title: Gold nanoparticle-based lateral flow biosensor for rapid visual detection of Leishmania-specific DNA amplification products.
abstract: Leishmaniasis is a disease, caused by Leishmania parasites, which infect humans and animals, posing a major social and economic burden worldwide. The need for accurate and sensitive disease diagnosis led to the widespread adoption of PCR amplification. Detection of the amplification products (i.e. gel electrophoresis) require time-consuming protocols performed by trained personnel, with high cost. Aim of the present study was the simplification of PCR product detection, using a nucleic acid lateral flow, combined with functionalized gold nanoparticles. Amplification reactions targeting kinetoplastid DNA of Leishmania spp were performed on canine blood samples and a positive signal was formed as a red test zone. The visual detection was completed in 20min. Extensive optimization enabled the detection of 100fmol of target DNA. Clinical samples of infected dog blood were analyzed with high specificity. Overall, the proposed lateral flow biosensor can be considered an appealing alternative platform for Leishmania-specific amplification products detection with low cost and attractive simplicity.
authors.affiliation: Laboratory of Cellular Immunology, Department of Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias Ave., 11521 Athens, Greece. Electronic address: dtouban@pasteur.gr.
authors.forename: Dimitra K
authors.initials: DK
authors.lastname: Toubanaki
authors.affiliation: Laboratory of Cellular Immunology, Department of Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias Ave., 11521 Athens, Greece. Electronic address: eathanasiou@pasteur.gr.
authors.forename: Evita
authors.initials: E
authors.lastname: Athanasiou
authors.affiliation: Laboratory of Cellular Immunology, Department of Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias Ave., 11521 Athens, Greece. Electronic address: ekaragouni@pasteur.gr.
authors.forename: Evdokia
authors.initials: E
authors.lastname: Karagouni
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D018105:DNA, Kinetoplast / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification*; D004285:Dogs; D006046:Gold; D007891:Leishmania / Q000235:genetics*; D007896:Leishmaniasis / Q000175:diagnosis / Q000469:parasitology; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D006046:Gold
keywords: Gold nanoparticles; Kinetoplast gene; Lateral flow strip biosensor; Leishmania