Article

Detection of Aspergillus DNA by a nested PCR assay is able to improve the diagnosis of invasive aspergillosis in paediatric patients.

III. Medizinische Universitätsklinik, Medizinische Fakultät Mannheim, Universität Heidelberg, D-68167 Mannheim, Germany.
Journal of Medical Microbiology (impact factor: 2.5). 07/2009; 58(Pt 10):1291-7. DOI:10.1099/jmm.0.007393-0 pp.1291-7
Source: PubMed

ABSTRACT Fungal infections are a leading cause of morbidity and mortality in severely immunocompromised patients and have been increasing in incidence in recent years. Invasive aspergillosis (IA) is the most common filamentous fungal infection and is, in adults as well as in children, difficult to diagnose. Several PCR assays to detect Aspergillus DNA have been established, but so far, studies on molecular tools for the diagnosis of IA in children are few. We evaluated the results of a nested PCR assay to detect Aspergillus DNA in clinical samples from paediatric and adolescent patients with suspected IA. Blood and non-blood samples from immunocompromised paediatric and adolescent patients with suspected invasive fungal infection were sent for processing Aspergillus PCR to our laboratory. PCR results from consecutive patients from three university children's hospitals investigated between November 2000 and January 2007 were evaluated. Fungal infections were classified according to the EORTC classification on the grounds of clinical findings, microbiology and radio-imaging results. Two hundred and ninety-one samples from 71 patients were investigated for the presence of Aspergillus DNA by our previously described nested PCR assay. Two, 3 and 34 patients had proven, probable and possible IA, respectively. Sensitivity (calculated from proven and probable patients, n=5) and specificity (calculated from patients without IA, n=32) rates of the PCR assay were 80 and 81 %, respectively. Our nested PCR assay was able to detect Aspergillus DNA in blood, cerebrospinal fluid and bronchoalveolar lavage samples from paediatric and adolescent patients with IA with high sensitivity and specificity rates.

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    Article: Diagnostic potential of nested PCR, galactomannan EIA, and Beta-D-glucan for invasive aspergillosis in pediatric patients
    Parisa Badiee, Abdolvahab Alborzi, Mahammad Karimi, Bahman Pourabbas, Pedram Haddadi, Jalal Mardaneh, Mahsa Moieni
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    ABSTRACT: Introduction: Limited specific data and investigations are available for invasive aspergillosis (IA) in pediatric patients. We evaluated the diagnostic potential of three noninvasive tests including the Platelia Aspergillus EIA kit for using galactomannan antigen, (1,3)–β–D–glucan Detection Reagent Kit, and nested-PCR for Aspergillus DNA in sera. We evaluated the diagnostic potential of three noninvasive tests including EIA for galactomannan antigen (Platelia Aspergillus), nested PCR assay for Aspergillus DNA and test for (1→3)-β-D-glucan (Glucatell assay Kit). Methodology: All pediatric patients treated at the hematology/oncology unit who were at increased risk of developing invasive aspergillosis were enrolled. Clinical samples were examined for Aspergillus infections by mycological methods. Serial blood samples were collected twice weekly and evaluated by noninvasive tests. Results: We analyzed 230 consecutive blood samples from 62 pediatric patients. The incidence rate of invasive aspergillosis in the patients was found to be 27.4%, and the etiologic agents were Aspergillus flavus, Aspergillus fumigatus, and Aspergillus spp. The sensitivity, specificity, positive and negative predictive values, and likelihood ratios for positive and negative results of galactomannan in patients with proven and probable IA were 90%, 92%, 81.8%, 96%, 11.25, and 0.1; for beta–D–glucan they were 50%, 46%, 26%, 70.6%, 0.9, 0.9; and for nested-PCR they were 80%, 96.2%, 88.9%, 92.6%, 21, and 0.2, respectively. Conclusions: The conventional methods are not able to detect IA, due to the lack of valid and proper sampling. Galactomannan and nested-PCR tests in serum, with enough accuracy and reliability, can serve as noninvasive methods for the detection of IA in pediatric patients. However, the beta–D–glucan test cannot serve as an efficient diagnostic tool in those with hematologic disorders.
    The Journal of Infection in Developing Countries 04/2012; 6(4):352-357. · 1.19 Impact Factor
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Keywords

Aspergillus DNA
 
bronchoalveolar lavage samples
 
clinical samples
 
common filamentous fungal infection
 
described nested PCR assay
 
EORTC classification
 
Fungal infections
 
immunocompromised paediatric
 
invasive fungal infection
 
molecular tools
 
nested PCR assay
 
non-blood samples
 
PCR assay
 
PCR assays
 
PCR results
 
possible IA
 
processing Aspergillus PCR
 
radio-imaging results
 
specificity rates
 
university children's hospitals