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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Santiago-Felipe, S. Tortajada-Genaro, L. A. Puchades, R. Maquieira, A. |
| Description | Country affiliation: Spain Author Affiliation: Santiago-Felipe S ( Centro de Reconocimiento Molecular y Desarrollo Tecnológico (IDM)-Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.); Tortajada-Genaro LA ( Centro de Reconocimiento Molecular y Desarrollo Tecnológico (IDM)-Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.); Puchades R ( Centro de Reconocimiento Molecular y Desarrollo Tecnológico (IDM)-Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.); Maquieira A ( Centro de Reconocimiento Molecular y Desarrollo Tecnológico (IDM)-Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain. Electronic address: amaquieira@qim.upv.es.) |
| Abstract | Polymerase chain reaction in conjunction with enzyme-linked immunosorbent assay (PCR-ELISA) is a well-established technique that provides a suitable rapid, sensitive, and selective method for a broad range of applications. However, the need for precise rapid temperature cycling of PCR is an important drawback that can be overcome by employing isothermal amplification reactions such as recombinase polymerase amplification (RPA). The RPA-ELISA combination is proposed for amplification at a low, constant temperature (40°C) in a short time (40 min), for the hybridisation of labelled products to specific 5'-biotinylated probes/streptavidin in coated microtiter plates at room temperature, and for detection by colorimetric immunoassay. RPA-ELISA was applied to screen common safety threats in foodstuffs, such as allergens (hazelnut, peanut, soybean, tomato, and maize), genetically modified organisms (P35S and TNOS), pathogenic bacteria (Salmonella sp. and Cronobacter sp.), and fungi (Fusarium sp.). Satisfactory sensitivity and reproducibility results were achieved for all the targets. The RPA-ELISA technique does away with thermocycling and provides a suitable sensitive, specific, and cost-effective method for routine applications, and proves particularly useful for resource-limited settings. |
| File Format | HTM / HTML |
| ISSN | 00032670 |
| Volume Number | 811 |
| e-ISSN | 18734324 |
| Journal | Analytica Chimica Acta |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2014-02-06 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Discipline Chemistry Dna-directed Dna Polymerase Metabolism Dna Analysis Enzyme-linked Immunosorbent Assay Food Analysis Methods Nucleic Acid Amplification Techniques Recombinases Allergens Cronobacter Genetics Dna, Bacterial Dna, Fungal Dna, Plant Food Microbiology Fusarium Plants Plants, Genetically Modified Salmonella Temperature Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry |
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