Primary and recurrent cytomegalovirus infections have different effects on human herpesvirus-6 antibodies in immunosuppressed organ graft recipients: absence of virus cross-reactivity and evidence for virus interaction.

The relationship between serum antibodies to human herpesvirus-6 (HHV-6) and cytomegalovirus (CMV) infection was studied in immunosuppressed adult organ graft recipients all of whom had IgG to both HHV-6 and Epstein-Barr virus capsid antigen (EBVCA) before operation and who had received an organ or organs from HHV-6 seropositive donors. In primary CMV infection the titre of IgG to HHV-6 rose substantially (between 32- and 512-fold) in eight out of eight patients whereas IgG to EBVCA only rose 32-fold in two patients. Moreover, the HHV-6 responses coincided closely with the CMV seroconversion. Serum absorption studies gave no evidence for antibody cross-reaction between CMV and HHV-6 because the CMV antibody titre could be reduced specifically without affecting HHV-6 antibody titres and vice versa. In recurrent CMV infection, HHV-6 antibody levels rose (32-fold) in three out of eight patients but these changes did not coincide with the CMV antibody response. Similarly, in the complete absence of CMV infection, five out of eight patients showed antibody rises to HHV-6 (between four- and 16-fold). IgG titres to EBVCA were stable in both these groups of patients. It is concluded that there is serological evidence (rising titre greater than or equal to four-fold) for genuine HHV-6 reactivation or, alternatively, for reinfection in 16 out of the 24 patients. This phenomenon was most frequent in primary CMV infection where the largest HHV-6 antibody responses were seen probably because of an, as yet, undetermined interaction between the two viruses.     

Viral infections in renal transplant recipients.

Cytomegalovirus (CMV) infections are common in renal transplant recipients. We studied 23 recipients prospectively to determine whether infections by other herpes-group and non-herpes-group viruses were also present. Sera, obtained at the time of surgery and periodically thereafter, were tested for antibody to CMV, herpes simplex virus (HSV), Epstein-Barr virus (EBV), parainfluenza viruses types 1, 2, and 3, and the viruses of measles and rubella. We found no evidence of an unusual incidence of primary or secondary infection by the non-herpesviruses tested. Rises to CMV, HSV, and EBV antibody titers occurred in 43, 38, and 32% of patients, respectively. All serological rises to herpes-group viruses occurred in patients seropositive at the time of transplantation, with the exception of three patients who experienced primary CMV infections. We conclude that reactivation of all herpes-group viruses tested may occur in transplant recipients. Morbidity was associated only with primarly CMV infection.     

A mu-capture immunoassay for detection of human herpes virus-6 (HHV-6) IgM antibodies in human serum.

BACKGROUND: Human herpes virus-6 (HHV-6) was first isolated in 1986. It has been shown to cause exanthema subitum and has been associated with various other diseases. HHV-6 infection is widespread, and more than 90% of the population have antibodies against HHV-6 at the age of 2 years. Once acquired, the virus remains latent in the body. This makes it difficult to draw any conclusions about a causal relationship between the demonstration of HHV-6 and a specific disease. OBJECTIVES: This work was to develop a mu-capture HHV-6 IgM enzyme linked immuno sorbent assay (ELISA) for use in routine diagnosis and for wide scale patient population analysis. STUDY DESIGN: A mu-capture HHV-6 IgM ELISA was established. A total of 682 sera consisting of 585 sera from Danish blood donors and 97 sera from patients with autoimmune antibodies were analysed in the HHV-6 IGM ELISA. One hundred and ninety-two sera had earlier been analysed for total HHV-6 antibody content in a competitive ELISA, 94 sera were analysed for cytomegalovirus (CMV) IgM and 57 sera for Epstein Barr virus (EBV) antibodies, using different ELISA assays. The results for 12 primary infections with HHV-6 are also reported. RESULTS: A HHV-6 IgM optical density (OD)-ratio was calculated according to a constant positive control. An empirical cut off of 0.5 HHV-6 IgM OD-ratio was chosen (with regard to the 10 HHV-6 seroconverters), which resulted in a specificity of 97.5% of the HHV-6 IgM ELISA. Two of the three donor sera with HHV-6 IgM OD-ratios more than 1.05 had total HHV-6 antibody titers significantly above the group with IgM OD-ratios below 0.7 consisting with HHV-6 reactivation. There was no cross reactions to EBV or CMV IgM positive sera. CONCLUSION: The HHV-6 IgM ELISA seems valid to diagnose primary HHV-6 infection in particular in combination with the HHV-6 total antibody assay.     

A serological investigation of human herpesvirus 6 infections in liver transplant recipients and the detection of cross-reacting antibodies to cytomegalovirus.

Sera from 50 orthotopic liver transplant recipients were examined for antibodies to human herpesvirus 6 (HHV-6) and cytomegalovirus (CMV), and the findings correlated with the clinical condition of the patients. Both primary and secondary HHV-6 infections were detected serologically following liver transplantation. Interpretation of serological assays is complicated by CMV and HHV-6 antibody cross reactions which were common. Sera from 5 patients became HHV-6 antibody negative following absorption with CMV infected cells. Thirty patients were initially seronegative for HHV-6 antibodies, 12 remained so following transplantation, 5 developed cross reacting antibodies, and 13 seroconverted. The seroconversions occurred at 4 to 8 weeks post-transplant in the same time period as CMV antibody rises. HHV-6 IgM was detected in only 4 of the 13. Of the 7 patients who had serological evidence of active HHV-6 infections but no evidence of CMV infection, 4 (56%) had fever, 1 (14%) hepatitis, 1 (14%) lung dysfunction, and 3 (42%) neurological disorders. In the 12 patients who remained HHV-6 antibody negative, there were fewer fevers and neurological disorders.     

Cytomegalovirus infection in heart transplant recipients: Preliminary results of a controlled trial of intravenous gamma globulin

Cytomegalovirus (CMV) infection is a major problem during the first twelve weeks after cardiac transplantation. Reactivation of endogenous virus, blood transfusion, and the transplanted heart are sources for CMV infection. Not only can disseminated CMV infection contribute directly to mortality, but CMV infection predisposes to superinfection with bacterial and fungal pathogens. Studies in mice, the premature neonate, and bone marrow transplant recipients suggest that passive immunization with CMV antibody has a modifying effect on CMV infection. By prophylactic administration of intravenous immune globulin in a controlled randomized fashion to cardiac transplant recipients we will address the question of whether passively acquired antibody prevents or modifies infection with CMV or other viral, bacterial, and fungal infections. Thirteen patients undergoing cardiac transplantation at Stanford University were randomized to receive or not receive intravenous immune globulin (IGIV) at a dose of 20 ml/kg twenty-four hours post transplantation and then weekly thereafter for a period of 10 weeks. Six patients seronegative for CMV antibody were randomized separately from seven who were seropositive for CMV antibody. Seven patients randomly selected from these two groups received IGIV. In patients seronegative for CMV, administration of IGIV resulted in serum titers of 1:10,000–1:30,000 measured by radioimmunoassay. Antibody titers were maintained at levels of 1:10,000 to 1:30,000 throughout the 10 week period post transplantation. Antibody remained detectable up to 8 weeks after the last infusion. No significant increases in CMV antibody levels were detectable after IGIV administration in patients seropositive for CMV antibody prior to transplantation. The administration of IGIV in thirty-two large and repeated doses was safe and well tolerated in cardiac transplant recipients. Serum levels of CMV antibody comparable to those observed in normal CMV seropositive individuals were achieved in CMV seronegative transplant recipients and were maintained throughout the period of greatest risk for CMV infection.     

Effect of primary Epstein-Barr virus infection on human herpesvirus 6, cytomegalovirus, and measles virus immunoglobulin G titers.

Immunoglobulin G antibody titers to human herpesvirus 6 (HHV-6), measles virus, and cytomegalovirus (CMV) were examined in serum samples from 31 patients with Epstein-Barr virus (EBV)-induced infectious mononucleosis (IM). Sera were drawn sequentially from the same patients less than or equal to 7 days until 3 years after onset of IM. In seropositive patients, there was a significant decrease with time after IM of the immunoglobulin G titers to the three viruses in the majority of patients; HHV-6 IgG titers decreased in 80%, measles virus IgG titers decreased in 75%, and CMV IgG titers decreased in 67%. Four patients contracted CMV infection during the observation period after IM. In these, HHV-6 IgG titers increased, while EBV and measles virus IgG titers remained essentially stationary. Polyclonal B-cell stimulation during IM is suggested to augment antiviral titers in general, but the increases of HHV-6 IgG titers during EBV and CMV infections may also be due to selective stimulation of memory B cells by related antigens or to reactivation of HHV-6 during infection with these herpesviruses.     

Immunoglobulin M to cytomegalovirus in primary and reactivation infections in renal transplant recipients.

Two commercially available enzyme immunoassays and one assembled in house were used to measure immunoglobulin M (IgM) antibody to cytomegalovirus (CMV) in a total of 220 serum specimens from 104 renal transplant recipients. All assays included a step in which interfering IgG antibody was removed or complexed. Concordance of results between pairs of assays ranged from 84 to 96%. All sera from patients with recent seroconversion (primary CMV infection) had measurable anti-CMV IgM. Among those already seropositive to CMV when transplanted, 26 to 55% had IgM antibody posttransplant, depending on the assay. This was observed regardless of the CMV serologic status of the kidney donor, indicating that reactivation of endogenous CMV, as well as reinfection, can induce this antibody in transplant recipients. Four cadaver donors known to transmit CMV to eight recipients did not have measurable IgM antibody to CMV.     

Evaluation of anti-complement immunofluorescence test in cytomegalovirus infection.

The anti-complement immunofluorescence (ACIF) technique was evaluated for the diagnosis of human cytomegalovirus (CMV) infection in a group of sera derived from renal transplant recipients and donors by comparing it with the indirect immunofluorescence (FA) and complement fixation (CF) TESTS. The ACIF and FA tests yielded similar results. However, the ACIF test had a distinct advantage over the indirect FA test, since it eliminated the nonspecific cytoplasmic staining that may result in false positive readings in inexperienced hands. Both the indirect FA and ACIF tests were more sensitive than the CF test. In primary CMV infection, the FA and ACIF antibodies appeared earlier and had significantly higher titer than corresponding CF titers. This difference in titers was not seen in seropositive individuals who lacked overt infection. Our previously reported correlation between the seropositivity of the donor and CMV infection in seronegative recipients has been confirmed.     

Latex agglutination and enzyme-linked immunosorbent assays for cytomegalovirus serologic screening of transplant donors and recipients.

The effectiveness of three serologic assays (two enzyme-linked immunosorbent assays [ELISAs] and latex agglutination) for cytomegalovirus (CMV) serologic matching of donors and recipients was assessed over a 2-year period in a major organ transplant program. Sera with equivocal test results were investigated by repeat testing of serum samples and additional specimens from the individuals involved and monitoring of CMV infection in recipients. An in-house ELISA identified all CMV-infective donors as seropositive. Of 63 ELISA-positive donors, 5 were negative by latex agglutination; recipients from 3 of these donors developed primary CMV infection posttransplant. The in-house ELISA and a commercial ELISA (Abbott enzyme immunoassay; Abbott Laboratories, North Chicago, Ill.) had a 93% concordance of results; follow-up testing indicated that the Abbott assay was sensitive but had a false-positive rate of about 11%. One recipient with a false-positive result developed symptomatic primary CMV infection after receiving a seropositive organ. Thus, performance characteristics of currently used screening assays affect recipient outcome.     

Enzyme-linked immunosorbent assay for detection of antibody to human herpesvirus 6.

The results obtained with an enzyme-linked immunosorbent assay (ELISA) for detection of human herpesvirus 6 (HHV-6) immunoglobulin G using a single 1:100 dilution of serum correlated well with those found by an indirect fluorescence microscopic assay (IFA) (r = 0.71). Concordant results were found in all 7 paired serum samples obtained from patients with acute primary infections and in 37 of 41 (90.24%) single serum samples. Fourteen serum samples (25%) which yielded nonspecific results by IFA were evaluable by ELISA. In a serologic survey using the ELISA, a disproportionate number of 12-month-old infants had low difference-of-optical-density values, suggesting that maternal antibody might persist beyond a year of age. This finding and the rises in antibody to HHV-6 found in patients with primary cytomegalovirus infections might lead to overestimation of HHV-6 infection rates in young children in seroprevalence studies. Other herpesvirus infections produced lesser effects on anti-HHV-6.     

Cytomegalovirus infections in heart and heart and lung transplant recipients.

Of the first 166 heart and 15 heart and lung transplant recipients at Papworth Hospital, Cambridge, who survived for more than one month after transplantation, 162 were investigated for cytomegalovirus (CMV) infection by serological methods. Altogether, 73 (45%) developed CMV infection after transplantation: 30 (18.5%) had acquired primary infection and 43 (26.5%) reactivation or reinfection. Six patients died of primary infection, probably acquired from the donor organ. Recipients negative for CMV antibody who received an organ from an antibody positive donor had the most severe disease. Heart and lung transplant recipients experienced more severe primary CMV infection than those in whom the heart alone was transplanted. The most sensitive and rapid serological method was a mu-capture enzyme linked immunosorbent assay (ELISA) for detecting CMV specific IgM, the amount of which was often of prognostic value and influenced the management of patients.     

Activation of human herpesvirus-6 in children with acute measles.

Virological and serological studies were carried out prospectively to evaluate the possible activation of human herpesvirus-6 (HHV-6) in 50 infants and children with acute measles by isolation of HHV-6 from peripheral blood and by determining neutralizing antibodies to the virus. All but 5 patients (90%) were seropositive to HHV-6 in the acute stage of measles and 18 (40%) had a significant increase in HHV-6 antibody titers thereafter, whereas only 2 of 27 patients who were initially seropositive to Epstein-Barr virus (EBV) viral capsid antigen (VCA) had a significant rise in antibody titers to EBV VCA. Among 18 patients with a significant increase in HHV-6 titers, the virus was isolated from the peripheral blood mononuclear cells of three patients in the early convalescent stage of measles. These results indicate that activation of HHV-6 may occur frequently a few weeks after primary infection with the measles virus.     

HHV-6 and HHV-7 antigenemia related to CMV infection after liver transplantation

BACKGROUND: Betaherpesviruses human herpesvirus-6 and -7 (HHV-6, HHV-7), which are closely related to cytomegalovirus (CMV), have been reported in transplant patients. In this retrospective study, we investigated the occurrence of HHV-6 and HHV-7 antigenemia in relation to symptomatic CMV infection after liver transplantation. METHODS: Sample material from 64 adult liver transplant recipients was included in the study. The patients were monitored weekly for CMV, HHV-6, and HHV-7. CMV infections were diagnosed by pp65-antigenemia and viral cultures. Concomitantly HHV-6 and HHV-7 antigens were demonstrated in peripheral blood mononuclear cells by monoclonal antibodies against both variants A and B and immunoperoxidase staining. Altogether 540 post-transplant blood specimens were analyzed. RESULTS: Nineteen patients (30%) developed symptomatic CMV pp65 antigenemia during the first 3 months (mean 33 days, range 5-62 days) post-transplantation and were treated with intravenous ganciclovir. Concurrent HHV-6 antigenemia was detected in 16/19 (median 9 days, range 6-24 days) and HHV-7 antigenemia 15/19 patients (median 17 days, range 5-58 days) after transplantation. HHV-6 appeared before CMV in most cases (12/16), HHV-7 usually together with CMV. In those cases that HHV-6 preceded CMV antigenemia, it also was a possible cause of graft dysfunction. HHV-7 and CMV were so closely overlapping, that no symptoms could solely be linked with HHV-7. CONCLUSION: HHV-6 and HHV-7 antigenemia usually occurred together with symptomatic CMV infection after liver transplantation. HHV-6 preceded CMV, but HHV-7 appeared together with CMV. Further investigation of the clinical significance of HHV-6 and HHV-7 antigenemia in organ transplant patients is necessary.     

Longitudinal analysis of human herpesvirus-8 DNA and antibodies in an Italian allogeneic stem cell transplant recipient

Background

Changes of HHV-8 antibody reactivity and intermittent detection of HHV-8 DNA have been observed in subjects with Kaposis's Sarcoma and/or HIV infection. Little is known about the longitudinal dynamics of HHV-8 DNA and antibody response in allografted stem cell transplant (SCT) patients without Kaposis's Sarcoma.

Objectives

To report the natural history of a HHV-8 seropositive patient with chronic lymphocytic leukemia who developed an active HHV-8 infection after SCT.

Study design

HHV-8 antibodies were measured by IFA and ELISA assays. HHV-8 DNA was detected by real-time PCR quantitative assay in serum and peripheral blood leukocytes (PBL).

Results

Twenty-two out of 26 (85%) serum samples had detectable HHV-8 antibodies: 21/26 (80%) samples were positive by both IFA and ELISA assays, while 1 sample was ELISA positive-IFA negative. The remaining 4 samples (15%) were negative by both assays. Five out of 6 (83%) serum-PBL samples pairs had detectable HHV-8 DNA: a median of 934 genomes/ml (range 254–6316 genomes/ml) in the serum and a median of 10,000 genomes/10⁵ (range 1472–93,460 gen/10⁵) in PBL. An active HHV-8 infection occurred early within the first 30 days after the transplant, extended up to day +180 and occurred without evidence of HHV-8-related neoplastic or non neoplastic diseases.

Conclusions

This study provides evidence that a patient infected with HHV-8 before SCT can be either intermittently HHV-8 DNA-positive and/or seropositive for HHV-8 after SCT. A high and persistent HHV-8 replication may be ongoing even in the absence of overt HHV-8-associated diseases.     

Human cytomegalovirus (HCMV)-specific immunoglobulin E as a serologic marker for HCMV infection in immunocompromised patients

Summary An antibody capture assay using an enzyme-linked human cytomegalovirus (HCMV) antigen for the detection of specific immunoglobulin E (IgE) was established. IgG, M, and E responses to HCMV were studied in 497 sera obtained from 44 renal transplant recipients and 51 acquired immunodeficiency syndrome (AIDS) patients. The results were compared with those obtained from 58 HCMV-seropositive healthy individuals. HCMV-specific IgE was detected in 11 (91.7%) renal transplant recipients with primary HCMV infection. In contrast, antibodies of the IgG and IgM classes were detected in only 6 (50.0%) of these patients. Specific IgE was detected in 10 (90.9%) out of 11 renal allograft recipients suffering from secondary HCMV infection. Significant IgG titer rises and IgM were detected in 2 (18.2%) and 6 (54.6%) of these patients, respectively. IgG titer rises and IgM and IgE antibodies were seen in 5 (12.2%), 1 (2.4%) and 18 (43.9%) AIDS patients respectively. All healthy immunocompetent HCMV-seropositive individuals were tested IgE negative. The results obtained in our study indicate that IgE against HCMV is a more reliable serologic marker for primary and secondary HCMV infection than IgM in immunocompromised individuals, especially in organ transplant recipients, since it is not affected by the prophylactic application of HCMV hyperimmune globulin preparations.Abbreviations AIDS acquired immunodeficiency syndrome - BAL bronchoalveolar lavage - CDC Centers for Disease Control, Atlanta, USA - ELISA enzyme-linked immunosorbent assay - HCMV human cytomegalovirus - HIV human immunodeficiency virus - Ig immunoglobulin - PBS phosphate buffered saline - RTR renal transplant recipients     

IgG antibodies to human herpesvirus-6 in children and adults and in primary Epstein-Barr virus infections and cytomegalovirus infections [corrected

Antibody titers against human herpesvirus-6 (HHV-6) were determined in 80 healthy adults and 100 children and teenagers from Sweden to gain information on the role of the virus and its epidemiology. Based on a positive immunofluorescence titer of 1:10 and above, about 85% of the adults and children were seropositive with 60% seropositivity of children below age one year. Titers were generally higher in patients with simultaneous EBV or CMV infection, yet crossreactivity appeared essentially no problem. HHV-6 thus is ubiquitous like other herpesviruses. Primary infection seems to occur early in life, and reactivation or delayed primary infection may be associated with a variety of disorders.     

Analysis of the shedding of three β-herpesviruses DNA in Polish patients subjected to allogeneic hematopoietic stem cell transplantation: Six-year follow up

BackgroundInfections with human β-herpesviruses are common worldwide and are still frequent in patients after hematopoietic stem cell transplantation. Some data suggest that HHV-6 and HHV-7 could take part in CMV reactivation from latency and/or progression of CMV disease in immunosupressed patients.ObjectivesThe aims of this study were: (1) to summarise retrospectively the results of β-herpesviruses DNA detection in a large group of adult allogeneic haematopoietic stem cell transplant recipients; and (2) to find a potential correlation between viruses belonging to this subfamily.Study designAlloHSCT recipients (N = 142) were examined in the early post-transplant period (median = 89 days). The presence of CMV, HHV-6 and HHV-7 was confirmed through detection and quantification of viral DNA, isolated from 1679 sera samples.ResultsCMV DNA alone was detected in 23.9% of patients, while single HHV-6 and HHV-7 were detected in 14.8% and 9.9% of individuals, respectively. The reactivation of more than one virus was identified in 31% of analysed patients. In cases of concurrent infection, HHV-7 was detected at the same time as HHV-6, and both of them were usually reactivated before CMV. The kinetics of virus reactivation and measured viral load may suggest a potential role of HHV-6 and HHV-7 as co-factors in CMV reactivation.ConclusionsThe observed kinetics of virus reactivation may strongly suggest a potential role of HHV-6 and/or HHV-7 as co-factors of CMV reactivation. The co-infection with these β-herpesviruses could predispose patients after hematopoietic stem cell transplantation to a longer and more severe CMV infection.     

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. © 1996 Wiley-Liss, Inc.     

Analysis of antibody assay methods and classes of viral antibodies in serodiagnosis of cytomegalovirus infection.

Forty-nine serum pairs with antibody to cytomegalovirus (CMV) were evaluated for rises in antibody titer (greater than or equal to fourfold) by indirect hemagglutination (IHA) and complement fixation (CF), using a freeze-thaw antigen (FT) and a glycine extract antigen (GE). In this sample CF-FT detected more rises in antibody titer than did CF-GE. IHA detected the least number. The apparent reason for stationary antibody titers with CF-GE and IHA was the presence of high antibody titers in the first serum specimen. Separation of immunoglobulin classes of 20 serum pairs by sucrose gradient centrifugation indicated that these antibodies with IHA were of the immunoglobulin M (IgM) class and those with CF-GE were of the IgG class. By separation of immunoglobulin classes, rises in IgG CMV antibody titers were seen with IHA, rises not observed in the whole serum because of high IgM antibody titers in the first serum specimen. Absence of rises in antibody titers with CF-FT was due in part to too early sampling of the second serum specimen (less than 21 days) and in part to an apparent inability of some individuals to respond with antibody reactive with FT antigen. CF-GE and CF-FT antibodies of the IgM class were detected in some sera, usually in specimens collected more than 10 days after the onset of symptoms. Although reactive with CMV antigen, the specificity of these IgM antibodies in relation to rheumatoid factor requires clarification.     

Dynamic of immune response in primary cytomegalovirus infection and after reactivation of cytomegalovirus in patients with organ allotransplantation

A total of 44 serum specimens from 7 patients with kidneys or liver transplanted from donors who had antibodies (Ab) to human cytomegalovirus (CMV) were studied. In 4 recipients anti-CMV Ab were found before transplantation and in 3 others they were not detected. It was shown by EIA that IgM and IgG anti-CMV appeared in the sera of primarily infected patients after 1-2 weeks and their titers were 5-10 times lower than in patients with reactivated CMV infection. Immunoblotting of Ab to individual CMV proteins showed a narrower spectrum of Ab during the initial period of primary CMV infection in comparison with the same period of reactivation and delayed production of AB to conformation-dependent determinants. Hence, analysis of anti-CMV Ab during the first 4-6 weeks after organ transplantation by EIA and immunoblotting differentiates primary CMV infection from its reactivation by Ab titers and spectrum. These parameters vary in some patients.