Isothermal DNA amplification combined with lateral flow dipsticks for detection of biothreat agents.
Anal Biochem
; 560: 60-66, 2018 11 01.
Article
en En
| MEDLINE
| ID: mdl-30217500
ABSTRACT
The recently developed methods of nucleic acids isothermal amplification are promising tools for point-of-care diagnostics and in the field detection of pathogenic microorganisms. However, application of these methods outside a laboratory faces some challenges such as the rapid and sensitive detection of amplified products and the absence of cross-reactivity with genetically related microorganisms. In the presented study we compared three methods of isothermal DNA amplification loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA) and thermophilic helicase-dependent isothermal DNA amplification (tHDA), for detection of highly dangerous pathogens, such as Bacillus anthracis, Francisella tularensis and Yersinia pestis, and combined them with lateral flow dipsticks for the rapid visualization of amplified products. We observed low specificity of the three methods for B. antharcis, medium for Y. pestis and high for F. tularensis detection. Sensitivity and the detection limit were high and comparable for all the methods. We concluded that the lateral flow dipsticks have been a very useful tool for product detection of the isothermal amplification methods and enable reading the results without the use of any equipment. However, our results showed that the use of isothermal amplification methods is strongly related to the risk of false positive results.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Bacillus
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Yersinia
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Técnicas de Tipificación Bacteriana
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Técnicas de Amplificación de Ácido Nucleico
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Armas Biológicas
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Francisella tularensis
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Anal Biochem
Año:
2018
Tipo del documento:
Article