Magneto immunoseparation of pathogenic bacteria and electrochemical magneto genosensing of the double-tagged amplicon
| Autor: | Salvador Alegret, Jordi Barbé, María Isabel Pividori, Anabel Lermo, Susana Liébana, Susana Campoy |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
DNA
Bacterial Salmonella Materials science food.ingredient Hot Temperature Time Factors Bioengineering Biosensing Techniques medicine.disease_cause Polymerase Chain Reaction Microbiology Analytical Chemistry Magnetics food Culture Techniques Skimmed milk medicine Electrochemistry Food microbiology Animals Humans Molecular Biology Magneto Escherichia coli Immunoassay Chromatography biology Chemistry Salmonella enterica Pathogenic bacteria General Medicine Amplicon biology.organism_classification Enterobacteriaceae Milk Biochemistry Food Microbiology Microscopy Electron Scanning Bacteria Genome Bacterial Biotechnology |
| Zdroj: | Analytical chemistry. 81(14) |
| ISSN: | 1520-6882 |
| Popis: | A rapid and sensitive method for the detection of food pathogenic bacteria is reported. In this approach, the bacteria are captured and preconcentrated from food samples with magnetic beads by immunological reaction with the specific antibody against Salmonella. After the lysis of the captured bacteria, further amplification of the genetic material by PCR with a double-tagging set of primers is performed to confirm the identity of the bacteria. Both steps are rapid alternatives to the time-consuming classical selective enrichment and biochemical/serological tests. The double-tagged amplicon is then detected by electrochemical magneto genosensing. The "IMS/double-tagging PCR/m-GEC electrochemical genosensing" approach is used for the first time for the sensitive detection of Salmonella artificially inoculated into skim milk samples. A limit of detection of 1 CFU mL(-1) was obtained in 3.5 h without any pretreatment, in LB broth and in milk diluted 1/10 in LB. If the skim milk is pre-enriched for 6 h, the method is able to feasibly detect as low as 0.04 CFU mL(-1) (1 CFU in 25 g of milk) with a signal-to-background ratio of 20. Moreover, the method is able to clearly distinguish between pathogenic bacteria such as Salmonella and Escherichia coli. The features of this approach are discussed and compared with classical culture methods and PCR-based assay. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|