Use of Real-Time PCR on Blood Samples for Diagnosis of Invasive Aspergillosis

Clin Infect Dis. 2001 Nov 1;33(9):1504-12. doi: 10.1086/323337. Epub 2001 Oct 4.

Abstract

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.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Aspergillosis / blood
  • Aspergillosis / diagnosis*
  • Aspergillosis / microbiology
  • Aspergillosis / physiopathology
  • Aspergillus / genetics
  • Aspergillus / immunology
  • Aspergillus / isolation & purification
  • Aspergillus / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Fever
  • Glucans / blood
  • Humans
  • Lung Diseases, Fungal / blood
  • Lung Diseases, Fungal / diagnosis*
  • Lung Diseases, Fungal / immunology
  • Lung Diseases, Fungal / microbiology
  • Lung Diseases, Fungal / physiopathology
  • Male
  • Middle Aged
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Time Factors
  • Tomography, X-Ray Computed / methods
  • beta-Glucans*

Substances

  • Glucans
  • beta-Glucans
  • beta-1,3-glucan