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Toldrà Filella, AnnaORCID iD iconorcid.org/0000-0001-6371-6055
Publications (10 of 10) Show all publications
Toldrà Filella, A., Chondrogiannis, G. & Hamedi, M. (2023). A 3D paper microfluidic device for enzyme-linked assays: Application to DNA analysis. Biotechnology Journal, 18(9), Article ID 2300143.
Open this publication in new window or tab >>A 3D paper microfluidic device for enzyme-linked assays: Application to DNA analysis
2023 (English)In: Biotechnology Journal, ISSN 1860-6768, E-ISSN 1860-7314, Vol. 18, no 9, article id 2300143Article in journal (Refereed) Published
Abstract [en]

A paper microfluidic device capable of conducting enzyme-linked assays is presented: a microfluidic enzyme-linked paper analytical device (μEL-PAD). The system exploits a wash-free sandwich coupling to form beads/analyte/enzyme complexes, which are subsequently added to the vertical flow device composed of wax-printed paper, waxed nitrocellulose membrane and absorbent/barrier layers. The nitrocellulose retains the bead complexes without disrupting the flow, enabling for an efficient washing step. The entrapped complexes then interact with the chromogenic substrate stored on the detection paper, generating a color change on it, quantified with an open-source smartphone software. This is a universal paper-based technology suitable for high-sensitivity quantification of many analytes, such as proteins or nucleic acids, with different enzyme-linked formats. Here, the potential of the μEL-PAD is demonstrated to detect DNA from Staphylococcus epidermidis. After generation of isothermally amplified genomic DNA from bacteria, Biotin/FITC-labeled products were analyzed with the μEL-PAD, exploiting streptavidin-coated beads and antiFITC-horseradish peroxidase. The μEL-PAD achieved a limit of detection (LOD) and quantification <10 genome copies/μL, these being at least 70- and 1000-fold lower, respectively, than a traditional lateral flow assay (LFA) exploiting immobilized streptavidin and antiFITC-gold nanoparticles. It is envisaged that the device will be a good option for low-cost, simple, quantitative, and sensitive paper-based point-of-care testing.

Place, publisher, year, edition, pages
Wiley, 2023
Keywords
high-sensitivity paper analytical device, lateral flow test, point-of-care diagnostics, quantitative multi-step assay, smartphone colorimetric readout, vertical flow μPAD
National Category
Polymer Chemistry
Identifiers
urn:nbn:se:kth:diva-338571 (URN)10.1002/biot.202300143 (DOI)000999468200001 ()37222181 (PubMedID)2-s2.0-85161407012 (Scopus ID)
Note

QC 20231107

Available from: 2023-11-07 Created: 2023-11-07 Last updated: 2023-11-07Bibliographically approved
Hanze, M., Khaliliazar, S., Reu, P., Toldrà Filella, A. & Hamedi, M. (2023). Toward Continuous Molecular Testing Using Gold-Coated Threads as Multi-Target Electrochemical Biosensors. Biosensors, 13(9), Article ID 844.
Open this publication in new window or tab >>Toward Continuous Molecular Testing Using Gold-Coated Threads as Multi-Target Electrochemical Biosensors
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2023 (English)In: Biosensors, ISSN 2079-6374, Vol. 13, no 9, article id 844Article in journal (Refereed) Published
Abstract [en]

Analytical systems based on isothermal nucleic acid amplification tests (NAATs) paired with electroanalytical detection enable cost-effective, sensitive, and specific digital pathogen detection for various in situ applications such as point-of-care medical diagnostics, food safety monitoring, and environmental surveillance. Self-assembled monolayers (SAMs) on gold surfaces are reliable platforms for electroanalytical DNA biosensors. However, the lack of automation and scalability often limits traditional chip-based systems. To address these challenges, we propose a continuous thread-based device that enables multiple electrochemical readings on a functionalized working electrode Au thread with a single connection point. We demonstrate the possibility of rolling the thread on a spool, which enables easy manipulation in a roll-to-roll architecture for high-throughput applications. As a proof of concept, we have demonstrated the detection of recombinase polymerase amplification (RPA) isothermally amplified DNA from the two toxic microalgae species Ostreopsis cf. ovata and Ostreopsis cf. siamensis by performing a sandwich hybridization assay (SHA) with electrochemical readout.

Place, publisher, year, edition, pages
MDPI AG, 2023
Keywords
chronoamperometry, isothermal DNA amplification, metal-coated threads, roll-to-roll, sandwich hybridization assay, self-assembled monolayers
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:kth:diva-338402 (URN)10.3390/bios13090844 (DOI)001074469800001 ()37754078 (PubMedID)2-s2.0-85172180625 (Scopus ID)
Note

QC 20231023

Available from: 2023-10-23 Created: 2023-10-23 Last updated: 2023-10-24Bibliographically approved
Toldrà Filella, A., Ainla, A., Khaliliazar, S., Landin, R., Chondrogiannis, G., Hanze, M., . . . Hamedi, M. (2022). Portable electroanalytical nucleic acid amplification tests using printed circuit boards and open-source electronics. The Analyst, 147(19), 4249-4256
Open this publication in new window or tab >>Portable electroanalytical nucleic acid amplification tests using printed circuit boards and open-source electronics
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2022 (English)In: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 147, no 19, p. 4249-4256Article in journal (Refereed) Published
Abstract [en]

The realization of electrochemical nucleic acid amplification tests (NAATs) at the point of care (POC) is highly desirable, but it remains a challenge given their high cost and lack of true portability/miniaturization. Here we show that mass-produced, industrial standardized, printed circuit boards (PCBs) can be repurposed to act as near-zero cost electrodes for self-assembled monolayer-based DNA biosensing, and further integration with a custom-designed and low-cost portable potentiostat. To show the analytical capability of this system, we developed a NAAT using isothermal recombinase polymerase amplification, bypassing the need of thermal cyclers, followed by an electrochemical readout relying on a sandwich hybridization assay. We used our sensor and device for analytical detection of the toxic microalgae Ostreopsis cf. ovata as a proof of concept. This work shows the potential of PCBs and open-source electronics to be used as powerful POC DNA biosensors at a low-cost. 

Place, publisher, year, edition, pages
Royal Society of Chemistry (RSC), 2022
Keywords
Biosensing Techniques, DNA, Electronics, Nucleic Acid Amplification Techniques, Recombinases, Costs, Nucleic acids, Self assembled monolayers, Timing circuits, Voltage regulators, recombinase, DNA biosensing, Electrochemicals, High costs, Low-costs, Miniaturisation, Nucleic acids amplification, Open-source, Point of care, Potentiostats, genetic procedures, genetics, Printed circuit boards
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:kth:diva-327273 (URN)10.1039/d2an00923d (DOI)000842617500001 ()35993403 (PubMedID)2-s2.0-85138024123 (Scopus ID)
Note

QC 20230523

Available from: 2023-05-23 Created: 2023-05-23 Last updated: 2023-05-23Bibliographically approved
Leonardo, S., Toldrà Filella, A. & Campas, M. (2021). Biosensors Based on Isothermal DNA Amplification for Bacterial Detection in Food Safety and Environmental Monitoring. Sensors, 21(2), Article ID 602.
Open this publication in new window or tab >>Biosensors Based on Isothermal DNA Amplification for Bacterial Detection in Food Safety and Environmental Monitoring
2021 (English)In: Sensors, E-ISSN 1424-8220, Vol. 21, no 2, article id 602Article, review/survey (Refereed) Published
Abstract [en]

The easy and rapid spread of bacterial contamination and the risk it poses to human health makes evident the need for analytical methods alternative to conventional time-consuming laboratory-based techniques for bacterial detection. To tackle this demand, biosensors based on isothermal DNA amplification methods have emerged, which avoid the need for thermal cycling, thus facilitating their integration into small and low-cost devices for in situ monitoring. This review focuses on the breakthroughs made on biosensors based on isothermal amplification methods for the detection of bacteria in the field of food safety and environmental monitoring. Optical and electrochemical biosensors based on loop mediated isothermal amplification (LAMP), rolling circle amplification (RCA), recombinase polymerase amplification (RPA), helicase dependent amplification (HDA), strand displacement amplification (SDA), and isothermal strand displacement polymerisation (ISDPR) are described, and an overview of their current advantages and limitations is provided. Although further efforts are required to harness the potential of these emerging analytical techniques, the coalescence of the different isothermal amplification techniques with the wide variety of biosensing detection strategies provides multiple possibilities for the efficient detection of bacteria far beyond the laboratory bench.

Place, publisher, year, edition, pages
MDPI AG, 2021
Keywords
bacteria, biosensor, isothermal DNA amplification, food safety, environmental monitoring, loop mediated isothermal amplification (LAMP), rolling circle amplification (RCA), recombinase polymerase amplification (RPA), helicase dependent amplification (HDA), strand displacement amplification (SDA), isothermal strand displacement polymerisation (ISDPR)
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-290495 (URN)10.3390/s21020602 (DOI)000611700300001 ()33467078 (PubMedID)2-s2.0-85099515850 (Scopus ID)
Note

QC 20210304.

Available from: 2021-03-04 Created: 2021-03-04 Last updated: 2022-06-25Bibliographically approved
Khaliliazar, S., Toldrà, A., Chondrogiannis, G. & Hamedi, M. (2021). Electroanalytical Paper-Based Nucleic Acid Amplification Biosensors with Integrated Thread Electrodes. Analytical Chemistry, 93(42), 14187-14195
Open this publication in new window or tab >>Electroanalytical Paper-Based Nucleic Acid Amplification Biosensors with Integrated Thread Electrodes
2021 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 93, no 42, p. 14187-14195Article in journal (Refereed) Published
Abstract [en]

Nucleic acid amplification tests (NAATs) are very sensitive and specific methods, but they mainly rely on centralized laboratories and therefore are not suitable for point-of-care testing. Here, we present a 3D microfluidic paper-based electrochemical NAAT. These devices use off-the-shelf gold plasma-coated threads to integrate electroanalytical readouts using ex situ self-assembled monolayer formation on the threads prior to assembling into the paper device. They further include a sandwich hybridization assay with sample incubation, rinsing, and detection steps all integrated using movable stacks of filter papers to allow time-sequenced reactions. The devices use glass fiber substrates for storing recombinase polymerase amplification reagents and conducting the isothermal amplification. We used the paper-based device for the detection of the toxic microalgae Ostreopsis cf. ovata. The NAAT, completed in 95 min, attained a limit of detection of 0.06 pM target synthetic DNA and was able to detect 1 ng/mu L O. cf. ovata genomic DNA with negligible cross-reactivity from a closely related microalgae species. We think that the integration of thread electrodes within paper-based devices paves the way for digital one-time use NAATs and numerous other advanced electroanalytical paper- or textile-based devices.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2021
National Category
Infectious Medicine
Identifiers
urn:nbn:se:kth:diva-304780 (URN)10.1021/acs.analchem.1c02900 (DOI)000711718700022 ()34648274 (PubMedID)2-s2.0-85118249212 (Scopus ID)
Note

QC 20211118

Available from: 2021-11-18 Created: 2021-11-18 Last updated: 2024-03-15Bibliographically approved
Chondrogiannis, G., Khaliliazar, S., Toldrà Filella, A., Reu, P. & Hamedi, M. (2021). Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests. Scientific Reports, 11(1), Article ID 6140.
Open this publication in new window or tab >>Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
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2021 (English)In: Scientific Reports, E-ISSN 2045-2322, Vol. 11, no 1, article id 6140Article in journal (Refereed) Published
Abstract [en]

Enzymes are the cornerstone of modern biotechnology. Achromopeptidase (ACP) is a well-known enzyme that hydrolyzes a number of proteins, notably proteins on the surface of Gram-positive bacteria. It is therefore used for sample preparation in nucleic acid tests. However, ACP inhibits DNA amplification which makes its integration difficult. Heat is commonly used to inactivate ACP, but it can be challenging to integrate heating into point-of-care devices. Here, we use recombinase polymerase amplification (RPA) together with ACP, and show that when ACP is immobilized on nitrocellulose paper, it retains its enzymatic function and can easily and rapidly be activated using agitation. The nitrocellulose-bound ACP does, however, not leak into the solution, preventing the need for deactivation through heat or by other means. Nitrocellulose-bound ACP thus opens new possibilities for paper-based Point-of-Care (POC) devices.

Place, publisher, year, edition, pages
Springer Nature, 2021
National Category
Biological Sciences
Identifiers
urn:nbn:se:kth:diva-292300 (URN)10.1038/s41598-021-85481-2 (DOI)000630510600002 ()33731748 (PubMedID)2-s2.0-85102733962 (Scopus ID)
Note

QC 20210406

Available from: 2021-04-06 Created: 2021-04-06 Last updated: 2022-11-03Bibliographically approved
Toldrà Filella, A., O'Sullivan, C. K., Diogene, J. & Campas, M. (2020). Detecting harmful algal blooms with nucleic acid amplification-based biotechnological tools. Science of the Total Environment, 749, Article ID 141605.
Open this publication in new window or tab >>Detecting harmful algal blooms with nucleic acid amplification-based biotechnological tools
2020 (English)In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 749, article id 141605Article, review/survey (Refereed) Published
Abstract [en]

Harmful algal blooms (HABs) represent a growing threat to aquatic ecosystems and humans. Effective HAB management and mitigation efforts strongly rely on the availability of timely and in-situ tools for the detection of microalgae. In this sense, nudeic acid-based (molecular) methods are being considered for the unequivocal identification of miaoalgae as an attractive alternative to the currently used time-consuming and laboratory-based light microscopy techniques. This review provides an overview of the progress made on new molecular biotechnological tools for microalgal detection, particularly focusing on those that combine a nudeic acid (DNA or RNA) amplification step with detection. Different types of amplification processes (thermal and isothermal) and detection formats (e.g. microarrays, biosensors, lateral flows) are presented, and a comprehensive overview of their advantages and limitations is provided Although isothermal techniques are an attractive alternative to thermal amplification to reach in-situ analysis, further development is still required. Finally, current challenges, critical steps and future directions of the whole analysis process ( from sample procurement to in-situ implementation) are described.

Place, publisher, year, edition, pages
Elsevier BV, 2020
Keywords
Harmful algal blooms (HABs), Microalgae, Molecular method, Isothermal DNA amplification, In-situ HAB monitoring
National Category
Ecology
Identifiers
urn:nbn:se:kth:diva-287520 (URN)10.1016/j.scitotenv.2020.141605 (DOI)000581793800091 ()32827817 (PubMedID)2-s2.0-85089472355 (Scopus ID)
Note

QC 20201218

Available from: 2020-12-18 Created: 2020-12-18 Last updated: 2022-06-25Bibliographically approved
Toldrà Filella, A., Chondrogiannis, G. & Hamedi, M.3D paper microfluidic devices for enzyme‐linked assays, and itsapplication to DNA analysis.
Open this publication in new window or tab >>3D paper microfluidic devices for enzyme‐linked assays, and itsapplication to DNA analysis
(English)Manuscript (preprint) (Other academic)
National Category
Paper, Pulp and Fiber Technology Biomaterials Science Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-320959 (URN)
Note

QC 20221109

Available from: 2022-11-03 Created: 2022-11-03 Last updated: 2022-11-09Bibliographically approved
Khaliliazar, S. Electroanalytical Paper based Nucleic Acid Amplification Tests with Integrated Thread Electrodes.
Open this publication in new window or tab >>Electroanalytical Paper based Nucleic Acid Amplification Tests with Integrated Thread Electrodes
(English)Article in journal (Refereed) [Artistic work] Submitted
National Category
Natural Sciences
Identifiers
urn:nbn:se:kth:diva-302112 (URN)
Funder
EU, European Research Council
Note

QC 20210923

Available from: 2021-09-16 Created: 2021-09-16 Last updated: 2022-11-03Bibliographically approved
Chondrogiannis, G., Toldrà Filella, A. & Hanze, M.Paper‐based RNase digestion towards viral nucleic acid self‐tests.
Open this publication in new window or tab >>Paper‐based RNase digestion towards viral nucleic acid self‐tests
(English)Manuscript (preprint) (Other academic)
National Category
Paper, Pulp and Fiber Technology Medical Materials Medical Biotechnology
Identifiers
urn:nbn:se:kth:diva-320957 (URN)
Note

QC 20221107

Available from: 2022-11-03 Created: 2022-11-03 Last updated: 2022-11-07Bibliographically approved
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Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0001-6371-6055

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