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Published on: March 22, 2012
Aspergillus PCR testing: results from a prospective PCR study within the AmBiLoad trial
Margit Hummel1, Birgit Spiess, Oliver A Cornely
1III Medizinische Klinik, Klinikum Mannheim, University of Heidelberg, Mannheim, Germany. margithummel@yahoo.de
Objectives:
Invasive fungal infection (IFI) is a major cause of morbidity and mortality in severely immunocompromised patients and is difficult to diagnose. The significance of molecular methods for diagnosis of IFI is still controversial. In a subset of patients treated within the AmBiLoad Trial, samples were investigated prospectively by a nested Aspergillus PCR assay to re-evaluate the significance of PCR in this setting.
Patients And Methods:
In the randomized, prospective multicenter AmBiLoad trial, patients with proven or probable IFI were randomized to receive liposomal amphotericin B (L-AMB) 3 or 10 mg/kg QD for 14 d followed by L-AMB 3 mg/kg QD. From 91 patients, 459 serial samples (98% blood samples) were investigated by a nested PCR assay for Aspergillus DNA. All samples were investigated in our laboratory with a previously described nested and a quantitative PCR assay. As required by the study protocol, serial Aspergillus antigen galactomannan was performed. IFI was defined according to modified EORTC/MSG 2002 criteria as applied in the AmBiLoad trial.
Results:
Seven and 52 patients had proven and probable IFI according to modified EORTC/MSG criteria, respectively. The median number of samples investigated per patient was 4. Seventy percent of samples were obtained during treatment with antifungal study medication. Forty-three samples gave positive PCR results. Patients with an unfavorable outcome had a significantly higher rate of positive PCR results (48% versus 21%).
Conclusions:
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.
Insights
Molecular Aspergillus PCR testing shows limited sensitivity during antifungal therapy. Persistently positive PCR results may indicate a poor prognosis in immunocompromised patients, warranting further investigation.
Area of Science:
- Medical Mycology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Invasive fungal infections (IFI) pose significant risks for immunocompromised patients.
- Accurate and timely diagnosis of IFI is challenging.
- The role of molecular methods like PCR for IFI diagnosis remains debated.
Purpose of the Study:
- To prospectively evaluate the significance of a nested Aspergillus PCR assay in diagnosing IFI.
- To re-assess the utility of PCR in a subset of patients from the AmBiLoad Trial.
Main Methods:
- A nested PCR assay was used to detect Aspergillus DNA in serial samples from 91 patients with proven or probable IFI.
- Samples were collected during the AmBiLoad Trial, with 70% obtained during antifungal treatment.
- Aspergillus antigen galactomannan was also monitored as per study protocol.
Main Results:
- Of 459 samples, 43 tested positive by PCR.
- Patients with unfavorable outcomes showed a significantly higher rate of positive PCR results (48%) compared to those with favorable outcomes (21%).
Conclusions:
- The sensitivity of Aspergillus PCR is constrained when patients are undergoing antifungal therapy.
- A trend suggests that persistent positive PCR results correlate with a poor prognosis, requiring further validation.
