Objectives: and are three toxin producing bacteria over the world, especially in Iran, and it is essential to find a certain, rapid procedure to identify these microorganisms. In this research, these bacteria were simultaneously detected by multiplex PCR technique in foods.
Materials And Methods: The primary approval of bacterial strains was performed by biochemical tests. PCR primers were designed based on the nucleotide sequences of the gene of . , the gene of and the C gene of . The specificity of Multiplex PCR method was determined using seven food poisoning bacteria including . To confirm the reaction, DNA extraction was performed from 30 food samples (milk), and gene amplification was performed by PCR. The length of amplified fragments was 300 bp, 210 bp and 160 bp for , and genes, respectively.
Results: The detection limits of the PCR method were 5, 4 and 3 pg for , and , respectively. Specifisity test showed that this reaction is spesific to these 3 bacteria.
Conclusion: In this study, we introduced a new multiplex PCR method for simultsnus detection of and . These results can be used for detection of other toxin producing bacteria in food.
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http://dx.doi.org/10.22038/IJBMS.2017.9275 | DOI Listing |
Toxicol Pathol
January 2025
Johnson & Johnson Innovative Medicine, La Jolla, California, USA.
Small interfering RNAs (siRNAs) have been successfully used as therapeutics to silence disease-causing genes when conjugated to ligands or formulated in lipid nanoparticles to target relevant cell types for efficacy while sparing other cells for safety. To support the development of new methods for delivery of siRNA therapeutics, we developed and characterized a panel of antibodies generated against chemically modified nucleotides used in therapeutic siRNA molecules, identifying a monoclonal antibody that detects a broad range of siRNA representing distinct sequences and modification patterns. By integrating this anti-siRNA antibody with additional reagents, we created a multiplex siRNA immunoassay that simultaneously quantifies siRNA uptake, trafficking, and silencing activity.
View Article and Find Full Text PDFJ Infect Dis
January 2025
Department of Pulmonary and Critical Care Medicine, Taihe Hospital, Hubei University of Medicine, Shiyan, 442000, Hubei, P.R. China.
Background And Objective: Multiplex polymerase chain reaction (PCR)-based targeted next-generation sequencing (tNGS) is a promising tool for distinguishing lower respiratory tract infections (LRTIs) in clinical practice, and its detectable pathogen spectrum can cover more than 95% of clinical cases. but there is limited information on systematic evaluation of the clinical use of multiplex PCR-based tNGS (mp-tNGS) in IPA cases. We aim to assess mp-tNGS in bronchoalveolar lavage fluid (BALF) for Aspergillus detection in suspected IPA patients, and to provide a reliable basis for initiating antifungal therapy without microbiological or histopathological evidence.
View Article and Find Full Text PDFJ Virol Methods
January 2025
Centrillion Technologies, Palo Alto, CA 94303.
Humanity faces an ongoing battle at the microscopic level to identify, contain, and treat outbreaks of numerous pathogens each year. Global genomic surveillance is the first step in monitoring outbreaks, but high-throughput methods are expensive and time-consuming. To solve this problem, we designed and manufactured a resequencing microarray capable of identifying 35 viral pathogens, 21 pathogenic bacteria, 16 antibiotic resistance genes, and 6 controls.
View Article and Find Full Text PDFBMC Microbiol
January 2025
Cellular Interactions of Bacterial Pathogens, Centre for Biological Threats and Special Pathogens, Highly Pathogenic Microorganisms (ZBS 2), Robert Koch Institute, Seestraße 10, 13353, Berlin, Germany.
Background: The zoonotic and highly infectious pathogen Francisella tularensis is the etiological agent of tularemia. Tularemia in humans is mainly caused by F. tularensis subspecies tularensis and holarctica, but Francisella species like F.
View Article and Find Full Text PDFInfection
January 2025
Institute of Population Health Sciences, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County, 35053, Taiwan.
Purpose: Rapid detection of drug resistance in Mycobacterium tuberculosis (Mtb) from clinical samples facilitates the timely provision of optimal treatment regimens for tuberculosis (TB) patients.
Methods: In November, 2023, the WHO released its second catalogue of resistance-conferring mutations in Mtb. Utilizing this information, we developed a single 17-plex PCR assay covering 16 key resistance genes and modified thermo-protection buffer to amplify 30 kbp DNA directly from sputum samples for nanopore sequencing.
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