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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:	BIBFORM059207
Első szerző:	Paholcsek Melinda (molekuláris biológus, genetikus)
Cím:	Combining standard clinical methods with PCR showed improved diagnosis of invasive pulmonary aspergillosis in patients with hematological malignancies and prolonged neutropenia / Melinda Paholcsek, Gabor Fidler, Jozsef Konya, Laszlo Rejto, Gabor Mehes, Evelin Bukta, Juergen Loeffler, Sandor Biro
Dátum:	2015
ISSN:	1471-2334
Megjegyzések:	Background: We assessed the diagnostic value of standard clinical methods and combined biomarker testing(galactomannan assay and polymerase chain reaction screening) in a prospective case?control study to detectinvasive pulmonary aspergillosis in patients with hematological malignancies and prolonged neutropenia.Methods: In this observational study 162 biomarker analyses were performed on samples from 27 febrile neutropenicepisodes. Sera were successively screened for galactomannan antigen and for Aspergillus fumigatus specific nucleic acidtargets. Furthermore thoracic computed tomography scanning was performed along with bronchoscopy with lavagewhen clinically indicated. Patients were retrospectively stratified to define a case-group with "proven" or "probable"invasive pulmonary aspergillosis (25.93 %) and a control-group of patients with no evidence for of invasive pulmonaryaspergillosis (74.07 %). In 44.44 % of episodes fever ceased in response to antibiotic treatment (group II). Empiricalantifungal therapy was administered for episodes with persistent or relapsing fever (group I). 48.15 % of patientsdied during the study period. Postmortem histology was pursued in 53.85 % of fatalities.Results: Concordant negative galactomannan and computed tomography supported by a polymerase chainreaction assay were shown to have the highest discriminatory power to exclude invasive pulmonary aspergillosis.Bronchoalveolar lavage was performed in 6 cases of invasive pulmonary aspergillosis and in 15 controls. Althoughbronchoalveolar lavage proved negative in 93 % of controls it did not detect IPA in 86 % of the cases. Remarkablypost mortem histology convincingly supported the presence of Aspergillus hyphae in lung tissue from a single casewhich had consecutive positive polymerase chain reaction assay results but was misdiagnosed by both computedtomography and consistently negative galactomannan assay results. For the galactomannan enzyme-immunoassay thediagnostic odds ratio was 15.33 and for the polymerase chain reaction assay it was 28.67. According to Cohen's kappaour in-house polymerase chain reaction method showed a fair agreement with the galactomannan immunoassay.Combined analysis of the results from the Aspergillus galactomannan enzyme immunoassay together with thosegenerated by our polymerase chain reaction assay led to no misdiagnoses in the control group.Conclusion: The data from this pilot-study demonstrate that the consideration of standard clinical methods combinedwith biomarker testing improves the capacity to make early and more accurate diagnostic decisions.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Invasive pulmonary aspergillosis Biomarkers Combination testing Acute leukemia Neutropenic fever
Megjelenés:	BMC Infectious Diseases. - 15 : 1 (2015), p. 251. -
További szerzők:	Fidler Gábor (1987-) (molekuláris biológus, genetikus) Kónya József (1964-) (szakorvos, klinikai mikrobiológus) Rejtő László (1963-) (belgyógyász, haematológus) Méhes Gábor (1966-) (patológus) Bukta Evelin Loeffler, Juergen Biró Sándor (1949-) (molekuláris genetikus)
Pályázati támogatás:	SROP-4.2.2.B-15/1/KONV-2015-0001 Egyéb TÁMOP-4.2.4.A/ 2-11/1-2012-0001 TÁMOP
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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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001-es BibID:	BIBFORM083399
Első szerző:	White, P. Lewis
Cím:	Comment on: T2Candida MR as a predictor of outcome in patients with suspected invasive candidiasis starting empirical antifungal treatment : a prospective pilot study / P. Lewis White, Rosemary A. Barnes, Rebecca Gorton, Mario Cruciani, Juergen Loeffler, Fungal PCR Initiative
Dátum:	2019
Megjegyzések:	We read with interest the article of Munoz ~ et al.1 describing the performance of the T2Candida MR assay in patients receiving empirical antifungal therapy for suspected invasive candidiasis (IC).1 The study was well designed and fully warranted in attempting to address the demands of managing patients at risk of fungal disease and provides much needed real-life data on the performance of the T2 platform. However, unlike the authors, we feel the performance of the T2 assay in this study was insufficient to discontinue antifungal therapy.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok hozzászólás folyóiratcikk
Megjelenés:	The Journal of Antimicrobial Chemotherapy. - 74 (2019), p. 532-533. -
További szerzők:	Barnes, Rosemary A. Gorton, Rebecca Cruciani, Mario Loeffler, Juergen Paholcsek Melinda (1984-) (molekuláris biológus, genetikus) Fungal PCR Initiative
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