Early diagnosis of cytomegalovirus hepatitis in liver transplant recipients: Role of immunostaining, dna hybridization and culture of hepatic tissue

Abstract

Immunostaining techniques that use a monoclonal antibody against an early cytomegalovirus antigen or a polyclonal antibody, in situ DNA hybridization and inoculation of cell cultures for the detection of cytomegalovirus from liver biopsy specimens were studied in 20 liver transplant patients with cytomegalovirus hepatitis, as defined by histological criteria. A total of 108 liver biopsy specimens from 20 patients with a diagnosis of cytomegalovirus hepatitis (obtained per protocol at 7, 21, 90, and 180 days or whenever liver dysfunction occurred), which had previously been examined histologically and in cell culture, were again studied by recutting the liver tissue for histological examination, DNA hybridization and immunostaining with monoclonal or polyclonal antibodies to cytomegalovirus. In 5 of 20 patients, the diagnosis of cytomegalovirus hepatitis could have been made earlier (mean = 9.6 days) by immunostaining with a monoclonal antibody. Of 47 biopsy specimens with cytomegalovirus inclusion bodies, the sensitivity and specificity of the diagnostic procedures were immunostaining with monoclonal antibody (84% and 90%) and polyclonal antibody (72% and 97%), in situ DNA hybridization (72% and 100%) and cell culture detection (52% and 95%), respectively. Immunostaining with a monoclonal antibody against an early CMV antigen frequently detected cytomegalovirus infection in the liver allograft earlier than identification of typical histological inclusion bodies. DNA in situ hybridization was less sensitive than other techniques but highly specific; cytomegalovirus cell culture lacked sensitivity compared with the other procedures.

Clinical Utility of Viral Load in Management of Cytomegalovirus Infection after Solid Organ Transplantation

“…Histopathology remains the reference standard for diagnosis of tissue-invasive CMV disease (2,3,54,70). CMV infection is indicated in a tissue biopsy specimen by cellular and nuclear enlargement (cytomegalic cells) and the presence of amphophilic to basophilic cytoplasmic inclusions (aggregates of CMV nucleoproteins that are produced during viral replication) (2,3,54,71). The severity of CMV infection can be assessed based on the degree of histological involvement.…”

“…While these histopathologic findings are highly characteristic of CMV infection (72), atypical features may be present and may overlap in appearance both with reactive changes and with inclusions of other intracellular viruses. Hence, the diagnosis can be confirmed further by in situ hybridization (ISH) or immunohistochemical (IHC) testing (71). To facilitate histopathological identification of CMV-infected cells in tissue specimens, ISH uses CMV-specific cDNA probes that bind to viral DNA in the cellular material (71).…”

Management of cytomegalovirus infection after liver transplantation

“…Sensitivity and specificity may be enhanced by complementary use of immunostaining and in situ hybridization. 49, 50 The risk of an invasive procedure to obtain tissue samples also needs to be considered.…”

Drug Resistance due to Mutation in UL 97 and Certain Other Specific Genes in Cytomegalovirus Strains: An Overview

“…The positive results correlate well with active HCMV infection of the organ, e. g. hepatitis, but the sensitivity of the histopathological finding is relatively low (Colina et al, 1995;Paya et al, 1989). Immunostaining with specific polyclonal or monoclonal antibodies against HCMV antigens has increased the sensitivity of the method compared to conventional staining (Barkholt et al, 1994;Colina et al, 1995;Paya et al, 1990). …”

“…ISH methods, described for the identification of viral DNA in infected cells, have also been employed to improve the histological diagnosis of infection (Barkholt et Paya et al, 1990). HCMV specific probe, usually labeled by biotin, is used in these assays.…”