Simultaneous Detection of Escherichia coli and Agrobacterium tumefaciens by Using Gold Nanoparticle Enhanced Polymerase Chain Reaction

Micro, Journal Year: 2025, Volume and Issue: 5(1), P. 9 - 9

Published: Feb. 28, 2025

Escherichia coli (E. coli) and Agrobacterium tumefaciens (A. tumefaciens) are bacterial species commonly found in the environment, they can do much harm to humans, animals plants. As a result, it is necessary find an accurate, rapid, simple method detect concentrations of them, polymerase chain reaction (PCR) one most suitable candidates. In this study, gold nanoparticles (GNPs) enhanced was developed, simultaneously target specific genes, 16S rDNA E. Tms1 A. tumefaciens. PCR amplification times (CT values) were seen be lowered significantly by incorporation GNPs. The fluorescence intensities quantitative amplifications both reached maximum after around 40 cycles, yield (maximum intensity) proportional absorbance at 495 nm corresponding UV-vis spectra. GNPs enhance tumefaciens, smaller sized (average 13 nm) showed better enhancement effect compared larger 30 nm). Conventional that could detected together with limit detection 10 CFU/mL for each bacterium, using nm. results study lead improvement multiplex different bacteria simultaneously.

Language: Английский

Rapid Universal Detection of High‐Risk and Low‐Abundance Microbial Contaminations in CAR‐T Cell Therapy

Small Methods, Journal Year: 2025, Volume and Issue: unknown

Published: March 30, 2025

Abstract Live microbial contamination poses high risks to cell and gene therapies, threatening manufacturing processes patient safety. Rapid, sensitive detection of live microbes in complex environments, such as CAR‐T cultures, remains an urgent need. Here, innovative sample‐to‐result workflow is introduced using digital loop‐mediated isothermal amplification (dLAMP), enhanced by Electrostatic Microfiltration (EM)‐based enrichment, for rapid sterility testing. By rationally designing primers targeting 16S 18S rRNA, dLAMP assay enables both universal (covering >80% known species) strain‐specific identification bacterial fungal contaminants spent medium final products, directly from microorganism lysates. Enhanced EM‐based enrichment low‐abundance microbes, the achieves unparalleled sensitivity speed, detecting levels low 1 CFU/mL cultures within 6 h. Compared qPCR 14‐day compendial methods, approach demonstrates superior accuracy significantly faster turnaround times. This holds transformative potential real‐time monitoring therapy safety assessments products prior infusion. Beyond therapy, method broadly applicable infectious disease diagnostics, biomanufacturing monitoring, food safety, environmental surveillance.

Language: Английский

Progress in Separation and Detection of Foodborne Bacteria for Food Safety

Current Opinion in Food Science, Journal Year: 2024, Volume and Issue: unknown, P. 101266 - 101266

Published: Dec. 1, 2024

Language: Английский