IgM antibody detection of ppUL80A and ppUL32 by immunoblotting: an early parameter for recurrent cytomegalovirus infection in renal transplant recipients.

The value of IgM detection for the early diagnosis of an active cytomegalovirus (CMV) infection in renal transplant recipients was evaluated prospectively. Sequential serum samples obtained from 22 allograft recipients with active CMV infection were tested for the presence of CMV-specific immunoglobulin M antibodies (IgM) by an enzyme-linked immunosorbent assay (ELISA) and a microparticle enzyme immunoassay (MEIA) and were compared with the Western-immunoblotting technique (IB). The time course of CMV IgM antibody detection was evaluated in relation to the shell vial assay (SVA), CMV disease, and immunosuppressive regimen. By IB, IgM antibodies against the capsid protein ppUL80a and the basic matrix phosphoprotein ppUL32 were detected in all 22 recipients with active CMV infection. Using the MEIA and the ELISA, the presence of CMV IgM antibodies was detected in 17 (77%) and ten (46%) of these 22 recipients, respectively. The SVA was the earliest parameter for detection of primary CMV infection in seven of nine (78%) recipients, in contrast to two of 13 (15%) patients with recurrent CMV infection (P < .05). The detection of IgM antibodies by IB was the earliest parameter for detection of recurrent CMV infection in seven out of 13 (54%) recipients in contrast to one out of nine (11%) patients with primary CMV infection (P < .05). During a primary CMV infection, the development of an abundant IgM antibody response was associated with recovery from CMV disease and the end of the viremic phase.

Risk factors for cytomegalovirus infection and disease in renal transplant recipients: HLA-DR7 and triple therapy.

In a prospective study, an analysis of risk factors for the development of cytomegalovirus (CMV) infection and disease was performed on 77 renal allograft recipients. Twenty-five out of the 77 recipients (32%) had a CMV infection. Twenty-two of the recipients received triple immunosuppressive therapy (cyclosporin A, prednisolone, and azathioprine) while the remaining 55 received standard therapy (cyclosporin A and prednisolone). In 23 recipients (30%) acute rejection was diagnosed and the first positive parameter of infection occurred 22 days after rejection therapy. Infection occurred in 10 out of 18 HLA-DR7-positive recipients (56%) and in 15 out of 59 HLA-DR7-negative recipients (25%; P < 0.02). In multiple regression analysis, HLA-DR7 was found to be a significant predictor of CMV infection (P < 0.005). CMV disease was diagnosed in only 9 out of 25 recipients with an acute infection. Six recipients (67%) with CMV disease received triple therapy for maintenance immunosuppression; this was significantly correlated to CMV disease (P < 0.05) as compared to three recipients (33%) with CMV disease maintained with standard therapy. Our data suggest that HLA-DR7-positive recipients are more susceptible to CMV infection and that CMV disease is associated with triple immunosuppressive therapy.

Cytomegalovirus IgM antibody detection: comparison of five assays.

Human Cytomegalovirus (CMV) infection can result in a dramatic disease in immunosuppressed patients. For effective treatment sensitive and specific procedures are required for prompt and early diagnosis of active CMV infection. In the present study five different serological assays were used for detection of CMV-specific immunoglobulin M antibodies (IgM) in 78 sera, obtained from patients with a clinical suspicion of CMV infection. In addition, sequential sera, obtained from renal transplant recipients who experienced a primary CMV infection, were analysed by the five IgM detection assays. The assays used were two enzyme-linked immunosorbent assays (ELISA): the Vironostika CMV ELISA from Organon Teknika (ELISA-1) and the Monosan CMV ELISA from Sanbio (ELISA-2), the CMV microparticle enzyme immunoassay (MEIA) from Abbott and two Western-(immuno)blotting techniques using purified CMV structural viral proteins of the AD169 strain (IB-AD169) and the major non structural DNA binding protein of 52 kiloDalton (IB-Rp52) as antigenic material. The results of the assays were compared to each other with respect to sensitivity, specificity and overall agreement, as well as incidence of false positive and false negative results. Although the MEIA gave the highest sensitivity, the ELISA-1 combined a high sensitivity with a high specificity and therefore appears the most suitable method for determination of active CMV infection.

Comparison of four techniques for detection of antibodies to cytomegalovirus.

Four serological methods were compared and evaluated for use in detecting cytomegalovirus antibody in blood and organ donors. Western blotting (immunoblotting), latex agglutination, enzyme-linked immunosorbent assay, and a recent available microparticle enzyme immunosorbent assay were used. The microparticle enzyme immunoassay appears to compare favorably with each of the other three assays tested for screening blood and organ donors for a previous cytomegalovirus infection.

Detection of latent human cytomegalovirus in organ tissue and the correlation with serological status.

The presence of human cytomegalovirus (HCMV) genome in spleen tissue was studied by using DNA hybridization techniques in seropositive and seronegative organ donors without clinical or laboratory confirmed HCMV infection. The serum samples of these patients were screened by latex agglutination test (LA) and enzyme linked immuno sorbent assay (ELISA) for the presence of HCMV antibodies, and confirmed by immunoblotting technique (IB). For the detection of HCMV sequences in spleen tissue dot blot DNA hybridization (DBH) using probes derived from immediate-early and late regions (ES and BH fragment respectively) of the HCMV genome were used. Samples positive in DBH were further tested by in situ DNA hybridization (ISH) using the ES probe. The number of spleen tissue specimens positive for HCMV nucleic acids indicated that HCMV may be present in human beings, even without serological evidence.