Rapid Listeria Monocytogenes Assay Based On Helicase Dependent Amplification (Hda) And Nucleic Acid Hybridization In Blue Silver Nanoplates
| Autor: | Kankanit Pisamayarom, Piyasak Chaumpluk |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Detection limit Chemistry 030106 microbiology 010402 general chemistry medicine.disease_cause 01 natural sciences Listeria Monocytogenes Helicase Dependent Amplification (HDA) Blue Silver Nanoplates (Agnpls) Frozen Seafood 0104 chemical sciences 03 medical and health sciences Nucleic acid thermodynamics chemistry.chemical_compound Listeria monocytogenes Biochemistry Rapid assay medicine Oligomer restriction Helicase-dependent amplification Gene DNA |
| DOI: | 10.5281/zenodo.1043426 |
| Popis: | Listeria monocytogenes, a foodborne pathogen, is considered as one of the major problems in food safety. With strong safety regulations, a monitoring measure is essential for protecting the health and safety of consumers. Thus, a reliable monitoring method is required. In this study, a rapid assay based on a combination of helicase dependent amplification (HDA) and DNA signal detection via nucleic acid hybridization in blue silver nanoplates (AgNPls) was established. The assay started directly after short term enrichment in terrific broth using cotton ball swapping technique on seafood surface. A HDA amplification of hly gene of L. monocytogenes at 65 °C allowed DNA signals to be increased, whereas the rendered DNA products were detected via nucleic acid hybridization with an oligonucleotide probe in AgNPls solution. The positive specimens induced blue silver nanoplates’ aggregation resulting in pale gray change to colorless, while the negative specimens showed the blue color of non-aggregated nanoplates. The method had a detection limit at 100 copies of L. monocytogenes DNA per 50 g of sample. This method was rapid, simple, did not require laboratory facilities and was suitable for field food safety monitoring |
| Databáze: | OpenAIRE |
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