Use of real-time PCR on blood samples for diagnosis of invasive aspergillosis.
We developed a new quantitative system for diagnosis of invasive pulmonary aspergillosis (IPA) using real-time automated polymerase chain reaction (PCR). Intra-assay and interassay precision rates for in vitro examination were 2.53% and 2.20%, respectively, and the linearity of this assay was obtained when there were >20 copies/well. We examined 323 samples taken from 122 patients with hematological malignancies, including 33 patients with IPA and 89 control patients. Blood samples were subjected to PCR antigen detection methods, using enzyme-linked immunosorbent assay (ELISA) and determination of plasma (1-->3)-beta-D-glucan (BDG) concentration. The sensitivities of PCR, ELISA, and BDG measurement for diagnosis of IPA were 79%, 58%, and 67%, respectively; the specificities were 92%, 97%, and 84%. Positive findings on PCR preceded those of computed tomography by -0.3+/-6.6 days, those of BDG measurement by 6.5+/-4.9 days, and those of ELISA by 2.8+/-4.1 days. Real-time PCR was sensitive for IPA diagnosis, and quantitation was accurate.
Kami
M
M
Department of Hematology and Oncology, Graduate School of Medicine, University of Tokyo, Japan. kami-tky@umin.ac.jp
Fukui
T
T
Ogawa
S
S
Kazuyama
Y
Y
Machida
U
U
Tanaka
Y
Y
Kanda
Y
Y
Kashima
T
T
Yamazaki
Y
Y
Hamaki
T
T
Mori
S
S
Akiyama
H
H
Mutou
Y
Y
Sakamaki
H
H
Osumi
K
K
Kimura
S
S
Hirai
H
H
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001228:Aspergillosis / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology / Q000503:physiopathology; D001230:Aspergillus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005334:Fever; D005936:Glucans / Q000097:blood; D006801:Humans; D008172:Lung Diseases, Fungal / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology / Q000503:physiopathology; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014057:Tomography, X-Ray Computed / Q000379:methods; D047071:beta-Glucans
D005936:Glucans; D047071:beta-Glucans; C033363:beta-1,3-glucan