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DNA; Helicobacter pylori; drug resistance; genotype; genotyping; mutation; rapid methods
Abstract:
... Rapid detection of single-nucleotide mutations (SNMs) has played a vital role for point-of-care testing. We herein first introduced accelerated thermal cycling into conventional allele-specific qPCR (AS-qPCR), named accelerated cycling PCR (AC-PCR) to achieve rapid and sensitive detection of SNM. It could simultaneously detect 10 copies of H. pylori DNA and identify its clarithromycin-resistance g ...
Food and Drug Administration; Listeria monocytogenes; data collection; food industry; food safety; listeriosis; matrix-assisted laser desorption-ionization mass spectrometry; pathogens; statistics; support vector machines
Abstract:
... Listeria monocytogenes belongs to the category of facultative anaerobic bacteria, and is the pathogen of listeriosis, potentially lethal disease for humans. There are many similarities between L. monocytogenes and other non-pathogenic Listeria species, which causes great difficulties for their correct identification. The level of L. monocytogenes contamination in food remains high according to sta ...
... qPCR absolute quantification requires the assessment of PCR reaction efficiency. This is achieved by performing serial dilutions of gDNA. Herein, we demonstrate that when quantifying bacterial load in mixed samples, reaction efficiency should not be used as a calibration curve since gDNA isolation efficiency is neglected, significantly impacting quantification. ...
... Syphilis is a sexually transmitted disease caused by Treponema pallidum. DNA amplification methods have started to be used to facilitate diagnosis at different stages of the disease. The success of such methodologies depends on obtaining DNA from clinical samples in adequate quantity and quality for molecular reactions. There are many DNA extraction kits, but often the molecular analysis process i ...
... New techniques are needed to speed-up the identification and antimicrobial susceptibility testing (AST) of bacteria associated with bloodstream infections. Alfred 60/AST (Alifax®, Polverara, Italy) performs AST by light scattering directly from positive blood cultures. We evaluated Alfred 60/AST performances for 4 months. Each new episode of bacteraemia was included and AST were compared to either ...
... VITEK®2, MALDI-TOF MS and 16S rRNA sequencing were evaluated for the identification of aerobic endospore-forming bacteria (AEB) from a pharmaceutical facility. MALDI-TOF MS demonstrated higher accuracy compared to VITEK®2, although both databases were insufficient to identify AEB species. Sequencing was the best methodology, but unable to identify closely related species. ...
... Ozone treatment is an eco-friendly and cost-effective approach to achieve material disinfection, and this disinfection method is of utmost importance in the present global pandemic. The efficacy of ozone's oxidative potential on common microorganisms has been extensively studied, particularly in the food and water treatment industries. However, little is still understood regarding its antimicrobia ...
... The methods employed to detect non-tuberculous mycobacteria on environmental samples are essentially those classically used in clinical microbiology, which envisage a decontamination step to reduce the overgrowth of non-mycobacterial organisms before plating them on the culture medium. The aim of this study was to propose alternative culture techniques to improve non-tuberculous mycobacteria detec ...
... Blood culture, a method for identifying causative agents of bacterial sepsis, requires several days. The combination of cell-direct polymerase chain reaction and nucleic acid lateral flow immunoassay (cdPCR-NALFIA) is a simple and sensitive detection method for identifying pathogenic bacteria. Furthermore, this assay, when applied directly to blood samples yields results within 4.5 h, without requ ...
... Rapid detection methods require pre-enrichment culture in order to detect low levels of foodborne pathogens. To rapidly detect foodborne pathogens, enrichment culture processes could be replaced. Filtration and immunomagnetic separation methods have been identified to effectively concentrate and separate target pathogens from foods. In this study, a combination of filtration and immunomagnetic sep ...
... Paratuberculosis is a globally prevalent disease, that adversely affects the economy of livestock farming. Control is largely based on early detection followed by ‘Test and Cull’ or ‘Test and Segregate’ Policy. Implementation of paratuberculosis control is a special challenge due to the non-availability of point of care diagnostics (PoCD). Therefore, the present study aimed to optimize and evaluat ...
... Currently, several methods are available for the isolation of bacterial DNA and RNA. However, the diversity and complexity of cell envelope structures limit their efficiency depending on the target bacterial species. In this study, we compared the differences in yield and integrity of RNA prepared from four gram-negative and six gram-positive bacterial species using bead-beating, bacteriolytic pro ...
agar; bacterial proteins; culture media; genes; matrix-assisted laser desorption-ionization mass spectrometry; methicillin; methicillin-resistant Staphylococcus aureus; protein synthesis; yeast extract
Abstract:
... Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) is routinely used for bacterial identification in clinical laboratories. Bacterial protein expression may differ according to their growth conditions, especially the culture medium composition. We aimed to study the peak variations of Staphylococcus aureus grown on various blood agar plates (BAP), especiall ...
... Conventional diagnostic techniques using manual methods for stool examination have important limitations. Hence there is a need for improved technologies in routine clinical practice. This study aimed to compare detection rates, agreements, and diagnostic performances for stool examinations in all parameters of the complete filtration method using the Sciendox Feces Analysis System-50 automated fe ...
... Conventional microbiological methods to evaluate the in vitro antifungal activity of bioactive compounds usually consume a long time. It is also difficult to calculate different kinetic parameters. For this reason, this study aimed to evaluate the sensitivity of phytopathogenic fungi to an ethanolic extract of jackfruit leaf by the poison agar and isothermal microcalorimetry (IMC) tests. The kinet ...
... Herein we described a versatile liquid chromatographic method for detection and quantification of the total levels of two antimicrobials [i.e., streptomycin (STM) and doxycycline (DOX)], in mice plasma and selected tissues, with the aid of a single quadrupole as a detection method. The method included a few sample preparation steps, including freeze-drying and in situ triphasic solvent-assisted de ...
... A method for testing the antibiotic susceptibility of foodborne bacterial samples using a portable Raman spectrometer has been developed. Surface-enhanced Raman spectroscopy (SERS) was used to obtain the spectra from bacterial samples collected on a filter membrane after incubation with antibiotics. The SERS spectra revealed characteristic antibiotic response patterns, which can be used to assess ...
... High-throughput screening (HTS) is a present-day approach for assaying thousands of cultures in parallel. This miniaturization allows rapid screening of large number of microorganims capable of producing bio-based materials thereby meeting the demands of the ever evolving food, pharmaceutical and cosmetic industry. In this study, agar-based assays for phosphate solubilization, cellulose degradatio ...
Consolación Sánchez-Sánchez; Mercedes Aranda-Medina; Alicia Rodríguez; Alejandro Hernández; María G. Córdoba; Francisco Cuadros-Blázquez; Santiago Ruiz-Moyano
... Anaerobic digestion is a growing technology to manage organic waste and produce bioenergy. To promote this technology, it is essential to know, at the molecular level, the dynamics of microbial communities, specifically the methanogenic community. In the present study, three primer pairs were selected from seven primer pairs which were designed and tested with different concentrations and conditio ...
Bayesian theory; Brucella; algorithms; appetite; arthritis; blood; brucellosis; case studies; data collection; decision making; decision support systems; fever; hospitals; humans; livestock; pathogens; patients; therapeutics; vaccines; zoonoses
Abstract:
... Brucellosis is a common zoonotic infection of humans from livestock. This bacterial infection is acquired from infected animals and their products. The pathogen of this disease is a genus of bacilli called Brucella, and no effective vaccine has been discovered yet for the prevention of human brucellosis. The present study is mainly conducted to diagnose brucellosis accurately and timely, using Dat ...