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One nanoprobe, two pathogens: gold nanoprobes multiplexing for point-of-care.

Veigas B, Pedrosa P, Carlos FF, Mancio-Silva L, Grosso AR, Fortunato E, Mota MM, Baptista PV - J Nanobiotechnology (2015)

Bottom Line: Following characterisation, the developed gold-nanoprobe allowed detection of either target in individual samples or in samples containing both DNA species with the same efficacy.Using one single probe via the non-cross-linking colorimetric methodology it is possible to identify multiple targets in one sample in one reaction.This proof-of-concept approach may easily be integrated into sensing platforms allowing for fast and simple multiplexing of Au-nanoprobe based detection at point-of-need.

View Article: PubMed Central - PubMed

Affiliation: Nanomedicine@FCT, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, CIGMH, UCIBIO, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal. bmrveigas@gmail.com.

ABSTRACT

Background: Gold nanoparticles have been widely employed for biosensing purposes with remarkable efficacy for DNA detection. Amongst the proposed systems, colorimetric strategies based on the remarkable optical properties have provided for simple yet effective sequence discrimination with potential for molecular diagnostics at point of need. These systems may also been used for parallel detection of several targets to provide additional information on diagnostics of pathogens.

Results: For the first time, we demonstrate that a single Au-nanoprobe may provide for detection of two distinct targets (pathogens) allowing colorimetric multi-target detection. We demonstrate this concept by using one single gold-nanoprobe capable to detect members of the Mycobacterium tuberculosis complex and Plasmodium sp., the etiologic agents of tuberculosis and malaria, respectively. Following characterisation, the developed gold-nanoprobe allowed detection of either target in individual samples or in samples containing both DNA species with the same efficacy.

Conclusions: Using one single probe via the non-cross-linking colorimetric methodology it is possible to identify multiple targets in one sample in one reaction. This proof-of-concept approach may easily be integrated into sensing platforms allowing for fast and simple multiplexing of Au-nanoprobe based detection at point-of-need.

No MeSH data available.


Related in: MedlinePlus

Multiplexing Au-nanoprobe based detection. DNA extracted from several biological fluids is amplified (e.g. PCR) and then a single Au-nanoprobe is used to detect presence of pathogens. Depicted is the detection of M. tuberculosis and P. falciparum. Map shows the geografical differences of Malaria and TB incidence where the highest rates are found predominantly in low-income countries in Africa and South America. This approach can be transposed into a simple and portable molecular diagnostic platform to be used at peripheral laboratories and/or point-of-need, coupled to a “smartphone” for data analysis and location metadata for real-time epidemiologic data [15].
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Fig1: Multiplexing Au-nanoprobe based detection. DNA extracted from several biological fluids is amplified (e.g. PCR) and then a single Au-nanoprobe is used to detect presence of pathogens. Depicted is the detection of M. tuberculosis and P. falciparum. Map shows the geografical differences of Malaria and TB incidence where the highest rates are found predominantly in low-income countries in Africa and South America. This approach can be transposed into a simple and portable molecular diagnostic platform to be used at peripheral laboratories and/or point-of-need, coupled to a “smartphone” for data analysis and location metadata for real-time epidemiologic data [15].

Mentions: Efforts have also been made to extend these system to allow for extra layers of information in a single reaction, either by exploring the differential kinetics of aggregation and hybridization efficiencies or relying on the use of additional plasmon signatures of NPs (e.g. gold:silver alloy nanoprobes) [18–21]. Here, we demonstrate that the Au-nanoprobe approach may be easily extended towards the simultaneous detection of two pathogens of interest with a single multi-sequence functionalized Au-nanoprobe to evaluate in a single test the presence/absence of any of the pathogens. To enhance the detection potential of this approach for MTBC and Plasmodium sp., we performed a preparative multiplex amplification of the respective specific loci—see Fig. 1. This concept extends the colorimetric detection to multiplexing suitable for point-of-need molecular screening.Fig. 1


One nanoprobe, two pathogens: gold nanoprobes multiplexing for point-of-care.

Veigas B, Pedrosa P, Carlos FF, Mancio-Silva L, Grosso AR, Fortunato E, Mota MM, Baptista PV - J Nanobiotechnology (2015)

Multiplexing Au-nanoprobe based detection. DNA extracted from several biological fluids is amplified (e.g. PCR) and then a single Au-nanoprobe is used to detect presence of pathogens. Depicted is the detection of M. tuberculosis and P. falciparum. Map shows the geografical differences of Malaria and TB incidence where the highest rates are found predominantly in low-income countries in Africa and South America. This approach can be transposed into a simple and portable molecular diagnostic platform to be used at peripheral laboratories and/or point-of-need, coupled to a “smartphone” for data analysis and location metadata for real-time epidemiologic data [15].
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4527100&req=5

Fig1: Multiplexing Au-nanoprobe based detection. DNA extracted from several biological fluids is amplified (e.g. PCR) and then a single Au-nanoprobe is used to detect presence of pathogens. Depicted is the detection of M. tuberculosis and P. falciparum. Map shows the geografical differences of Malaria and TB incidence where the highest rates are found predominantly in low-income countries in Africa and South America. This approach can be transposed into a simple and portable molecular diagnostic platform to be used at peripheral laboratories and/or point-of-need, coupled to a “smartphone” for data analysis and location metadata for real-time epidemiologic data [15].
Mentions: Efforts have also been made to extend these system to allow for extra layers of information in a single reaction, either by exploring the differential kinetics of aggregation and hybridization efficiencies or relying on the use of additional plasmon signatures of NPs (e.g. gold:silver alloy nanoprobes) [18–21]. Here, we demonstrate that the Au-nanoprobe approach may be easily extended towards the simultaneous detection of two pathogens of interest with a single multi-sequence functionalized Au-nanoprobe to evaluate in a single test the presence/absence of any of the pathogens. To enhance the detection potential of this approach for MTBC and Plasmodium sp., we performed a preparative multiplex amplification of the respective specific loci—see Fig. 1. This concept extends the colorimetric detection to multiplexing suitable for point-of-need molecular screening.Fig. 1

Bottom Line: Following characterisation, the developed gold-nanoprobe allowed detection of either target in individual samples or in samples containing both DNA species with the same efficacy.Using one single probe via the non-cross-linking colorimetric methodology it is possible to identify multiple targets in one sample in one reaction.This proof-of-concept approach may easily be integrated into sensing platforms allowing for fast and simple multiplexing of Au-nanoprobe based detection at point-of-need.

View Article: PubMed Central - PubMed

Affiliation: Nanomedicine@FCT, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, CIGMH, UCIBIO, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal. bmrveigas@gmail.com.

ABSTRACT

Background: Gold nanoparticles have been widely employed for biosensing purposes with remarkable efficacy for DNA detection. Amongst the proposed systems, colorimetric strategies based on the remarkable optical properties have provided for simple yet effective sequence discrimination with potential for molecular diagnostics at point of need. These systems may also been used for parallel detection of several targets to provide additional information on diagnostics of pathogens.

Results: For the first time, we demonstrate that a single Au-nanoprobe may provide for detection of two distinct targets (pathogens) allowing colorimetric multi-target detection. We demonstrate this concept by using one single gold-nanoprobe capable to detect members of the Mycobacterium tuberculosis complex and Plasmodium sp., the etiologic agents of tuberculosis and malaria, respectively. Following characterisation, the developed gold-nanoprobe allowed detection of either target in individual samples or in samples containing both DNA species with the same efficacy.

Conclusions: Using one single probe via the non-cross-linking colorimetric methodology it is possible to identify multiple targets in one sample in one reaction. This proof-of-concept approach may easily be integrated into sensing platforms allowing for fast and simple multiplexing of Au-nanoprobe based detection at point-of-need.

No MeSH data available.


Related in: MedlinePlus