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

Abstract

Seventy-five organ transplant recipients underwent prolonged virological and serological follow-up for early detection of human cytomegalovirus (HCMV) infection after transplantation. HCMV DNA detection by nested polymerase chain reaction (PCR) and HCMV early structural antigen (pp65) detection were carried out in 576 peripheral blood leucocyte (PBL) samples. Furthermore, 563 blood specimens were investigated by a commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulins G, M, and A against HCMV structural antigens. In eight of nine symptomatic organ transplant recipients, HCMV DNA was detected in one or more consecutive blood samples. HCMV DNA PCR was also positive in one or more samples from eight patients who never developed HCMV-related symptoms. HCMV pp65 antigen was detected almost exclusively in PBL samples from organ transplant recipients suffering from HCMV disease. However, antigenaemia was not detected in four PCR positive patients presenting clinical signs attributable to HCMV infection. Two of the initially HCMV DNA positive samples were not confirmed by retesting and hybridisation. The results of the present study demonstrate that despite the high specificity of nested PCR, HCMV DNA may be detected in the absence of clinical symptoms attributable to HCMV infection. In asymptomatic reactivation, limited replication of viral DNA may be responsible for positive results of PCR without any clinical relevance. In this context, pp65-antigen detection from PBL seems to have a better prognostic value, but is not always detected when clinical symptoms are present.

Quantification of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients by Real-Time PCR

“…Furthermore, samples with low levels of antigenemia and a large number of copies of CMV DNA were observed. Similar differences between the results of an antigenemia assay and quantitative PCR have been reported by others (5,14,21), but the clinical significance of large CMV DNA copy numbers with a low or a negative antigenemia assay result remains to be elucidated.…”

Measurement of Human Cytomegalovirus Loads by Quantitative Real-Time PCR for Monitoring Clinical Intervention in Transplant Recipients

“…Therefore, qualitative PCR-based assays are probably too sensitive for clinical purposes. The plus-minus answer they supply is overly simplistic for clinical purposes, as it only allows for a single threshold to initiate therapy and provides limited feedback on response to therapy (21,28,35).…”

Quantification of Human Cytomegalovirus DNA by Real-Time PCR

“…Others have reported a similar difference between antigenemia and quantitative PCR results (10,16), but the clinical significance of a high HCMV DNA load associated with a low antigenemia remains to be established.…”

“…The detection of the pp65 antigen in leukocytes is a sensitive method widely used for the early diagnosis of HCMV infection, but it is laborintensive, requires immediate processing, and relies on a subjective interpretation of the slides (1, 7). Qualitative PCR detection of HCMV DNA in leukocytes or plasma is considered the most sensitive method, but it lacks specificity for the diagnosis of HCMV disease (2,6,16). Quantification of HCMV DNA has been proposed to be more specifically associated with the disease (1,18).…”