Simultaneous Detection of <i>Escherichia coli</i> and <i>Agrobacterium tumefaciens</i> by Using Gold Nanoparticle Enhanced Polymerase Chain Reaction
<i>Escherichia coli</i> (<i>E. coli</i>) and <i>Agrobacterium tumefaciens</i> (<i>A. tumefaciens</i>) are bacterial species commonly found in the environment, and they can do much harm to humans, animals and plants. As a result, it is necessary to find...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-02-01
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| Series: | Micro |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2673-8023/5/1/9 |
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| Summary: | <i>Escherichia coli</i> (<i>E. coli</i>) and <i>Agrobacterium tumefaciens</i> (<i>A. tumefaciens</i>) are bacterial species commonly found in the environment, and they can do much harm to humans, animals and plants. As a result, it is necessary to find an accurate, rapid, simple method to detect the concentrations of them, and polymerase chain reaction (PCR) is one of the most suitable candidates. In this study, a gold nanoparticles (GNPs) enhanced polymerase chain reaction was developed, to simultaneously target the specific genes, 16S rDNA of <i>E. coli</i> and Tms1 of <i>A. tumefaciens</i>. PCR amplification times (CT values) of <i>E. coli</i> and <i>A. tumefaciens</i> were seen to be lowered significantly by the incorporation of GNPs. The fluorescence intensities in quantitative PCR amplifications of both <i>E. coli</i> and <i>A. tumefaciens</i> reached the maximum after around 40 cycles, and the PCR yield (maximum fluorescence intensity) was proportional to the maximum absorbance at 495 nm in the corresponding UV-vis spectra. GNPs were found to enhance the PCR yield of both <i>E. coli</i> and <i>A. tumefaciens</i>, and smaller sized GNPs (average 13 nm) showed a better enhancement effect compared to larger sized GNPs (average 30 nm). Conventional PCR showed that both <i>E. coli</i> and <i>A. tumefaciens</i> could be detected together with limit of detection of 10 CFU/mL for each bacterium, using GNPs of 13 nm. The results of this study could lead to improvement of multiplex PCR that can detect different bacteria species simultaneously. |
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| ISSN: | 2673-8023 |