Rapid and Highly Sensitive Detection by a Real-time Polymerase Chain Reaction Using a Chip Coated with Its Reagents

Furutani, Shunsuke; Naruishi, Nahoko; Saito, Masato; Tamiya, Eiichi; Fuchiwaki, Masaki; Nagai, Hidenori

doi:10.2116/analsci.30.569

Cited by 9 publications

(4 citation statements)

References 21 publications

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“…However, depending on the type of sample, adjustments during sample preparation may be required. Furutani et al ( 2014 ) reduced the run time to 8 min for the detection of E. coli with a high sensitivity of 4 cells μL −1 using a microfluidic RT-PCR chip coated with PCR reagents. The multiplexed detection of waterborne pathogens on a microfluidic RT-PCR chip has already been achieved (Ramalingam et al 2010 ).…”

Section: Future Trendsmentioning

confidence: 99%

Trends and advances in food analysis by real-time polymerase chain reaction

et al. 2016

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Analyses to ensure food safety and quality are more relevant now because of rapid changes in the quantity, diversity and mobility of food. Food-contamination must be determined to maintain health and up-hold laws, as well as for ethical and cultural concerns. Real-time polymerase chain reaction (RT-PCR), a rapid and inexpensive quantitative method to detect the presence of targeted DNA-segments in samples, helps in determining both accidental and intentional adulterations of foods by biological contaminants. This review presents recent developments in theory, techniques, and applications of RT-PCR in food analyses, RT-PCR addresses the limitations of traditional food analyses in terms of sensitivity, range of analytes, multiplexing ability, cost, time, and pointof-care applications. A range of targets, including species of plants or animals which are used as food ingredients, food-borne bacteria or viruses, genetically modified organisms, and allergens, even in highly processed foods can be identified by RT-PCR, even at very low concentrations. Microfluidic RT-PCR eliminates the separate sample-processing step to create opportunities for point-of-care analyses. We also cover the challenges related to using RT-PCR for food analyses, such as the need to further improve sample handling.

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Section: Future Trendsmentioning

confidence: 99%

Trends and advances in food analysis by real-time polymerase chain reaction

et al. 2016

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“…Rapid PCR conditions were optimized by adding a fluorescent dye to the PCR reagent in GeneSoC. In our previous studies, SpeedSTAR DNA HS Polymerase was used as the optimal enzyme for rapid fluorescence-based PCR amplification using the TaqMan probe method [ 15 , 17 , 18 ]. In the present study, we additionally confirmed rapid PCR amplification using KOD One polymerase, which lacks 5′ nuclease activity.…”

Section: Resultsmentioning

confidence: 99%

Rapid DNA Sequencing Technology Based on the Sanger Method for Bacterial Identification

Furutani

Kawai

et al. 2022

Sensors

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Antimicrobial resistance, a global health concern, has been increasing due to inappropriate use of antibacterial agents. To facilitate early treatment of sepsis, rapid bacterial identification is imperative to determine appropriate antibacterial agent for better therapeutic outcomes. In this study, we developed a rapid PCR method, rapid cycle sequencing, and microchip electrophoresis, which are the three elemental technologies for DNA sequencing based on the Sanger sequencing method, for bacterial identification. We achieved PCR amplification within 13 min and cycle sequencing within 14 min using a rapid thermal cycle system applying microfluidic technology. Furthermore, DNA analysis was completed in 14 min by constructing an algorithm for analyzing and performing microchip electrophoresis. Thus, the three elemental Sanger-based DNA sequencing steps were accomplished within 41 min. Development of a rapid purification process subsequent to PCR and cycle sequence using a microchip would help realize the identification of causative bacterial agents within one hour, and facilitate early treatment of sepsis.

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“…Use of TaqMan® probe DNA is a powerful tool for analyzing sequences of PCR products. However, real-time PCR systems using the TaqMan® probe can be affected by sample impurities, and the inhibitory activity of impurities against DNA polymerase depends on the type of DNA polymerase [44][45][46][47][48]. This observation suggests that suitable DNA polymerases may not always be used in real-time PCR because the system requires DNA polymerases that exhibit exonuclease activity.…”

Section: Comparison With Real-time Pcr Technologymentioning

confidence: 99%

DNA Detection Technology Using Zinc Finger Protein

Abe¹,

Ikebukuro²

2015

J Microb Biochem Technol

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With recent advances in nanotechnology and sequencing technology, DNA diagnostic technology is becoming practical and is advancing at a fast pace. In these detection systems, mainly PCR (particularly real-time PCR) and DNA probe hybridization techniques are used. We suggest that detecting PCR products using double-stranded DNA (dsDNA) is more convenient and powerful compared with DNA probe hybridization technique. Zinc finger protein is the major DNA binding protein in nature and it recognizes dsDNA with sequence specific manner. Additionally, by changing its amino acids sequence, we can design it to recognize the desired DNA sequence to some extent. Using zinc finger protein for DNA detection element, simple, accurate and sensitive DNA detection can be achieved. In this review, dsDNA detection using zinc finger protein is described and compared with recent advanced technology.

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Rapid and Highly Sensitive Detection by a Real-time Polymerase Chain Reaction Using a Chip Coated with Its Reagents

Cited by 9 publications

References 21 publications

Trends and advances in food analysis by real-time polymerase chain reaction

Trends and advances in food analysis by real-time polymerase chain reaction

Rapid DNA Sequencing Technology Based on the Sanger Method for Bacterial Identification

DNA Detection Technology Using Zinc Finger Protein

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