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001-es BibID:	BIBFORM072747
Első szerző:	Morton, C. Oliver
Cím:	Determining the analytical specificity of PCR-based assays for the diagnosis of IA : what is Aspergillus? / C. Oliver Morton, P. Lewis White, Rosemary A. Barnes, Lena Klingspor, Manuel Cuenca-Estrella, Katrien Lagrou, Stéphane Bretagne, Willem Melchers, Carlo Mengoli, Angela M. Caliendo, Massimo Cogliati, Yvette Debets-Ossenkopp, Rebecca Gorton, Ferry Hagen, Catriona Halliday, Petr Hamal, Kathleen Harvey-Wood, Katia Jaton, Gemma Johnson, Sarah Kidd, Martina Lengerova, Cornelia Lass-Florl, Chris Linton, Laurence Millon, C. Orla Morrissey, Melinda Paholcsek, Alida Fe Talento, Markus Ruhnke, Birgit Willinger, J. Peter Donnelly, Juergen Loeffler, EAPCRI
Dátum:	2017
ISSN:	1369-3786
Megjegyzések:	A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Aspergillus PCR analytical specificity cross reactivity detection range
Megjelenés:	Medical Mycology 55 : 4 (2017), p. 402-413. -
További szerzők:	White, P. Lewis Barnes, Rosemary A. Klingspor, Lena Cuenca-Estrella, Manuel Lagrou, Katrien Bretagne, Stéphane Melchers, Willem Mengoli, Carlo Caliendo, Angela M. Cogliati, Massimo Debets-Ossenkopp, Yvette Gorton, Rebecca Hagen, Ferry Halliday, Catriona Hamal, Petr Harvey-Wood, Kathleen Jaton, Katia Johnson, Gemma Kidd, Sarah Lengerova, Martina Lass-Florl, Cornelia Linton, Chris Millon, Laurence Morrissey, C. Orla Paholcsek Melinda (1984-) (molekuláris biológus, genetikus) Talento, Alida Fe Ruhnke, Markus Willinger, Birgit Donnelly, J. Peter Loeffler, Juergen EAPCRI
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001-es BibID:	BIBFORM053143
Első szerző:	Springer, Jan
Cím:	Multicenter comparison of serum and whole-blood specimens for detection of Aspergillus DNA in high-risk hematological patients / Jan Springer, C. O. Morton, Michael Perry, Werner J. Heinz, Melinda Paholcsek, Mona Alzheimer, T. R. Rogers, Rosemary A. Barnes, Hermann Einsele, Juergen Loeffler, P. Lewis White
Dátum:	2013
ISSN:	0095-1137
Megjegyzések:	Samples from patients at high risk for invasive aspergillosis (IA) were prospectively collected and analyzed for the presence of molecular markers of fungal infection. Serum specimens were screened for galactomannan and Aspergillus DNA, and whole-blood specimens were screened only for Aspergillus DNA. Fungal infections were categorized according to the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group, National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria. Forty-seven cases (proven and probable IA) and 31 controls (no evidence of IA) were selected retrospectively for this case-control study, comprising 803 samples, in order to determine the performance of whole-blood PCR, serum PCR, and serum galactomannan testing. Although no single assay was able to detect every case of IA, a combination of different assays provided the best performance. There was no significant difference between the use of whole-blood and serum specimens for PCR-based diagnosis of IA, but there was a trend for whole blood to be more sensitive (85% versus 79%) and to yield an earlier positive result (36 days versus 15 days) than for serum. However, DNA extraction from serum specimens is easier and faster than that from whole-blood specimens, and it allows the same specimen to be used for both galactomannan and PCR assays. In conclusion, the appropriate sample type for DNA extraction should be determined by the local requirements and the technical platforms available at each individual center. A combination of biomarker tests offered the best diagnostic utility for detecting IA.
Tárgyszavak:	Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban DIAGNOSING INVASIVE ASPERGILLOSIS REAL-TIME PCR FUNGAL-INFECTIONS FUMIGATUS DNA
Megjelenés:	Journal of Clinical Microbiology. - 51 : 5 (2013), p. 1445-1450. -
További szerzők:	Morton, C. Oliver Perry, Michael Heinz, Werner J. Paholcsek Melinda (1984-) (molekuláris biológus, genetikus) Alzheimer, Mona Rogers, T. R. Barnes, Rosemary A. Einsele, Hermann Loeffler, Juergen White, P. Lewis
Pályázati támogatás:	German Federal Ministry of Research and Education (BMBF) Egyéb Trans-European Cooperation ERA-NET PathoGenoMics Egyéb
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