfection and ability to cause large-scale outbreaks. The standard method used in the UK water industry for detection and enumeration of is based on fluorescence microscopy and is laborious and expensive. Molecular methods such as quantitative polymerase chain reaction (qPCR) can be more amenable to streamlining through automation, improving workflows and standardizing procedures. The null hypothesis was that there was no difference in the detection or enumeration between the standard method and a qPCR. We aimed to develop and evaluate a qPCR for the detection and enumeration of in drinking water, and to compare the assay with the standard method used in the UK. We first developed and evaluated a qPCR method by incorporating an internal amplification control and calibration curve into a real-time PCR currently used for genotyping. Then we compared the qPCR assay with the standard method of immunofluorescent microscopy for the detection and enumeration of 10 and 100 oocysts in 10 l of artificially contaminated drinking water. The results demonstrated that detection of by this qPCR was reliable at low numbers of oocysts; however, enumeration was less reliable and more variable than immunofluorescence microscopy. Despite these results, qPCR offers practical advantages over microscopy. There is potential for the use of PCR-based methods for analysis if parts of the upstream sample preparation are revised, and alternative technologies for enumeration (such as digital PCR) are also explored to improve analytical sensitivity.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1099/jmm.0.001715 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Detection rate for ESR1 mutations is higher in circulating-tumor-cell-derived genomic DNA than in paired plasma cell-free DNA samples as revealed by ddPCR.
Mol Oncol
January 2025
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Greece.
Plasma cell-free DNA (cfDNA) analysis to track estrogen receptor 1 (ESR1) mutations is highly beneficial for the identification of tumor molecular dynamics and the improvement of personalized treatments for patients with metastatic breast cancer (MBC). Plasma-cfDNA is, up to now, the most frequent liquid biopsy analyte used to evaluate ESR1 mutational status. Circulating tumor cell (CTC) enumeration and molecular characterization analysis provides important clinical information in patients with MBC.
View Article and Find Full Text PDFEarly detection of bacterial pneumonia by characteristic induced odor signatures.
BMC Infect Dis
December 2024
Department of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, University of Zürich, Zurich, 8097, Switzerland.
Introduction: The ability to detect pathogenic bacteria before the onsets of severe respiratory symptoms and to differentiate bacterial infection allows to improve patient-tailored treatment leading to a significant reduction in illness severity, comorbidity as well as antibiotic resistance. As such, this study refines the application of the non-invasive Secondary Electrospray Ionization-High Resolution Mass Spectrometry (SESI-HRMS) methodology for real-time and early detection of human respiratory bacterial pathogens in the respiratory tract of a mouse infection model.
Methods: A real-time analysis of changes in volatile metabolites excreted by mice undergoing a lung infection by Staphylococcus aureus or Streptococcus pneumoniae were evaluated using a SESI-HRMS instrument.
Detection of Putative Ligand Dissociation Pathways in Proteins Using Site-Identification by Ligand Competitive Saturation.
J Chem Inf Model
December 2024
Computer-Aided Drug Design Center, Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland Baltimore, Baltimore, Maryland 21201, United States.
Drug efficacy often correlates better with dissociation kinetics than binding affinity alone. To study binding kinetics computationally, it is necessary to identify all of the possible ligand dissociation pathways. The site identification by ligand competitive saturation (SILCS) method involves the precomputation of a set of maps (FragMaps), which describe the free energy landscapes of typical chemical functionalities in and around a target protein or RNA.
View Article and Find Full Text PDFAn Image Processing Tool for Automated Quantification of Bacterial Burdens in Zebrafish Larvae.
Zebrafish
December 2024
Department of Medicine, Molecular Immunity Unit, Cambridge Institute of Therapeutic Immunology and Infectious Diseases, University of Cambridge, Cambridge, UK.
Zebrafish larvae are used to model the pathogenesis of multiple bacteria. This transparent model offers the unique advantage of allowing quantification of fluorescent bacterial burdens (fluorescent pixel counts [FPC]) by facile microscopical methods, replacing enumeration of bacteria using time-intensive plating of lysates on bacteriological media. Accurate FPC measurements require laborious manual image processing to mark the outside borders of the animals so as to delineate the bacteria inside the animals from those in the culture medium that they are in.
View Article and Find Full Text PDFAn integrated microflow cytometry platform with artificial intelligence capabilities for point-of-care cellular phenotype analysis.
Biosens Bioelectron
March 2025
Department of Biotechnology, National Formosa University, No. 64, Wunhua Rd, Huwei Township, Yunlin County, 63201, Taiwan. Electronic address:
The EZ DEVICE is an integrated fluorescence microflow cytometer designed for automated cell phenotyping and enumeration using artificial intelligence (AI). The platform consists of a laser diode, optical filter, objective lens, CMOS image sensor, and microfluidic chip, enabling automated sample pretreatment, labeling, and detection within a single compact unit. AI algorithms segment and identify objects in images captured by the CMOS sensor at 532 and 586 nm emission wavelengths.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!