Diagnosis of Cytomegalovirus in Bronchoalveolar Lavage by Polymerase Chain Reaction, in Comparison with Virus Isolation and Detection of Viral Antigen

Abstract

Bronchoalveolar lavage (BAL) products from 52 immunocompromised patients with symptoms of pulmonary infection was examined for cytomegalovirus (CMV) by virus isolation, polymerase chain reaction (PCR) and detection of CMV antigen by immunofluorescence or immunoperoxidase staining after short-term incubation in tissue culture and directly in BAL cells. We found that PCR detected all cases positive by virus isolation (15/52 samples) and the result was obtained within 5 h. PCR detected more cases of CMV than did virus isolation (22/52 samples). Positive PCR and negative virus isolation were consistent with probable CMV infection in 3/7 patients when other clinical and laboratory parameters of CMV infection were considered. The negative predictive value of PCR was high; none of 30 patients negative by PCR developed CMV pneumonia within the subsequent 2 months. Detection of CMV antigen after short-term incubation was rapid enough to be used in clinical practice, specific (100%) and with a sensitivity of 60%. Demonstration of CMV antigen in alveolar cells was highly specific (100%) but had too low a sensitivity (26.7%) to be used as the only rapid method. Our conclusion is that a combination of PCR and detection of CMV antigen after short-term incubation and directly in alveolar cells is optimal for rapid identification of CMV.

Detection of CMV Pneumonitis after Lung Transplantation Using PCR of DNA from Bronchoalveolar Lavage Cells

“…These patients showed no signs of CMV pneumonitis and developed no pneumonitis within 1 month, indicating its high negative predictive value for detecting CMV infections in the lung. Our data support similar findings by Eriksson et al [6]. The value of PCR in diagnosis is obvious considering our patient who died of CMV pneumonitis: no other diagnostic tool was successful in detecting CMV, except for PCR of CMV DNA in BAL cells.…”

“…Other investigators found this method valuable because of its high specificity of 100%. However, sensitivity in that study was only 26.7% [6]. Therefore, other complementary investigations are necessary to increase the overall sensitivity.…”

“…However, assays detecting antigenemia may not be reliable if patients are lymphocytopenic. Previous investigations demonstrated that PCR is a powerful tool for the diagnosis of CMV in immunocompromised patients [6, 13, 16, 17, 18, 19]. Lung-transplanted patients required additional considerations.…”

“…Cytospin slides were incubated with a mixture of mouse anti-CMV antibodies recognizing the DNA binding protein p52 and the IE1 68/72-kD protein (MO854, Dako, Hamburg, Germany) [6]at a dilution of 1:25 for 30 min. After washing with PBS, a second antibody (rabbit anti-mouse IgG, Dako, Z0259) and a third antibody (APAAP anti-mouse IgG, Dako, D0651) were applied, each at a dilution of 1:50 for 30 min.…”

Spectrum of Pulmonary Infections in Renal Transplant Recipients in the Tropics: A Single Center Study

“…BAL PCR for CMV was positive in 11 out of 22 episodes but only one case actually had CMV disease. This high positivity for CMV PCR in BAL was also obtained by Eriksson et al [29], who suggested that a positive CMV PCR in BAL cannot always be related to CMV disease and should be correlated with other clinical evidence of CMV disease. They suggested that a combination of PCR and detection of CMV antigen directly in alveolar cells is optimal for rapid and accurate identification of true CMV disease.…”