Diagnosis of cytomegalovirus pneumonia by quantitative polymerase chain reaction using bronchial washing fluid from patients with hematologic malignancies

Oncotarget. 2017 Jun 13;8(24):39736-39745. doi: 10.18632/oncotarget.14504.

Abstract

Background: The incidence of cytomegalovirus (CMV) pneumonia is increasing in patients diagnosed with hematologic malignancies. The utility of CMV-DNA viral load measurement has not been standardized, and viral cut-off values have not been established. This study was designed to investigate the utility of CMV quantitative real-time PCR (qRT-PCR) using bronchial washing fluid.

Methods: We retrospectively reviewed the microbiologic and pathologic results of bronchial washing fluid and biopsy specimens in addition to the patients' clinical characteristics.

Results: A total of 565 CMV qRT-PCR assays were performed using bronchial washing fluid from patients with hematologic malignancies. Among them, 101 were positive for CMV by qRT-PCR; of these, 24 were diagnosed with CMV pneumonia and 70 with CMV infection, and 7 were excluded due to a diagnosis of invasive pulmonary aspergillosis rather than viral pneumonia. The median CMV load determined by qPCR was 1.8 × 105 copies/mL (3.6 103-1.5 × 108) in CMV pneumonia patients and 3.0 × 103 copies/mL (5.0 × 102-1.1 × 105) in those diagnosed with CMV infection (P < 0.01). Using the ROC curve, the optimal inflection points were 18,900 copies/mL (137,970 IU/mL) in post-bone marrow transplantation (BMT) patients, 316,415 copies/mL (2,309,825 IU/mL) in no-BMT patients and 28,774 copies/mL (210,054 IU/mL) in all patients.

Conclusions: The CMV titers in bronchial washing fluid determined by qRT-PCR differed significantly between patients diagnosed with CMV pneumonia and those with CMV infection. The viral cut-off values in bronchial washing fluid were suggested for the diagnosis of CMV pneumonia, which were different depending on the BMT status.

Keywords: cytomegalovirus; hematologic neoplasms; pneumonia; real-time polymerase chain reaction.

MeSH terms

  • Bronchoalveolar Lavage Fluid / chemistry*
  • Cytomegalovirus / genetics*
  • Cytomegalovirus / isolation & purification
  • Cytomegalovirus Infections / diagnosis*
  • Cytomegalovirus Infections / genetics
  • Cytomegalovirus Infections / virology
  • DNA, Viral / genetics
  • Female
  • Follow-Up Studies
  • Hematologic Neoplasms / complications*
  • Humans
  • Male
  • Middle Aged
  • Pneumonia / diagnosis*
  • Pneumonia / genetics
  • Pneumonia / virology
  • Prognosis
  • ROC Curve
  • Real-Time Polymerase Chain Reaction / methods*
  • Retrospective Studies
  • Viral Load

Substances

  • DNA, Viral