title: Automated detection of immunofluorescently labeled cytomegalovirus-infected cells in isolated peripheral blood leukocytes using decision tree analysis.
abstract: BACKGROUND
  Cytomegalovirus (CMV) infection continues to be a major problem for immunocompromised patients. Detection of viral antigens in leukocytes (antigenemia assay) is widely used for the diagnosis of CMV infection and for guiding antiviral therapy. The antigenemia technique, contingent upon the manual microscopic analysis of rare cells, is a laborious task that is subject to human error. In this study, we combine automated microscopy with artificial intelligence for reliable detection of fluorescently labeled CMV-infected cells.
  
  
  METHODS
  Cytospin preparations of peripheral blood leukocytes were immunofluorescently labeled for the CMV lower matrix phosphoprotein (pp65) and scanned in the Rare Event Imaging System (REIS), a fully automated image cytometer. The REIS detected potential positive objects and digitally recorded 49 measured cellular features for each identified case. The measurement data of these objects were analyzed by the See5 decision tree (DT) algorithm to ascertain whether they were true-positive detections.
  
  
  RESULTS
  The DT was built from the measurement data of 2,047 true- and 2,028 false-positive detections, collected from 32 patient samples. By designating misclassifications of false-negatives three times more costly, the 10-fold cross-validation sensitivity, specificity, and misclassification error of the assay was 94.3%, 56.2%, and 25%, respectively. The method was also validated using an independent test set of 21 patient samples, in which similar results were obtained.
  
  
  CONCLUSIONS
  To our knowledge, this study represents the first attempt to improve the accuracy of rare event image cytometry through the implementation of artificial intelligence methodology. Results suggest that cost-sensitive decision tree analysis of digitally measured cellular features vastly improves the performance of rare event image cytometry.
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA. andras_ladanyi@dfci.harvard.edu
authors.forename: Andras
authors.initials: A
authors.lastname: Ladanyi
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Sher
authors.forename: Anna
authors.initials: A
authors.lastname: Herlitz
authors.forename: Danielle E
authors.initials: DE
authors.lastname: Bergsrud
authors.forename: Stine-Kathrein
authors.initials: SK
authors.lastname: Kraeft
authors.forename: Jane
authors.initials: J
authors.lastname: Kepros
authors.forename: Gizelda
authors.initials: G
authors.lastname: McDaid
authors.forename: David
authors.initials: D
authors.lastname: Ferguson
authors.forename: Marie L
authors.initials: ML
authors.lastname: Landry
authors.forename: Lan Bo
authors.initials: LB
authors.lastname: Chen
mesh_terms: D001185:Artificial Intelligence; D001331:Automation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000302:isolation & purification*; D003663:Decision Trees; D019084:Fluorescent Antibody Technique, Indirect / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007962:Leukocytes / Q000821:virology*; D008856:Microscopy, Fluorescence; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity