DNA Analyses in Food Safety and Quality: Current Status and Expectations

Marchelli, Rosangela; Tedeschi, Tullia; Tonelli, Alessandro

doi:10.1007/978-94-007-1226-3_2

Cited by 3 publications

(2 citation statements)

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“…DNA biosensors have gained in importance over the last few decades . Their applicability ranges from biotechnology and biomedical applications , to food safety and forensics. − Owing to their selectivity and sensitivity, DNA biosensors have become the gold standard for the diagnosis and monitoring of several types of gene-related diseases, such as cystic fibrosis and cancer …”

Section: Introductionmentioning

confidence: 99%

Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups

et al. 2018

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Biosensors and materials for biomedical applications generally require chemical functionalization to bestow their surfaces with desired properties, such as specific molecular recognition and antifouling properties. The use of modified poly(l-lysine) (PLL) polymers with appended oligo(ethylene glycol) (OEG) and thiol-reactive maleimide (Mal) moieties (PLL-OEG-Mal) offers control over the presentation of functional groups. These reactive groups can readily be conjugated to, for example, probes for DNA detection. Here we demonstrate the reliable conjugation of thiol-functionalized peptide nucleic acid (PNA) probes onto predeposited layers of PLL-OEG-Mal and the control over their surface density in the preceding synthetic step of the PLL modification with Mal groups. By monitoring the quartz crystal microbalance (QCM) frequency shifts of the binding of complementary DNA versus the density of Mal moieties grafted to the PLL, a linear relationship between probe density and PLL grafting density was found. Cyclic voltammetry experiments using Methylene Blue-functionalized DNA were performed to establish the absolute probe density values at the biosensor surfaces. These data provided a density of 1.2 × 1012 probes per cm2 per % of grafted Mal, thus confirming the validity of the density control in the synthetic PLL modification step without the need of further surface characterization.

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Section: Introductionmentioning

confidence: 99%

Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups

et al. 2018

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“…A great effort has been devoted, during the past decade, to the development of new devices for specific DNA sequences detection, due to the relevance of nucleic acid analysis in many areas, including clinical diagnostics [ 1 ], environmental monitoring and food-quality control [ 2 , 3 ]. However, diagnostic tools used to detect nucleic acids need to meet specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the whole detection assay [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Isothermal Amplification Methods for the Detection of Nucleic Acids in Microfluidic Devices

2012

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Diagnostic tools for biomolecular detection need to fulfill specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the assay. The nucleic acid amplification is a key step in DNA detection assays. It contributes to improving the assay sensitivity by enabling the detection of a limited number of target molecules. The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device. The vast majority of miniaturized systems for nucleic acid analysis exploit the polymerase chain reaction (PCR) amplification method, which requires repeated cycles of three or two temperature-dependent steps during the amplification of the nucleic acid target sequence. In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features. Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.

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