Diagnosis of invasive mold infection by real-time quantitative PCR.
We report the design and evaluation of a quantitative real-time polymerase chain reaction (PCR) assay to diagnose invasive mold infection (IMI) by detecting mold DNA in the serum. This assay detected 200 fg to 20 ng (5-log range) mold DNA and permitted a cutoff of 110 fg (3 genomes). Human or candidal DNA was not amplified. Specificity also was demonstrated by negative results in all 35 patients (76 serum samples) with unlikely IMI at the cutoff. For patients with possible, probable, and documented IMI diagnosed by a combination of clinical, microbiologic, and histologic criteria, this real-time PCR showed positivity in 40% (12/30), 68% (19/28), and 85% (11/13) cases, respectively, in testing of multiple serum samples. The overall serum positivity rate for these patients was 15.1% (73/483). Quantitative analysis of the positive serum samples estimated the bodily circulating mold burden to be 1.6 x 10(5) genomes (5.3 ng) by geometric mean with 4.2 x 10(7) genomes (1,400 ng) the highest. These results suggest that for the diagnosis of IMI, this real-time PCR may be a promising alternative to other invasive methods. Further evaluation is underway.
Pham
Audrey S
AS
Laboratory Medicine, University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX 77030, USA.
Tarrand
Jeffrey J
JJ
May
Gregory S
GS
Lee
Ming S
MS
Kontoyiannis
Dimitrios P
DP
Han
Xiang Y
XY
D001228:Aspergillosis / Q000097:blood / Q000175:diagnosis*; D001230:Aspergillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000737:chemistry; D004271:DNA, Fungal / Q000097:blood; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004271:DNA, Fungal