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Loop-mediated Isothermal Amplification LAMP Assays for the Species-specific Detection of Eimeria that Infect Chickens
Published on: February 20, 2015
Direct loop-mediated isothermal amplification assay for on-site detection of Staphylococcus aureus
Xiaolan Tian1, Junli Feng1,2, Yi Wang1
1Institute of Seafood, Zhejiang Gongshang University, Hangzhou 310012, PR China.
Abstract:
Staphylococcus aureus (S. aureus) is a major human pathogen that may produce a variety of toxins and cause staphylococcal food poisoning. In the present study, a direct loop-mediated isothermal amplification (LAMP) assay was developed and validated to detect S. aureus in food samples. Without prior cultural enrichment and DNA extraction steps, bacterial DNA was released by heating at 100°C for 5 min and directly subjected to LAMP assay. Using a set of LAMP primers recognizing six distinct regions of nuc gene, the developed direct LAMP assay was highly specific, and the analytical sensitivity was determined to be 7.6 × 102 CFU/mL. Moreover, with the pre-mixed LAMP reagents stored at -20°C, the entire assay should be finished within 40 min. These features greatly simplified the operating procedure and made the direct LAMP a powerful tool for rapid and on-site detection of S. aureus in food samples.
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