Evaluation of PCR for Detection of DNA Specific for Aspergillus Species in Sera of Patients with Various Forms of Pulmonary Aspergillosis

Abstract

Pulmonary aspergillosis is classified into invasive, saprophytic, and allergic forms. In this study, we evaluated the usefulness of PCR for differentiating between different forms of aspergillosis or in monitoring disease activity during treatment by detecting DNA specific for Aspergillus species in the serum. Nested PCR was used to detect Aspergillus DNA in the sera of 30 patients with various forms of pulmonary aspergillosis. The results were compared with those of latex agglutination tests for detecting galactomannan antigen. We also examined the serial changes in the results of nested PCR during and after treatment of a subgroup of patients with invasive pulmonary aspergillosis with amphotericin B. The highest proportion of positive nested PCR results were in patients with invasive aspergillosis (10 of 12; 83%), while patients with pulmonary aspergilloma had the lowest frequency of positive tests (1 of 9; 11%). These results suggested that the sensitivity of the nested PCR depends on the extent of invasion by Aspergillus species. Serial assays showed that the results of nested PCR became negative shortly after commencement of antifungal treatment and that such changes did not correlate with clinical responsiveness to treatment. Our results indicate the potential usefulness of nested PCR with serum samples for the diagnosis of invasive aspergillosis and the detection of a shift in the status of infection from a noninvasive type to invasive aspergillosis. However, the results of the nested PCR did not correlate with the response to antifungal treatment.

The prospective evaluation of a nested polymerase chain reaction assay for the early detection of Aspergillus infection in patients with leukaemia or undergoing allograft treatment

“…It does appear that, as in case of patient O, the infection is detected one week and not the next. This phenomenon has been reported (Van Burik et al , 1998; Yamakami et al , 1998; Lass‐Flörl et al, 2001). The ability of the PCR to detect Aspergillus may have been affected by the presence of antifungal drugs.…”

Aspergillus‐Related Lung Disease

“…Tests are also available to detect aspergillus antibodies (120,121); however, these tests depend on a normal host immune response, which is absent in immunosupressed patients. A polymerase chain reaction test to detect DNA of Aspergillus species has been developed (122)(123)(124).…”

Clinical evaluation of a polymerase chain reaction assay to detect Aspergillus species in bronchoalveolar lavage samples of neutropenic patients

“…A number of previous studies evaluating PCR‐mediated detection of Aspergillus species showed significantly improved sensitivity but involved assays with different methods and objectives, partly to optimize culture assays, partly for typing in epidemiological studies (Aufauvre‐Brown et al , 1992; van Belkum et al , 1993; Girardin et al , 1994; Anderson et al , 1996; Rath et al , 1996; Brandt et al , 1998; Fletcher et al , 1998; Radford et al , 1998). Studies with clinical samples, such as blood or BAL, were mostly done retrospectively and, with few exceptions, with small numbers of patients (Spreadbury et al , 1993; Tang et al , 1993; Makimura et al , 1994; Melchers et al , 1994; Bretagne et al , 1995, 1998; Verweij et al , 1995a; Walsh et al , 1995; Kappe et al , 1996; Yamakami et al , 1996, 1998; Einsele et al , 1997, 1998; Jones et al , 1998; Van Burik et al , 1998; Kawamura et al , 1999; Skladny et al , 1999). Results from recent studies screening blood samples from high‐risk patients with PCR assays seem promising to identify a population at highest risk for invasive fungal disease (Einsele et al , 1998; Hebart et al , 2000a,b; Lass‐Flörl et al , 2001).…”