Diagnosis of invasive fungal infections by a real-time panfungal PCR assay in pediatric patients undergoing intensive chemotherapy or allogeneic stem cell transplantion

2010 ◽  
Vol 222 (03) ◽  
Author(s):  
M Bernroitner ◽  
C Landlinger ◽  
L Baskova ◽  
S Preuner ◽  
M Van Grotel ◽  
...  
Keyword(s):  
Stem Cell ◽  
Real Time ◽  
Pediatric Patients ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Intensive Chemotherapy ◽  
Pcr Assay ◽  
Panfungal Pcr

Diagnosis of Invasive Fungal Infections by a Real-Time Panfungal PCR Assay in Pediatric Patients Undergoing Intensive Chemotherapy or Allogeneic Stem Cell Transplantion.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 343-343
Author(s):  
Christine Landlinger ◽  
Lenka Baskova ◽  
Sandra Preuner ◽  
Martine Van Grotel ◽  
Nico G. Hartwig ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Predictive Value ◽  
Pediatric Patients ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Immunocompromised Patients ◽  
Pcr Assay ◽  
Highly Sensitive ◽  
Panfungal Pcr

Abstract Abstract 343 Invasive fungal infections are life threatening events in severely immunocompromised patients, and there is urgent need for reliable screening methods facilitating rapid and broad detection of pathogenic fungi. We have established a two-reaction real-time PCR assay permitting highly sensitive detection and quantitative monitoring of more than 80 fungal pathogens, covering a large spectrum of moulds, yeasts and Zygomycetes (European patent No. 06817468.9). To assess the clinical potential of the panfungal real-time PCR assay, more than 600 consecutive specimens from 126 pediatric patients carrying a high risk of invasive fungal infections were analyzed. The results revealed an excellent correlation between PCR positivity and the presence of proven, probable or possible fungal infection according to the criteria of the European Organization for Research and Treatment of Cancer (EORTC), indicating a sensitivity of the assay of 96% (95%CI: 81-99.3%). Hence, the negative predictive value of the panfungal PCR assay presented is very high, and our current data indicate that molecular screening of patients during febrile neutropenic episodes by the assay can help prevent unnecessary toxicity resulting from empirical antifungal treatment in individuals who may not be at risk of imminent fungal disease. The specificity of the assay in the test cohort of patients was in the range of 76% (95%CI:62-87%), and the observations indicate that rapid species identification may be required to assess the positive predictive value for impending fungus-related disease. The availabel data provide a basis for appropriately designed clinical studies addressing the full diagnostic potential of fungal screening by highly sensitive, broad- spectrum molecular assays in severely immunocompromised patients. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Diagnosis of invasive fungal infections by a real-time panfungal PCR assay in immunocompromised pediatric patients

Leukemia ◽  
2010 ◽  
Vol 24 (12) ◽  
pp. 2032-2038 ◽  
Author(s):  
C Landlinger ◽  
S Preuner ◽  
L Bašková ◽  
M van Grotel ◽  
N G Hartwig ◽  
...  
Keyword(s):  
Real Time ◽  
Pediatric Patients ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Pcr Assay ◽  
Panfungal Pcr

Diagnosis of invasive fungal infections using real-time PCR assay in paediatric acute leukaemia induction

Mycoses ◽  
2012 ◽  
Vol 55 (4) ◽  
pp. 372-379 ◽  
Author(s):  
Sushil Mandhaniya ◽  
Sobuhi Iqbal ◽  
Surender Kumar Sharawat ◽  
Immaculata Xess ◽  
Sameer Bakhshi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Acute Leukaemia ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Pcr Assay

scholarly journals New Panfungal Real-Time PCR Assay for Diagnosis of Invasive Fungal Infections

2016 ◽  
Vol 54 (12) ◽  
pp. 2910-2918 ◽  
Author(s):  
Clara Valero ◽  
Laura de la Cruz-Villar ◽  
Óscar Zaragoza ◽  
María José Buitrago
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Fungal Infections ◽  
Melting Curve ◽  
Fungal Species ◽  
Invasive Fungal Infections ◽  
Clinical Samples ◽  
Melting Curve Analysis ◽  
Pcr Assay ◽  
Suitable Alternative

The diagnosis of invasive fungal infections (IFIs) is usually based on the isolation of the fungus in culture and histopathological techniques. However, these methods have many limitations often delaying the definitive diagnosis. In recent years, molecular diagnostics methods have emerged as a suitable alternative for IFI diagnosis. When there is not a clear suspicion of the fungus involved in the IFI, panfungal real-time PCR assays have been used, allowing amplification of any fungal DNA. However, this approach requires subsequent amplicon sequencing to identify the fungal species involved, increasing response time. In this work, a new panfungal real-time PCR assay using the combination of an intercalating dye and sequence-specific probes was developed. After DNA amplification, a melting curve analysis was also performed. The technique was standardized by using 11 different fungal species and validated in 60 clinical samples from patients with proven and probable IFI. A melting curve database was constructed by collecting those melting curves obtained from fungal species included in the standardization assay. Results showed high reproducibility (coefficient of variation [CV] < 5%; r > 0.95) and specificity (100%). The overall sensitivity of the technique was 83.3%, with the group of fungi involved in the infection detected in 77.8% of the positive samples with IFIs covered by molecular beacon probes. Moreover, sequencing was avoided in 67.8% of these “probe-positive” results, enabling report of a positive result in 24 h. This technique is fast, sensitive, and specific and promises to be useful for improving early diagnosis of IFIs.

scholarly journals Clinical evaluation of an in-house panfungal real-time PCR assay for the detection of fungal pathogens

Infection ◽  
2020 ◽  
Vol 48 (3) ◽  
pp. 345-355 ◽  
Author(s):  
Iris Camp ◽  
Gabriele Manhart ◽  
Claudia Schabereiter-Gurtner ◽  
Kathrin Spettel ◽  
Brigitte Selitsch ◽  
...  
Keyword(s):  
Clinical Evaluation ◽  
Fungal Pathogens ◽  
Fungal Infections ◽  
Wide Spectrum ◽  
Invasive Fungal Infections ◽  
Diagnostic Tools ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Blood Samples ◽  
Panfungal Pcr

Abstract Purpose Due to an increasing incidence of invasive fungal infections, the availability of reliable diagnostic tools for the fast detection of a wide spectrum of fungal pathogens is of vital importance. In this study, we aimed to conduct an extensive clinical evaluation of a recently published in-house panfungal PCR assay on samples from suspected invasive fungal infections. Methods Overall 265 clinical samples from 232 patients with suspected invasive fungal disease (96 deep airway samples, 60 sterile fluids, 50 tissue biopsies, and 59 blood samples) were included. All samples underwent standard culture-based diagnostics and were additionally analyzed with our panfungal PCR assay. Results Overall, 55.1% of agreement between culture and the panfungal PCR was observed; in 17% of all samples partial concordance was noted, while results between culture and our PCR assay were not in agreement in 27.9%. Our panfungal assay performed better in samples from normally sterile sites, while samples from the deep airways yielded the highest rate of discordant (39.6%) results. In two tissue and three blood samples an invasive pathogen was only detected by PCR while cultures remained negative. Conclusion In combination with routine methods, our panfungal PCR assay is a valuable diagnostic tool. Patients at risk for invasive fungal infections might profit from the reduced time to pathogen identification.

scholarly journals Cost Analysis of the Use of Voriconazole, Posaconazole and Micafungin in the Primary Prophylaxis of Invasive Fungal Infections in Recipients of Allogeneic Hematopoietic Stem Cell Transplants

10.36469/9832 ◽  
2015 ◽  
Vol 3 (2) ◽  
pp. 153-161
Author(s):  
Santiago Grau ◽  
Carlos Solano ◽  
Carol García-Vidal ◽  
Isidro Jarque ◽  
Jon A. Barrueta ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cost Analysis ◽  
Hematopoietic Stem Cell ◽  
Fungal Infections ◽  
Primary Prophylaxis ◽  
Antifungal Prophylaxis ◽  
Invasive Fungal Infections ◽  
Efficacy Rate ◽  
Hematopoietic Stem ◽  
The Cost

Objectives: Compare the cost of the primary prophylaxis of invasive fungal infections (IFI) with voriconazole, posaconazole, and micafungin in patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) in hospitals of the National Health System (NHS) in Spain. Methods: A cost analysis was made for 100 days and 180 days of prophylaxis and a decision tree model was developed. The efficacy rate of IFI prophylaxis and survival rate with liposomal amphotericin B treatment of prophylaxis failures were obtained from randomized trials and a meta-analysis of mixed treatment comparisons. The model simulation was interrupted with IFI treatment (prophylaxis failures). The costs of medication and its intravenous administration in the hospital (in the case of micafungin) were considered. Results: In the non-modeled analysis, the savings per patient of prophylaxis with voriconazole ranged from €1,709 to €9,655 compared with posaconazole oral solution, from €1,811 to €9,767 compared with posaconazole gastro-resistant tablets and from €3,376 to €7,713 compared with micafungin. In the modeled analysis, the mean cost per patient of the prophylaxis and treatment of IFIs was €6,987 to €7,619 with voriconazole, €7,749 with posaconazole, and €22,424 with micafungin. Therefore, the savings per patient of prophylaxis with voriconazole was €130 to €3,664 and €11,132 to €30,374 compared with posaconazole and micafungin, respectively. The result remained stable after modification of the number of days of antifungal prophylaxis and the cost of antifungal treatment of failures. Conclusion: Taking into account this model, antifungal prophylaxis with voriconazole in recipients of hematopoietic progenitor transplants, compared with posaconazole or micafungin, may represent savings for hospitals in Spain.

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