<i>Aspergillus</i> PCR testing: results from a prospective PCR study within the AmBiLoad trial

Hummel, Margit; Spieß, Birgit; Cornely, Oliver A.; Dittmer, Martin; Mörz, Handan; Buchheidt, Dieter

doi:10.1111/j.1600-0609.2010.01452.x

Cited by 37 publications

(23 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Lower sensitivity rates of PCR with blood samples during antifungal treatment have also been reported in both previous and recent studies (18,24), but at the current time, the impact of antifungal therapy in terms of the recovery of Aspergillus and related markers in the lung remains to be clearly defined (17). Falsenegative results were documented in only two of our patients.…”

Section: Discussionsupporting

confidence: 76%

“…Rationally, detection of fungal markers in the lungs (i.e., at the site of the disease) might be more sensitive than that in the serum (4), and in particular, serum GM levels may be lower in patients with airway IA than in those with angioinvasive aspergillosis (16). Also, Aspergillus DNAemia is probably only transient, and the most suitable time point for blood sampling for PCR has yet to be defined (18).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Diagnosis of Invasive Aspergillosis by a Commercial Real-Time PCR Assay for Aspergillus DNA in Bronchoalveolar Lavage Fluid Samples from High-Risk Patients Compared to a Galactomannan Enzyme Immunoassay

Torelli¹,

Sanguinetti²,

Moody

et al. 2011

J Clin Microbiol

120

View full text Add to dashboard Cite

Culture-independent molecular techniques such as real-time PCRs offer the potential for early diagnosis of invasive aspergillosis (IA), thereby reducing the disease-associated mortality rate. PCR-based testing is presently excluded from disease-defining consensus criteria due to lack of standardization and clinical validation. A single-center prospective study was conducted to investigate the performance of the commercially available MycAssay Aspergillus test for detecting Aspergillus DNA in patients with suspicion of IA. To this end, a total of 158 bronchoalveolar lavage (BAL) fluid specimens that were consecutively collected from hematology (n ‫؍‬ 68) and intensive care unit (n ‫؍‬ 90) patients were examined. Sixteen of 17 (94.1%) specimens from patients with proven/probable IA were MycAssay positive, and 15 of these 16 patients were also positive by an "in-house" PCR assay. A total of 139 of 141 (98.6%) specimens from patients without proven/probable IA were MycAssay negative. Fifteen of 16 (94.1%) MycAssay-positive patients were also positive for BAL fluid galactomannan (GM) at an index cutoff of >1.0 (index range, 1.1 to 8.3), as were 3 patients without IA but with pulmonary fusariosis. Interestingly, in seven of the PCR-positive BAL specimens that tested culture positive for Aspergillus species, cycle threshold values were earlier than those of specimens with a culture-negative result. In conclusion, the MycAssay Aspergillus PCR appears to be a sensitive and specific molecular test for the diagnosis of IA, and its performance is comparable to that of the GM assay. However, more large studies are necessary to firmly establish its clinical utility in high-risk settings.

show abstract

Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Diagnosis of Invasive Aspergillosis by a Commercial Real-Time PCR Assay for Aspergillus DNA in Bronchoalveolar Lavage Fluid Samples from High-Risk Patients Compared to a Galactomannan Enzyme Immunoassay

Torelli¹,

Sanguinetti²,

Moody

et al. 2011

J Clin Microbiol

120

View full text Add to dashboard Cite

show abstract

“…As reported in other studies, our understanding of DNA release and kinetics in fungal infections is still poor, and Aspergillus DNAemia is likely to be only transient (12,13).…”

Section: Discussionmentioning

confidence: 82%

Prospective Evaluation of Clinical and Biological Markers To Predict the Outcome of Invasive Pulmonary Aspergillosis in Hematological Patients

et al. 2012

View full text Add to dashboard Cite

Early evaluation of treatment efficacy in invasive aspergillosis (IA), a leading cause of morbidity and mortality in hematological patients, remains a challenge. We conducted a prospective study to evaluate the performance of different markers in predicting the outcome of patients with IA. Both clinical and biological criteria were assessed 7, 14, 21, and 45 days after inclusion in the study, and mortality was assessed at day 60. The association between baseline data and their evolution and the day 45 response to treatment was analyzed. A total of 57 patients (4 with proven, 44 with probable, and 9 with possible aspergillosis according to the revised EORTC/MSG [European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and National Institute of Allergy and Infectious Diseases Mycoses Study Group] definitions) were included. At day 45, 30 patients (53%) were determined to be responders, 25 (44%) were nonresponders, and 2 were not able to be evaluated. Twenty patients died within the 60 days of follow-up. We found that a poor day 45 outcome was associated with patients who had high baseline serum galactomannan (GM) antigen levels and those receiving steroids at the time of IA. A consistently negative serum GM index was associated with a good outcome, and the day 14 clinical evaluation was predictive of the day 45 outcome. No association was found between Aspergillus antibodies or DNA detection and patients' outcome. We conclude that the GM index value at diagnosis of IA, GM index kinetics, and clinical evaluation at day 14 are good markers for predicting the outcome of patients with IA and should be taken into account for adapting antifungal treatment.

show abstract

“…One could assume that antifungal treatment, by causing generalized mycelium breakdown, would lead to increased yields of the PCR assays, at least transiently, before fungal loads are significantly decreased. However, this hypothesis is undermined by reports from clinical studies and experimental models of IA, which show that PCR performance is negatively affected by antifungal treatment (132,141,143,177). One explanation for this paradox could be the fact that most DNA extraction protocols focus on methods of fungal cell lysis which can destroy free-floating fungal DNA, and thus render it undetectable (175).…”

Section: Pcrmentioning

confidence: 99%

“…Researchers studying these effects on a rat inhalation model of IA found that the measured reduction of PCR sensitivity with antifungal therapy could be attributed only to posaconazole and caspofungin, not to amphotericin B (179). However, other studies have failed to reach the same conclusion (141,177). Interestingly, a recent study evaluating the performance of PCR to diagnose IA by use of BAL fluid after at least one full daily dose of antifungal therapy showed that its performance was not significantly affected unless two or more antifungals were used to treat the patient (143).…”

Section: Pcrmentioning

confidence: 99%

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

et al. 2014

View full text Add to dashboard Cite

SUMMARY Invasive fungal infections constitute a serious threat to an ever-growing population of immunocompromised individuals and other individuals at risk. Traditional diagnostic methods, such as histopathology and culture, which are still considered the gold standards, have low sensitivity, which underscores the need for the development of new means of detecting fungal infectious agents. Indeed, novel serologic and molecular techniques have been developed and are currently under clinical evaluation. Tests like the galactomannan antigen test for aspergillosis and the β-glucan test for invasive Candida spp. and molds, as well as other antigen and antibody tests, for Cryptococcus spp., Pneumocystis spp., and dimorphic fungi, have already been established as important diagnostic approaches and are implemented in routine clinical practice. On the other hand, PCR and other molecular approaches, such as matrix-assisted laser desorption ionization (MALDI) and fluorescence in situ hybridization (FISH), have proved promising in clinical trials but still need to undergo standardization before their clinical use can become widespread. The purpose of this review is to highlight the different diagnostic approaches that are currently utilized or under development for invasive fungal infections and to identify their performance characteristics and the challenges associated with their use.

show abstract

Aspergillus PCR testing: results from a prospective PCR study within the AmBiLoad trial

Abstract: The sensitivity of Aspergillus PCR testing is limited during antifungal therapy. The tendency for persistently positive PCR results to indicate a poor prognosis has to be confirmed in further studies.

Cited by 37 publications

References 28 publications

Diagnosis of Invasive Aspergillosis by a Commercial Real-Time PCR Assay for Aspergillus DNA in Bronchoalveolar Lavage Fluid Samples from High-Risk Patients Compared to a Galactomannan Enzyme Immunoassay

Diagnosis of Invasive Aspergillosis by a Commercial Real-Time PCR Assay for Aspergillus DNA in Bronchoalveolar Lavage Fluid Samples from High-Risk Patients Compared to a Galactomannan Enzyme Immunoassay

Prospective Evaluation of Clinical and Biological Markers To Predict the Outcome of Invasive Pulmonary Aspergillosis in Hematological Patients

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

Contact Info

Product

Resources

About