Comparison of immunoblotting with other serological methods and virus isolation for the early detection of primary cytomegalovirus infection in allograft recipients

Abstract

Sequential specimens from nine allograft recipients were examined by using a variety of methods to detect primary cytomegalovirus (CMV) infection as rapidly as possible posttransplantation. Sera were examined for immunoglobulin G (IgG) and IgM antibodies by immunoblotting, enzyme immunoassay, and immunofluorescence and also by complement fixation, latex agglutination, and an immunofluorescence test for antibody to CMV early antigen. Urine and occasionally blood, tissue, and other specimens were centrifuged onto cell cultures to enhance CMV infectivity. Eight of the nine patients showed laboratory evidence of primary CMV infection, and CMV was isolated from seven of the eight: in no case was virus isolated before seroconversion had become evident. However, serological tests differed in their abilities to detect antibody response to CMV infection in different patients; while immunoblotting, latex agglutination, and enzyme immunoassay for IgG antibodies generally detected seroconversion before complement fixation, this was not invariably the case. At present, optimal laboratory detection of CMV infections in these patients can be achieved only by a combination of serological methods and virus isolation.

Temporal patterns of immunoblot-reactive antibodies to cytomegalovirus in transplant recipients

“…The patient groups in ourstudy produced antibodies to individual CMV antigens in a pattern similar to that displayed by immunocompetent patients with natural infection. Similar observations have already been made for renal [ 13,19,271 and bone marrow transplant recipients [35] but have not been reported for heart, liver, or small bowel transplant recipients. Proteins p150, p48, and p38 were the antigens most frequently detected by IgM and IgG in primarily and recurrently infected organ transplant recipients and in immunocompetent individuals, as has already been observed in immunocompetent individuals [9,171.…”

“…Using serological methods, detection of an active infection is based mainly on significant titer rises and, therefore, requires a consecutive serum, as opposed to the detection of specific IgM, which requires only one acute phase serum. Furthermore, seroconversion to CMV in some patients precedes or coincides with virus isolation [19,22].…”

Prospective study of cytomegalovirus antigenemia in allograft recipients

“…In many centers, patients are monitored for laboratory evidence of CMV infection following transplantation. However, the time-consuming nature and relative lack of sensitivity of classical methods for the detection of CMV are well documented, and multiple techniques must be used for maximum efficacy of monitoring, since no single standardized test can be relied upon for the earliest possible detection of CMV infections in all patients (5). Though polymerase chain reaction technology provides rapid and sensitive detection of the presence of viral genomes (3), its usefulness as a single monitoring test has not yet been demonstrated, and its application is, in any case, restricted by high cost and technical complexity.…”

“…Shell vial cultures of PMNLs and urine specimens were done in duplicate in human fetal lung fibroblasts as described by Smith and Espy (8), except that specimens were diluted 1:5 to avoid toxicity and staining was done at 3 days postinoculation, using the Syva Microtrak CMV Culture Identification Kit. In addition, duplicate tube cultures were inoculated and spin-amplified as described previously (5). Isolation of CMV was confirmed by IFA, using the Syva kit.…”

Physical and chemical methods for enhancing rapid detection of viruses and other agents.

“…Several review articles have reported the use of centrifugation for CMV diagnosis (106,107,265,266). The infectivity of human CMV was shown to be enhanced 8% by centrifugation of standard tube cultures at 2,000 x g for 1 h (cited in Miller et al [190]). The rapid detection of CMV by immune staining of cell cultures not centrifuged has been reported, and CMV can be detected directly in leukocytes and lavage specimens, etc.…”