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Evaluation of the performance of selected in-house and commercially available PCR and real-time PCR assays for the detection of Leishmania DNA in canine clinical samples

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dc.contributor.author Andreadou, M en
dc.contributor.author Liandris, E en
dc.contributor.author Kasampalidis, IN en
dc.contributor.author Taka, S en
dc.contributor.author Antoniou, M en
dc.contributor.author Ntais, P en
dc.contributor.author Vaiopoulou, A en
dc.contributor.author Theodoropoulos, G en
dc.contributor.author Gazouli, M en
dc.contributor.author Ikonomopoulos, J en
dc.date.accessioned 2014-06-06T06:51:48Z
dc.date.available 2014-06-06T06:51:48Z
dc.date.issued 2012 en
dc.identifier.issn 00144894 en
dc.identifier.uri http://dx.doi.org/10.1016/j.exppara.2012.05.012 en
dc.identifier.uri http://62.217.125.90/xmlui/handle/123456789/5701
dc.subject Leishmania en
dc.subject Molecular detection en
dc.subject PCR en
dc.subject.other protozoal DNA en
dc.subject.other article en
dc.subject.other comparative study en
dc.subject.other controlled study en
dc.subject.other DNA sequence en
dc.subject.other dog en
dc.subject.other human en
dc.subject.other Leishmania en
dc.subject.other leishmaniasis en
dc.subject.other limit of detection en
dc.subject.other microorganism detection en
dc.subject.other nonhuman en
dc.subject.other nucleotide sequence en
dc.subject.other parasite isolation en
dc.subject.other polymerase chain reaction en
dc.subject.other priority journal en
dc.subject.other promastigote en
dc.subject.other real time polymerase chain reaction en
dc.subject.other reproducibility en
dc.subject.other sensitivity and specificity en
dc.subject.other statistical analysis en
dc.subject.other Animals en
dc.subject.other DNA Primers en
dc.subject.other DNA, Protozoan en
dc.subject.other Dog Diseases en
dc.subject.other Dogs en
dc.subject.other Fluorescent Antibody Technique, Indirect en
dc.subject.other Humans en
dc.subject.other Leishmania en
dc.subject.other Leishmaniasis en
dc.subject.other Limit of Detection en
dc.subject.other Polymerase Chain Reaction en
dc.subject.other Real-Time Polymerase Chain Reaction en
dc.subject.other Reproducibility of Results en
dc.subject.other Sensitivity and Specificity en
dc.subject.other Protozoa en
dc.title Evaluation of the performance of selected in-house and commercially available PCR and real-time PCR assays for the detection of Leishmania DNA in canine clinical samples en
heal.type journalArticle en
heal.identifier.primary 10.1016/j.exppara.2012.05.012 en
heal.publicationDate 2012 en
heal.abstract Protozoa of the genus Leishmania are the causative agents of leishmaniosis. Although the polymerase chain reaction (PCR) has proved very effective in the detection of Leishmania DNA, a standardized method does not exist. In this study we attempt a comparative evaluation between one real time PCR (Method D), two in-house (Methods A and C), and a commercially available PCR assay (Method B) for the detection of Leishmania DNA, in order to support reliable diagnostic investigation of leishmaniosis. This evaluation was performed in regard to relative specificity and sensitivity, minimum detection limit (MDL), repeatability and reproducibility using cultured isolates and clinical samples. All the methods under study produced the expected result with the positive and negative controls. However with regard to clinical samples, Method C showed a statistically significant higher level of positivity. Relative sensitivity and specificity of Methods A, B and D in comparison to C was calculated respectively at 50.7%, 43%, 40%, and 90.8%, 93.4% and 89.5%. The MDL for Methods A-D was defined respectively at 30.7, 5, 3.7, and 5 promastigotes/ml. Repeatability and reproducibility were excellent in all cases with only the exception of Method A regarding reproducibility with a different brand of PCR reagents. The results that were recorded indicate that evaluation of PCR assays before their application for research and clinical diagnosis can provide useful evidence for their reliable application. Within this context the use of internal amplification controls and the confirmation of the specificity of the amplification product is recommended. © 2012 Elsevier Inc. en
heal.journalName Experimental Parasitology en
dc.identifier.issue 4 en
dc.identifier.volume 131 en
dc.identifier.doi 10.1016/j.exppara.2012.05.012 en
dc.identifier.spage 419 en
dc.identifier.epage 424 en


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