Rapid detection of infectious cytomegalovirus in blood with the aid of monoclonal antibodies

Abstract

The recently developed early antigen immunofluorescence (IF) method for the detection of infectious cytomegalovirus (CMV) in clinical specimens has hardly been applied on blood samples. We compared the CMV early antigen detection technique with the conventional cell culture method in 415 different buffy coat samples from 85 different immunocompromised patients. Duplicate coverslips were stained with two different monoclonal antibodies 4-6 days after inoculation. The conventional cultures were examined for typical cytopathic effects (CPE) during 10 weeks. Forty samples from 19 patients were positive by the IF technique, most of them with both monoclonal antibodies. Only 22 of these samples were positive in the conventional cell culture assay, on average after 15.8 days. CMV viraemia was detected exclusively by the IF method in 18 samples, 7 of which were from five patients without any further evidence of an active CMV infection. CMV viraemia was detected exclusively by the CPE method in eight samples, on average after no less than 36.6 days. CMV viraemia was not found in blood samples from 10 patients with laboratory proven active CMV infections and 53 patients without any evidence of an active CMV infection. In our hands the early antigen method for the detection of infectious CMV in blood is nearly as specific (at least 98.1%) and clearly much faster and more sensitive than the conventional cell culture method. The early CMV antigen detection method is therefore a very useful tool for the rapid detection of infectious CMV in blood.

New Strategies for Prevention and Therapy of Cytomegalovirus Infection and Disease in Solid-Organ Transplant Recipients

“…CMVtriggered immune reactivity has been proposed, but sound evidence to support this is lacking (184,427). Once CMV is activated and even before clinical manifestations occur, infection of circulating leukocytes, particularly granulocytes, can be demonstrated (398,408,479). Infected leukocytes presumably serve as means of transporting this highly cell-associated virus to other body sites.…”

Quantitative Real-Time Polymerase Chain Reaction for Evaluating DNAemia due to Cytomegalovirus, Epstein-Barr Virus, and BK Virus in Solid-Organ Transplant Recipients

“…A very general approach is to introduce commercial standards of EBV DNA into the assay as described for analysis of CMV DNA [33]. At our institution, 95% of the EBV DNAemias were р copies/mL and 50% of the 4 5.5 ϫ 10 reported values were р EBV DNA/mL. 3 2.0 ϫ 10 Comprehensive studies are lacking for evaluation of the most appropriate compartment of blood (e.g., whole blood vs. plasma) to monitor EBV DNA in viremic patients.…”

“…BKV and EBV are not detected in cell cultures by routine diagnostic virology methods. Detection of CMV in blood specimens can be qualitatively recognized rapidly (16 h after inoculation) in shell vial cultures, but quantitation of the CMV load with this technology is cumbersome and impractical.The first laboratory method for quantitation of CMV load was the antigenemia test [4] jective interpretation of the test results [5][6][7]. Importantly, both shell vial cell cultures (qualitative) and antigenemia tests (quantitative) have been shown in comparative studies to be less sensitive than PCR.…”

Evidence for transfer of cellular and humoral immunity to cytomegalovirus from donor to recipient in allogeneic bone marrow transplantation

“…Specific binding was visualized by peroxidase staining (goat anti-mouse peroxidase-labelled IgG (TAGO, Burlingame, CA). 3-Amino-9-ethylcarbazole (AEC; Sigma Chemical Co., St Louis, MO) was used as substrate [11].…”