Multiplex loop-mediated isothermal amplification-based lateral flow dipstick for simultaneous detection of 3 food-borne pathogens in powdered infant formula
| Autor: | Xinyan Yang, Zhao Yueming, Yujun Jiang, John L. McKillip, Shiqian Fu, Edward M. Fox, Sihan Chen, Chaoxin Man |
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| Rok vydání: | 2019 |
| Předmět: |
Streptavidin
Salmonella Staphylococcus aureus Loop-mediated isothermal amplification Metal Nanoparticles medicine.disease_cause Sensitivity and Specificity 03 medical and health sciences chemistry.chemical_compound Limit of Detection Genetics medicine Multiplex Cronobacter 030304 developmental biology DNA Primers 0303 health sciences Chromatography biology 0402 animal and dairy science food and beverages 04 agricultural and veterinary sciences Dipstick Amplicon biology.organism_classification 040201 dairy & animal science Infant Formula chemistry Animal Science and Zoology Gold Powders Nucleic Acid Amplification Techniques Food Science |
| Zdroj: | Journal of dairy science. 103(5) |
| ISSN: | 1525-3198 |
| Popis: | In this study, we established a rapid, simple, and sensitive method for visual and point-of-care detection of Salmonella spp., Cronobacter spp., and Staphylococcus aureus in powdered infant formula (PIF) based on multiplex loop-mediated isothermal amplification (mLAMP) combined with lateral flow dipstick (LFD). Three different species-specific target genes, siiA of Salmonella spp., internal transcribed space (ITS) of Cronobacter spp., and nuc of Staph. aureus, were applied in the mLAMP with biotin-, digoxin-, and Texas Red-modified forward inner primers and fluorescein isothiocyanate (FITC)-modified backward inner primers. After mLAMP, a large number of modified amplicons were detected with LFD; one end of the amplicon was conjugated to the anti-FITC antibody on gold nanoparticles and the other end to streptavidin (anti-digoxin or anti-Texas Red antibody) on test lines. Visual inspection of the device relies on the presence of a red band formed by accumulation of sandwich composites. The detection limits of this mLAMP-LFD assay for Salmonella spp., Cronobacter spp., and Staph. aureus in PIF without enrichment were 4.2, 2.6, and 3.4 cfu/g, respectively. The whole method can be completed in less than 1 h. Thus, mLAMP-LFD is a rapid and efficient method for simultaneously detecting Salmonella spp., Cronobacter spp., and Staph. aureus in PIF. |
| Databáze: | OpenAIRE |
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