Detection of cytomegalovirus (CMV) in granulocytes by polymerase chain reaction compared with the CMV antigen test

Abstract

To compare the sensitivity and suitability of detection of active cytomegalovirus (CMV) infection by using monoclonal antibodies against CMV antigen (antigen test to detect antigenemia) and the polymerase chain reaction (PCR; to detect viral DNA) in granulocytes, 19 heart and 2 lung transplant recipients were closely monitored by these tests for at least 3 months after transplantation. All patients were CMV seropositive or had a seropositive donor. In total, 201 samples were tested; 46 were positive by both tests, 9 samples showed only antigenemia, 54 samples were positive by PCR only, and 102 samples were negative by both tests. PCR was positive earlier after transplantation in eight patients, whereas antigenemia was positive earlier after transplantation in one patient. In another four patients, both tests were positive at the same time. PCR was, on average, positive for a longer period of time. Discordant results showing a positive antigen test and a negative PCR were partly due to sampling error; some were positive by PCR on retesting. Samples which were negative by the antigen test and positive by PCR were taken at the beginning or at the end of an active CMV infection. In two patients, no active CMV infection was detected by the antigen test, cultures of urine and saliva, or serology, although PCR was positive for a long period of time in the two patients.

Monitoring for cytomegalovirus infection in organ transplant recipients: Analysis of pp65 antigen, DNA and late mRNA in peripheral blood leukocytes

“…It is essential for a timely application of antiviral therapy to prevent the life-threatening consequences of viral disease and also to detect the efficacy of the treatment. For this purpose, the demonstration of viral antigens in peripheral blood leukocytes (PMNLs) and of viral nucleic acids in the same cells or in sera has proven to be of valid support Boland et al, 1992;Cunningham et al, 1995].…”

Low correlation of human cytomegalovirus DNA amplification by polymerase chain reaction with cytomegalovirus disease in organ transplant recipients

“…The shell vial culture using immunoperoxidase or immunofluorescence staining of infected fibroblasts enables virus isolation from peripheral blood leukocytes (PBL) and various body fluids as early as 24-36 hours post-inoculation and has been shown to be a more reliable marker for active infection than serology [Schacherer et al, 1988;Weber et al, 1992b1. More recently, the quantitative pp65 structural antigen detection in PBL with the alkaline phosphatase anti-alkaline phosphatase (APAAP) or indirect immunofluorescence staining has been shown to be an easy and rapid test for early diagnosis of HCMV infection and correlates well with viraemia [Bein et al, 1991;Gerna et al, 1991a,bl. The detection of HCMV encoded DNA sequences by the polymerase chain reaction (PCR) in PBL and c h i - cal samples offers a theoretical sensitivity of one molecule containing the target DNA sequence in 10 pg foreign DNA [Kuhn et al, 19911. Recent studies using PCR to detect HCMV DNA in clinical samples showed promising results [Boland et al, 1992;Buffone et al, 1990;Einsele et al, 1991;Fenner et al, 1991;Olive et al, 1989;Zipeto et al, 19921. Nevertheless, as a consequence of its extreme sensitivity, the predictive value of PCR for the diagnosis of cytomegalovirus disease remains controversial [Delgado et al, 19921.…”

Detection improvement of cytomegalovirus antigen in human peripheral blood using monoclonal antibodies and automated reading of cell preparations

“…Previous studies have shown that the detection of cytomegalovirus antigenaemia, which is the presence of CMV antigen in peripheral blood leucocytes, is a sensitive marker for the detection of an active CMV infection [2,3,[5][6][7][8]10,11,13]. In this study, we have optimized the detection of antigenaemia in two ways.…”