A novel visual-mixed-dye for LAMP and its application in the detection of foodborne pathogens.
Anal Biochem
; 574: 1-6, 2019 06 01.
Article
en En
| MEDLINE
| ID: mdl-30862446
ABSTRACT
Loop-mediated isothermal amplification (LAMP) has been widely applied for the detection of foodborne pathogens. Obtaining a simple, accurate readout of LAMP reaction results is crucial. Herein, a visual-mixed-dye (VMD) containing calcein (precombined with MnCl2) and hydroxynaphthol blue (HNB) for LAMP end-point detection was developed. The optimal final concentrations of these components in VMD were 25 and 300⯵M, respectively. Due to the formation of HNBMn2+ during the LAMP reaction, the VMD-loaded assay exhibited superior visual properties, changing from light gray (negative) to dark blue (positive) under natural light. Additionally, compared with traditional single calcein or HNB dye, a weakly positive result with the VMD was purple, making it easier to distinguish by the naked eye. The visual sensitivity reached down to 20.9 copies/µL, which was comparable to that based on fluorescence. In food-contaminated samples, the practicality of VMD was verified by Vibrio parahaemolyticus and Staphylococcus aureus detection with excellent specificity. Moreover, the VMD was stable over a period of 3 months. Collectively, these findings establish the VMD as a novel dye for LAMP.
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Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Técnicas de Amplificación de Ácido Nucleico
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Colorantes
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Microbiología de Alimentos
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Bacterias Gramnegativas
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Bacterias Grampositivas
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Año:
2019
Tipo del documento:
Article