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A novel non-amplification assay for the detection of Leishmania spp. in clinical samples using gold nanoparticles

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dc.contributor.author Andreadou, M en
dc.contributor.author Liandris, E en
dc.contributor.author Gazouli, M en
dc.contributor.author Taka, S en
dc.contributor.author Antoniou, M en
dc.contributor.author Theodoropoulos, G en
dc.contributor.author Tachtsidis, I en
dc.contributor.author Goutas, N en
dc.contributor.author Vlachodimitropoulos, D en
dc.contributor.author Kasampalidis, I en
dc.contributor.author Ikonomopoulos, J en
dc.date.accessioned 2014-06-06T06:52:56Z
dc.date.available 2014-06-06T06:52:56Z
dc.date.issued 2014 en
dc.identifier.issn 01677012 en
dc.identifier.uri http://dx.doi.org/10.1016/j.mimet.2013.10.011 en
dc.identifier.uri http://62.217.125.90/xmlui/handle/123456789/6264
dc.subject Diagnosis en
dc.subject DNA en
dc.subject Gold nanoparticles en
dc.subject Leishmania en
dc.subject Nanotechnology en
dc.subject.other complementary DNA en
dc.subject.other gold nanoparticle en
dc.subject.other animal experiment en
dc.subject.other article en
dc.subject.other blood sampling en
dc.subject.other controlled study en
dc.subject.other diagnostic test accuracy study en
dc.subject.other diagnostic value en
dc.subject.other DNA determination en
dc.subject.other Leishmania en
dc.subject.other leishmaniasis en
dc.subject.other limit of detection en
dc.subject.other nonhuman en
dc.subject.other oligonucleotide probe en
dc.subject.other parasite identification en
dc.subject.other particle size en
dc.subject.other polymerase chain reaction en
dc.subject.other priority journal en
dc.subject.other sensitivity and specificity en
dc.title A novel non-amplification assay for the detection of Leishmania spp. in clinical samples using gold nanoparticles en
heal.type journalArticle en
heal.identifier.primary 10.1016/j.mimet.2013.10.011 en
heal.publicationDate 2014 en
heal.abstract Leishmaniosis is a zoonose caused by protozoans of the genus Leishmania. The need for accurate diagnostic investigation of cases of leishmaniosis has rendered today the use of molecular biology techniques broadly applicable. However, the reliable application of these methods requires highly-specialised personnel, dedicated equipment and space. The aim of this study was the design and construction of functionalized gold nanoparticles (AuNPs) that would be incorporated into an easily applicable DNA detection methodology for the identification of Leishmania spp. in clinical samples.AuNPs 20. nm in diameter were conjugated with four oligonucleotide probes, targeting kinetoplastid minicircle DNA of Leishmania spp. In the absence of complimentary DNA, AuNPs-probes precipitate under acid environment causing a change of color from red to purple, which can be detected by visual observation. In the presence of target DNA the color of the solution remains red. The specific methodology was applied to positive and negative control samples and whole blood collected from dogs with suspected canine leishmaniosis. The method's minimum detection limit was defined to 11.5. ng of target DNA per μl of sample. Repeatability and reproducibility were 100%. Relative sensitivity and specificity referenced to PCR were calculated to 92% and 100% regarding collectively control and clinical samples.The proposed approach can be considered an appealing diagnostic solution especially for screening purposes in enzootic areas, where detection of very small amounts of the targeted analyte is not top priority. © 2013 Elsevier B.V. en
heal.journalName Journal of Microbiological Methods en
dc.identifier.issue 1 en
dc.identifier.volume 96 en
dc.identifier.doi 10.1016/j.mimet.2013.10.011 en
dc.identifier.spage 56 en
dc.identifier.epage 61 en


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