Τίτλος:
A novel non-amplification assay for the detection of Leishmania spp. in clinical samples using gold nanoparticles
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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.
Συγγραφείς:
Andreadou, M.
Liandris, E.
Gazouli, M.
Taka, S.
Antoniou, M.
Theodoropoulos, G.
Tachtsidis, I.
Goutas, N.
Vlachodimitropoulos, D.
Kasampalidis, I.
Ikonomopoulos, J.
Περιοδικό:
Journal of Microbiological Methods
Λέξεις-κλειδιά:
complementary DNA; gold nanoparticle, animal experiment; article; blood sampling; controlled study; diagnostic test accuracy study; diagnostic value; DNA determination; Leishmania; leishmaniasis; limit of detection; nonhuman; oligonucleotide probe; parasite identification; particle size; polymerase chain reaction; priority journal; sensitivity and specificity, Diagnosis; DNA; Gold nanoparticles; Leishmania; Nanotechnology, Animals; Color; DNA, Kinetoplast; DNA, Protozoan; Dogs; Gold; Humans; Leishmania; Leishmaniasis; Molecular Diagnostic Techniques; Nanoparticles; Oligonucleotide Probes; Parasitology; Reproducibility of Results; Sensitivity and Specificity
DOI:
10.1016/j.mimet.2013.10.011
