The Experts below are selected from a list of 252 Experts worldwide ranked by ideXlab platform
Panagiotis Karanis - One of the best experts on this subject based on the ideXlab platform.
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evaluation of loop mediated isothermal amplification for detection of toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and Immunofluorescence Test ift
Diagnostic Microbiology and Infectious Disease, 2008Co-Authors: Isaia Sotiriadou, Panagiotis KaranisAbstract:The development and evaluation of a 1-step single-tube accelerated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Toxoplasma in water samples is described. The method has been evaluated based on the amplification of B1 and TgOWP Toxoplasma genes, and it demonstrated a sensitivity detection limit of 0.1 tachyzoites' DNA for both genes. LAMP detection was evaluated and compared with nested polymerase chain reaction (PCR) in 26 water sample pellets spiked with known numbers of Toxoplasma oocysts. After DNA extraction, the detection sensitivity in spiked pellets was 100% by LAMP and 53.8% by PCR. Subsequently, 52 natural water samples of different origin were directly investigated by 3 assays: LAMP, PCR, and Immunofluorescence Test (IFT). Twenty-five (48%) of 52 have been found positive for Toxoplasma DNA by LAMP, whereas nested PCR products were generated in 7 of 52 (13.5%) water samples. All 52 water samples were negative for Toxoplasma by IFT. These data clearly indicate LAMP as a rapid, specific, and sensitive tool for the detection of Toxoplasma contamination in water samples.
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a comparison of phase contrast microscopy and an Immunofluorescence Test for the detection of giardia spp in faecal specimens from cattle and wild rodents
Transactions of The Royal Society of Tropical Medicine and Hygiene, 1996Co-Authors: Panagiotis Karanis, K Opiela, M Alarousi, Hanns M SeitzAbstract:As part of a study on the presence of Giardia cysts in water, a commercial direct Immunofluorescence (DIF) Test for the cysts was evaluated and compared to phase contrast microscopy. Forty faecal samples were collected from cattle; 31 contained Giardia. All 31 samples were positive by DIF whereas only 17 were detected by microscopy. The detected Giardia cysts were identified as belonging to the G. duodenalis group. 216 faecal samples were collected from wild rodents (Apodemus flavicollis, A. sylvaticus and Clethrionomys glareolus); 103 contained Giardia. Cysts were detected in 97 samples by DIF and in 57 by microscopy; 51 samples were positive by both methods. Nineteen rodents harboured cysts which morphologically resembled G. duodenalis. Specific identification was possible only by phase contrast microscopy, but the results suggested that DIF was superior for the detection of Giardia cysts in faeces of the animals Tested.
Paolo Soda - One of the best experts on this subject based on the ideXlab platform.
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the inter observer reading variability in anti nuclear antibodies indirect ana Immunofluorescence Test a multicenter evaluation and a review of the literature
Autoimmunity Reviews, 2017Co-Authors: Amelia Rigon, Maria Infantino, Giulio Iannello, Mariangela Manfredi, Mario Merone, Angela Tincani, Ilaria Cavazzana, N Carabellese, Antonella Radice, Paolo SodaAbstract:Recently there has been an increase demand for Computer-Aided Diagnosis (CAD) tools to support clinicians in the field of Indirect Immunofluorescence (IIF), as the novel digital imaging reading approach can help to overcome the reader subjectivity. Nevertheless, a large multicenter evaluation of the inter-observer reading variability in this field is still missing. This work fills this gap as we evaluated 556 consecutive samples, for a total of 1679 images, collected in three laboratories with IIF expertise using HEp-2 cell substrate (MBL) at 1:80 screening dilution according to conventional procedures. In each laboratory, the images were blindly classified by two experts into three intensity classes: positive, negative, and weak positive. Positive and weak positive ANA-IIF results were categorized by the predominant fluorescence pattern among six main classes. Data were pairwise analyzed and the inter-observer reading variability was measured by Cohen's kappa Test, revealing a pairwise agreement little further away than substantial both for fluorescence intensity and for staining pattern recognition (k=0.602 and k=0.627, respectively). We also noticed that the inter-observer reading variability decreases when it is measured with respect to a gold standard classification computed on the basis of labels assigned by the three laboratories. These data show that laboratory agreement improves using digital images and comparing each single human evaluation to potential reference data, suggesting that a solid gold standard is essential to properly make use of CAD systems in routine work lab.
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the burden of the variability introduced by the hep 2 assay kit and the cad system in ana indirect Immunofluorescence Test
Immunologic Research, 2017Co-Authors: Maria Infantino, Francesca Meacci, Valentina Grossi, Mariangela Manfredi, Maurizio Benucci, Mario Merone, Paolo SodaAbstract:According to the recent recommendations of the American College of Rheumatology, ANA Task Force, IIF technique should be considered the gold standard in antinuclear antibodies (ANAs) Testing. To overcome the lack of standardization, biomedical industries have developed several computer-aided diagnosis (CAD) systems. Two hundred and sixty-one consecutive samples with suspected autoimmune diseases were Tested for ANA by means of IIF on routinely HEp-2 assay kit (Euroimmun AG). Assignment of result was made if consensus for positive/negative was reached by at least 2 out of 3 expert physicians. ANA-IIF was also carried out using 3 CAD systems: Zenit G-Sight (n = 84), Helios (n = 85) and NOVA View (n = 92); human evaluation was repeated on the same substrate of each CAD system (Immco, Aesku and Inova HEp-2 cells, respectively). To anonymize the results, we randomly named these three systems as A, B and C. We ran a statistical analysis computing several measures of agreement between the ratings, and we also improved the evaluation by using the Wilcoxon’s Test for nonparametric data. Agreement between the human readings on routinely HEp-2 assay kit and human readings on CAD HEp-2 assay was substantial for A (k = 0.82) and B (k = 0.72), and almost perfect for C (k = 0.89). Such readings were statistically different only in case A. Comparing experts’ readings with the readings of CAD systems, when the samples were prepared using CAD HEp-2 assay kits, we found almost perfect agreement for B and C (k = 0.86; k = 0.82) and substantial agreement for A (k = 0.73). Again, human and CAD readings were statistically different only in A. When we compared the readings of medical experts on routinely HEp-2 assay kit with the output of the CAD systems that worked using their own slides, we found substantial agreement for all the systems (A: k = 0.62; B: k = 0.65; C: k = 0.71). Such readings were not statistically different. The change of the assay kit and/or the introduction of a CAD system affect the laboratory reporting, with an evident impact on the autoimmune laboratory workflow. The CAD systems may represent one of the most important novel elements of harmonization in the autoimmunity field, reducing intra- and inter-laboratory variability in a new vision of the diagnostic autoimmune platform.
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the classification of crithidia luciliae Immunofluorescence Test clift using a novel automated system
Arthritis Research & Therapy, 2014Co-Authors: F Buzzulini, Amelia Rigon, Paolo Soda, Leonardo Onofri, Maria Infantino, L Arcarese, Giulio Iannello, Antonella AfeltraAbstract:Introduction In recent years, there has been an increased demand for computer-aided diagnosis (CAD) tools to support clinicians in the field of indirect Immunofluorescence. To this aim, academic and industrial research is focusing on detecting antinuclear, anti-neutrophil, and anti-double-stranded (anti-dsDNA) antibodies. Within this framework, we present a CAD system for automatic analysis of dsDNA antibody images using a multi-step classification approach. The final classification of a well is based on the classification of all its images, and each image is classified on the basis of the labeling of its cells.
Isaia Sotiriadou - One of the best experts on this subject based on the ideXlab platform.
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evaluation of loop mediated isothermal amplification for detection of toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and Immunofluorescence Test ift
Diagnostic Microbiology and Infectious Disease, 2008Co-Authors: Isaia Sotiriadou, Panagiotis KaranisAbstract:The development and evaluation of a 1-step single-tube accelerated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Toxoplasma in water samples is described. The method has been evaluated based on the amplification of B1 and TgOWP Toxoplasma genes, and it demonstrated a sensitivity detection limit of 0.1 tachyzoites' DNA for both genes. LAMP detection was evaluated and compared with nested polymerase chain reaction (PCR) in 26 water sample pellets spiked with known numbers of Toxoplasma oocysts. After DNA extraction, the detection sensitivity in spiked pellets was 100% by LAMP and 53.8% by PCR. Subsequently, 52 natural water samples of different origin were directly investigated by 3 assays: LAMP, PCR, and Immunofluorescence Test (IFT). Twenty-five (48%) of 52 have been found positive for Toxoplasma DNA by LAMP, whereas nested PCR products were generated in 7 of 52 (13.5%) water samples. All 52 water samples were negative for Toxoplasma by IFT. These data clearly indicate LAMP as a rapid, specific, and sensitive tool for the detection of Toxoplasma contamination in water samples.
Danilo Villalta - One of the best experts on this subject based on the ideXlab platform.
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Diagnosing systemic lupus erythematosus: New-generation immunoassays for measurement of anti-dsDNA antibodies are an effective alternative to the Farr technique and the Crithidia luciliae Immunofluorescence Test
Lupus, 2010Co-Authors: Andrés Antico, S. Platzgummer, Danila Bassetti, Renato Tozzoli, Nicola Bizzaro, Danilo VillaltaAbstract:The aim of this study was to evaluate the diagnostic performance of four new enzyme immunoassays (EIAs) for anti-double-stranded-DNA (anti-dsDNA) antibodies, in comparison with the Farr assay and the Crithidia luciliae Immunofluorescence Test (CLIFT). To this purpose, sera from four patient groups were collected: 52 sera from patients with systemic lupus erythematosus (SLE); 28 from patients with other connective tissue diseases (CTD); 36 from patients with hepatitis C virus (HCV) infection; and 24 from those with acute viral infection. All sera were Tested for anti-dsDNA antibodies by four EIA methods using a different antigenic DNA source [synthetic oligonucleotide (Method A), circular plasmid (Method B), recombinant (Method C), and purified extracted (Method D)], and by CLIFT and Farr assays. The diagnostic sensitivity of the assays was as follows: 84.6% (Method A), 73% (B), 82.7% (C), 84.6% (D), 55.8% (CLIFT), and 78.8% (Farr). Specificity was 82.9% (A), 97.7% (B), 96.5% (C), 94.3% (D), 96.5% (CLIFT), and 90.9% (Farr). From these data, we can conclude that the new-generation EIA methods evaluated in this study have higher sensitivity than the CLIFT and Farr assays and, with the exception of Method A, have specificity similar to the CLIFT and slightly higher than the Farr assay. These findings suggest that EIA Tests may replace CLIFT as a screening Test and the Farr assay as a specific Test, for anti-dsDNA antibody detection.
Maria Infantino - One of the best experts on this subject based on the ideXlab platform.
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the inter observer reading variability in anti nuclear antibodies indirect ana Immunofluorescence Test a multicenter evaluation and a review of the literature
Autoimmunity Reviews, 2017Co-Authors: Amelia Rigon, Maria Infantino, Giulio Iannello, Mariangela Manfredi, Mario Merone, Angela Tincani, Ilaria Cavazzana, N Carabellese, Antonella Radice, Paolo SodaAbstract:Recently there has been an increase demand for Computer-Aided Diagnosis (CAD) tools to support clinicians in the field of Indirect Immunofluorescence (IIF), as the novel digital imaging reading approach can help to overcome the reader subjectivity. Nevertheless, a large multicenter evaluation of the inter-observer reading variability in this field is still missing. This work fills this gap as we evaluated 556 consecutive samples, for a total of 1679 images, collected in three laboratories with IIF expertise using HEp-2 cell substrate (MBL) at 1:80 screening dilution according to conventional procedures. In each laboratory, the images were blindly classified by two experts into three intensity classes: positive, negative, and weak positive. Positive and weak positive ANA-IIF results were categorized by the predominant fluorescence pattern among six main classes. Data were pairwise analyzed and the inter-observer reading variability was measured by Cohen's kappa Test, revealing a pairwise agreement little further away than substantial both for fluorescence intensity and for staining pattern recognition (k=0.602 and k=0.627, respectively). We also noticed that the inter-observer reading variability decreases when it is measured with respect to a gold standard classification computed on the basis of labels assigned by the three laboratories. These data show that laboratory agreement improves using digital images and comparing each single human evaluation to potential reference data, suggesting that a solid gold standard is essential to properly make use of CAD systems in routine work lab.
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the burden of the variability introduced by the hep 2 assay kit and the cad system in ana indirect Immunofluorescence Test
Immunologic Research, 2017Co-Authors: Maria Infantino, Francesca Meacci, Valentina Grossi, Mariangela Manfredi, Maurizio Benucci, Mario Merone, Paolo SodaAbstract:According to the recent recommendations of the American College of Rheumatology, ANA Task Force, IIF technique should be considered the gold standard in antinuclear antibodies (ANAs) Testing. To overcome the lack of standardization, biomedical industries have developed several computer-aided diagnosis (CAD) systems. Two hundred and sixty-one consecutive samples with suspected autoimmune diseases were Tested for ANA by means of IIF on routinely HEp-2 assay kit (Euroimmun AG). Assignment of result was made if consensus for positive/negative was reached by at least 2 out of 3 expert physicians. ANA-IIF was also carried out using 3 CAD systems: Zenit G-Sight (n = 84), Helios (n = 85) and NOVA View (n = 92); human evaluation was repeated on the same substrate of each CAD system (Immco, Aesku and Inova HEp-2 cells, respectively). To anonymize the results, we randomly named these three systems as A, B and C. We ran a statistical analysis computing several measures of agreement between the ratings, and we also improved the evaluation by using the Wilcoxon’s Test for nonparametric data. Agreement between the human readings on routinely HEp-2 assay kit and human readings on CAD HEp-2 assay was substantial for A (k = 0.82) and B (k = 0.72), and almost perfect for C (k = 0.89). Such readings were statistically different only in case A. Comparing experts’ readings with the readings of CAD systems, when the samples were prepared using CAD HEp-2 assay kits, we found almost perfect agreement for B and C (k = 0.86; k = 0.82) and substantial agreement for A (k = 0.73). Again, human and CAD readings were statistically different only in A. When we compared the readings of medical experts on routinely HEp-2 assay kit with the output of the CAD systems that worked using their own slides, we found substantial agreement for all the systems (A: k = 0.62; B: k = 0.65; C: k = 0.71). Such readings were not statistically different. The change of the assay kit and/or the introduction of a CAD system affect the laboratory reporting, with an evident impact on the autoimmune laboratory workflow. The CAD systems may represent one of the most important novel elements of harmonization in the autoimmunity field, reducing intra- and inter-laboratory variability in a new vision of the diagnostic autoimmune platform.
-
the classification of crithidia luciliae Immunofluorescence Test clift using a novel automated system
Arthritis Research & Therapy, 2014Co-Authors: F Buzzulini, Amelia Rigon, Paolo Soda, Leonardo Onofri, Maria Infantino, L Arcarese, Giulio Iannello, Antonella AfeltraAbstract:Introduction In recent years, there has been an increased demand for computer-aided diagnosis (CAD) tools to support clinicians in the field of indirect Immunofluorescence. To this aim, academic and industrial research is focusing on detecting antinuclear, anti-neutrophil, and anti-double-stranded (anti-dsDNA) antibodies. Within this framework, we present a CAD system for automatic analysis of dsDNA antibody images using a multi-step classification approach. The final classification of a well is based on the classification of all its images, and each image is classified on the basis of the labeling of its cells.
