Cytomegalovirus DNA Levels Research Articles

Preemptive Therapy Prevents Cytomegalovirus End-Organ Disease in Treatment-Naïve Patients with Advanced HIV-1 Infection in the HAART Era

BackgroundThe efficacy of preemptive therapy against cytomegalovirus (CMV) infection remains unknown in treatment-naïve patients with advanced HIV-1 infection in the HAART era.MethodsThe subjects of this single-center observation study were126 treatment-naïve HIV-1 infected patients with positive CMV viremia between January 1, 2000 and December 31, 2006. Inclusion criteria were age more than 17 years, CD4 count less than 100/μl, plasma CMV DNA positive, never having received antiretroviral therapy (ART) and no CMV end-organ disease (EOD) at first visit. The incidence of CMV-EOD was compared in patients with and without preemptive therapy against CMV-EOD. The effects of the CMV preemptive therapy were estimated in uni- and multivariate Cox hazards models.ResultsCMV-EOD was diagnosed in 30 of the 96 patients of the non-preemptive therapy group (31%, 230.3 per 1000 person-years), compared with 3 of the 30 patients of the preemptive therapy group (10%, 60.9 per 1000 person-years). Univariate (HR = 0.286; 95%CI, 0.087–0.939; p = 0.039) and multivariate (adjusted HR = 0.170; 95%CI, 0.049–0.602; p = 0.005) analyses confirmed that CMV-EOD is significantly prevented by CMV preemptive therapy. Multivariate analysis showed that plasma CMV DNA level correlated significantly with CMV-EOD (per log10/ml, adjusted HR = 1.941; 95%CI, 1.266–2.975; p = 0.002). Among the 30 patients on preemptive therapy, 7 (23.3%) developed grade 3–4 leukopenia. The mortality rate was not significantly different between the two groups (p = 0.193, Log-rank test).ConclusionsThe results indicate that preemptive therapy lowers the incidence of CMV-EOD by almost 25%. Preemptive therapy for treatment-naïve patients with CMV viremia is effective, although monitoring of potential treatment-related side effects is required.

Frequent detection of cytomegalovirus (CMV) DNA in the lower respiratory tract in CMV‐seropositive pediatric patients with underlying chronic bronchopulmonary diseases lacking canonical immunosuppression

Cytomegalovirus (CMV) may be a relevant cause of morbidity in patients displaying various inflammatory diseases. In this study, it was investigated whether CMV DNA is detected in the lower respiratory tract and the systemic compartment in pediatric patients with chronic or recurrent bronchopulmonary diseases. A total of 42 lower respiratory tract specimens and 11 paired plasma samples from 42 patients were analyzed for the presence of CMV DNA by real‐time PCR. The respiratory specimens were also screened for the presence of respiratory viruses and human herpesvirus 6 (HHV‐6) and 7 (HHV‐7) by PCR methods. Quantitative bacterial and fungal cultures were performed. IL‐6 levels in the respiratory specimens were quantified using ELISA. CMV DNA was detected either in the lower respiratory airways, in plasma, or both in 54.5% of CMV‐seropositive patients. The levels of IL‐6 were significantly higher in these patients than in those with no detectable levels of CMV DNA. HHV‐6 and HHV‐7 DNA were detected in three and one patients, respectively. Respiratory viruses were detected in 13 of the 42 patients. Significant growth of one or more bacterial species was observed in 17 patients. No significant association was found between the presence of CMV DNA and the detection of other microorganisms. The data indicated that the presence of CMV DNA in the lower respiratory tract is a frequent finding in children with chronic or recurrent bronchopulmonary diseases. Further, prospective observational studies are needed to assess the impact of this phenomenon, if any, on the clinical course of these patients. J. Med. Virol. 85:888–892, 2013. © 2013 Wiley Periodicals, Inc.

Impact of seminal cytomegalovirus replication on HIV‐1 dynamics between blood and semen

The genital tract of individuals infected with human immunodeficiency virus type 1 (HIV-1) is an anatomic compartment that supports local HIV-1 and cytomegalovirus (CMV) replication. This study investigated the association of seminal CMV replication with changes in HIV-1 clonal expansion, evolution and phylogenetic compartmentalization between blood and semen. Fourteen paired blood and semen samples were analyzed from four untreated subjects. Clonal sequences (n = 607) were generated from extracted HIV-1 RNA (env C2-V3 region), and HIV-1 and CMV levels were measured in the seminal plasma by real-time PCR. Sequence alignments were evaluated for: (i) viral compartmentalization between semen and blood samples using Slatkin-Maddison and F(ST) methods, (ii) different nucleotide substitution rates in semen and blood, and (iii) association between proportions of clonal HIV-1 sequences in each compartment and seminal CMV levels. Half of the semen samples had detectable CMV DNA, with at least one CMV positive sample for each patient. Seminal CMV DNA levels correlated positively with seminal HIV-1 RNA levels (Spearman P = 0.05). A trend towards an association between compartmentalization of HIV-1 sequences sampled from blood and semen and presence of seminal CMV was observed (Cochran Q test P = 0.12). Evolutionary rates between semen and blood HIV-1 populations did not differ significantly, and there was no significant association between seminal CMV DNA levels and the frequency of non-unique clonal HIV-1 sequences in the semen. In conclusion, the effects of CMV replication on HIV-1 viral and immunologic dynamics within the male genital tract are not significant enough to perturb evolution or disrupt compartmentalization in the genital tract.

Immunoglobulin A-dominant postinfectious glomerulonephritis in a patient with acute HIV syndrome

Immunoglobulin A-dominant postinfectious glomerulonephritis in a patient with acute HIV syndrome

Comparison of real-time PCR quantitative analysis of the cytomegalovirus DNA level using LightCycler 2.0 and LightCycler 480 instruments

Cytomegalovirus infection is a frequent complication after transplantation. It is commonly believed that the level of CMV viraemia, usually measured with real-time PCR, is directly correlated with the risk of developing a serious cytomegaloviral disease. ObjectivesThe aim was the comparison of results obtained with a commercial test for the quantitative diagnostics of CMV infections, using LightCycler 2.0 and LightCycler 480 real-time PCR systems. Study designsStudy comprised 97 samples of nucleic acids isolated from serum, in which the CMV DNA was detected during routine tests. Measurement in both analysers was performed simultaneously for paired serum samples. Comparison and of the results obtained with two types of thermocyclers included regression analysis, Spearman correlation, Friedman test and Bland–Altman analysis (p<0.05). ResultsCMV DNA was detected in all samples with use of both thermocyclers. Correlation coefficient of results obtained with both analysers was 0.94. According to Friedman and Bland–Altman tests’ results, there were significant differences in measurements between instruments (p<0.001 and p=0.017, respectively). Difference of median values was 0.137log10CMV viral load/ml, and difference of mean values was 0.215log10CMV viral load/ml. Clear threshold of fluorescence crossing point of 1.4–1.8 cycles in advantage of LightCycler 480 was an apparent difference between both analysers. ConclusionsThere was good overall correlation between both analysers, as detected shift in both median and mean values was close to intra-assay variability declared by its producer. Observed differences were probably due to different fluorescence excitation and detection systems used in both instruments.

Can measurement of maternal anti‐cytomegalovirus immunoglobulin‐M antibody levels be used to screen for cytomegalovirus infection in embryos and fetuses?

Our study aims to estimate whether measurement of maternal anti-cytomegalovirus immunoglobulin-M antibody (CMV-IgM) levels are useful as a screening method for achieving early detection of congenital CMV infection. Levels of maternal CMV-IgM were measured by enzyme immunoassay in all (n =2865) pregnant women who visited our hospital in the first trimester during the period from January 2005 to December 2009. Among them, 21 individuals (0.73%) had a CMV-IgM titer of ≥0.08 and were judged to be CMV-IgM-positive. Informed consent was obtained from all 21 individuals to perform the confirmation test that quantifies the levels of cytomegalovirus DNA (CMV-DNA) in amniotic fluid by real-time polymerase chain reaction. However, only one (0.03%) of the 21 individuals was CMV-DNA-positive (CMV-DNA concentration, 1.0 × 10(4) copies/ml). In order to detect congenital CMV infection in early pregnancy, it is considered appropriate to use ultrasound for close examination of embryo or fetal symptoms indicative of CMV instead of performing serological screening based on CMV-IgM.

Risk Factors for Cytomegalovirus Retinitis in Patients with Cytomegalovirus Viremia after Hematopoietic Stem Cell Transplantation

To evaluate the risk factors for cytomegalovirus (CMV) retinitis in patients with CMV viremia after hematopoietic stem cell transplantation (HSCT). Retrospective cohort study. We included all patients with CMV viremia detected by polymerase chain reaction after HSCT between April 2009 and August 2011. Risk factors for CMV retinitis were evaluated in the cohort of 270 patients with CMV viremia, who survived ≥ 12 weeks after HSCT and were screened for CMV retinitis. Retrospective review of clinical records and laboratory results. Survival analysis of patients in the cohort and frequency of CMV retinitis in relation to various factors. Variables analyzed were demographics, human leukocyte antigen (HLA) matched versus mismatched, related versus unrelated donor, preconditioning regimens, delayed engraftment of lymphocyte, presence of acute or chronic graft-versus-host disease, highest CMV DNA level in blood (copies/ml), cumulative period of CMV viremia (weeks), and CMV infection verified by culture or immunohistology in bronchoalveolar lavage or visceral biopsy specimens. Of the 708 patients who underwent HSCT during the study period, 363 (51%) developed CMV viremia after HSCT. Of the 363 patients with CMV viremia, 270 underwent retinal examination for CMV retinitis. We detected CMV retinitis in 15 of 270 patients with CMV viremia. In the univariate analysis, HLA-mismatched HSCT, HSCT from an unrelated donor, engraftment day, peak CMV DNA level, and duration of viremia were associated with the development of CMV retinitis. In the adjusted multivariate analysis, only peak CMV DNA blood levels predicted the development of CMV retinitis (hazard ratio, 25.0; 95% confidence interval, 3.0-210.8). An additional validity analysis by receiver operating characteristic area under curve suggested that a cutoff of 7.64 × 10(4) copies/mL best predicted the development of CMV retinitis by CMV DNA levels in blood. The development of CMV retinitis should be carefully monitored in patients with a significant viral load, which is represented by a peak CMV DNA level >7.64 × 10(4) copies/ml and a long duration of CMV viremia, especially when patients received HSCT from an unrelated or HLA-mismatched donor and showed delayed lymphocyte engraftment.

Quantification of Epstein-Barr Virus DNA Is Helpful for Evaluation of Chronic Active Epstein-Barr Virus Infection

Chronic active Epstein-Barr virus infection (CAEBV) presents with chronic or recurrent infectious mononucleosis-like symptoms, such as low-grade fever, liver dysfunction, lymphadenopathy, and hepatosplenomegaly. Immunological methods are useful for the diagnosis of viral infections. However, CAEBV patients do not necessarily have high titers of Epstein-Barr virus (EBV)-specific antibodies. Hosts that are immunocompromised after hematopoietic stem cell transplantations sometimes suffer from systemic EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) and EBV-positive lymphoma. Patients with EBV-associated diseases are often diagnosed by analyses of bone marrow. Cytomegalovirus (CMV) can cause serious pneumonia or retinitis in immunocompromised hosts. In order to noninvasively understand the clinical status of patients with EBV-associated diseases, we conducted real-time polymerase chain reaction (PCR) methods in their peripheral blood in order to quantify EBV and CMV DNA levels, which reflect viral activity. Here, we describe a 30-year-old Japanese female patient with CAEBV. The patient had repeated fever, fatigue, and liver dysfunction. The histopathological results of liver biopsies were positive for EBV-encoded RNA-1. Acute hepatitis was associated with the EBV infection. The whole-blood EBV DNA levels were high and above 1.0 × 10⁷ copies/mL. After immunosuppressive and antiviral therapies, EBV DNA levels lowered. However, she had to receive bone marrow transplantation because of her EBV-HLH. As the number of lymphocytes increased in the post-transplantation period, EBV DNA levels gradually increased again. The simultaneous detection of CMV DNA was more sensitive than the CMV antigenemia test that is often used to diagnose CMV infections. Unfortunately, the patient died due to a fungal infection. Observing EBV DNA levels closely with real-time quantitative PCR methods is helpful for evaluating the changes in the clinical course.

Cytomegalovirus viraemia in the modern antiretroviral era*

Cytomegalovirus viraemia in the modern antiretroviral era*

Cytomegalovirus Viremia, Pneumonitis, and Tocilizumab Therapy

Cytomegalovirus Viremia, Pneumonitis, and Tocilizumab Therapy

High incidence of cytomegalovirus, human herpesvirus-6, and Epstein-Barr virus reactivation in patients receiving cytotoxic chemotherapy for Adult T cell leukemia

The etiology of cytomegalovirus (CMV), human herpesvirus-6 (HHV-6), and Epstein-Barr virus (EBV) reactivation and the potential for complications following cytotoxic chemotherapy in the absence of allogeneic transplantation are not clearly understood. Patients with adult T cell leukemia (ATL) are susceptible to opportunistic infections. In this study, the incidence, kinetics and clinical significance of reactivation of CMV, HHV-6, and EBV in ATL patients were investigated. Viral DNA in a total of 468 plasma samples from 34 patients was quantified using real-time PCR. The probability of CMV, HHV-6, and EBV reactivation by 100 days after the start of chemotherapy was 50.6%, 52.3%, and 21.6%, respectively. Although most CMV reactivations were self-limited, plasma CMV DNA tended to persist or increase if the CMV DNA levels in plasma reached ≥ 10(4) copies/ml. CMV reactivation was negatively associated with survival, but the P-value for this association was near the borderline of statistical significance (P=0.052). One patient developed fatal interstitial pneumonia concomitant with peak CMV DNA accumulation (1.6 × 10(6) copies/ml plasma). Most HHV-6 and EBV reactivations were self-limited, and no disease resulting from HHV-6 or EBV was confirmed. HHV-6 and EBV reactivation were not associated with reduced survival (P=0.35 and 0.11, respectively). These findings demonstrated that subclinical reactivation of CMV, HHV-6, and EBV were common in ATL patients receiving chemotherapy. There were differences in the viral reactivation patterns among the three viruses. A CMV load ≥ 10(4) copies/ml plasma was indicative of subsequent exacerbation of CMV reactivation and developing serious clinical course.

Cytomegalovirus infection and immunosuppressant treatment in allogeneic hematopoietic stem cell transplantation recipients

To explore the correlation between peripheral blood cytomegalovirus (CMV) DNA level and cyclosporine A (CsA) plasma concentration among allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients who received immunosuppressant treatment, and to evaluate the potential clinical value. A total of 32 allo-HSCT patients were enrolled and their data were analyzed retrospectively. Ganciclovir was used to prevent CMV infection before the transplantation. Routine fluorescence PCR was admitted to test the blood CMV DNA level. The patients were divided into 2 groups: a CMV DNA positive group and a CMV DNA negative group. Enzyme multiplied immunoassay technique was adopted regularly to monitor the blood CsA concentration. The correlation between CMV DNA level and CsA concentration was analyzed. The CMV infection rate in patients who received allo-HSCT was 53.13%. The blood CsA concentration in the CMV DNA positive group was significantly higher than that in the CMV DNA negative group (P<0.05). Through the ROC curve, the area under the curve on Day 1, 7, and 14 had statistical significance compared with 0.5, and the corresponding blood CsA concentration was 203.15, 215.55, and 302.65 ng/mL, respectively. Immunosuppressive drug concentration can affect the dynamic changes of CMV DNA. High blood CsA concentration may be one of the reasons for CMV infection. Monitoring the blood CsA concentration may provide guidance for clinical treatment.

Opportunistic virus DNA levels after pediatric stem cell transplantation: serostatus matching, anti‐thymocyte globulin, and total body irradiation are additive risk factors

Viral opportunistic infections remain a threat to survival after stem cell transplantation (SCT). We retrospectively investigated infections caused by cytomegalovirus (CMV), Epstein-Barr virus (EBV), human herpesvirus type 6 (HHV6), or adenovirus (AdV) during the first 6-12 months after pediatric SCT. Serum samples from 47 consecutive patients were analyzed by quantitative real-time polymerase chain reaction assay. DNAemia at any time point occurred for CMV in 47%, for EBV in 45%, for HHV6 in 28%, and for AdV in 28%. Three patients (6.3%) died of CMV-, EBV-, or AdV-related complications 4, 9, and 24 weeks after SCT, respectively, representing 21% of total mortality. These 3 cases were clearly distinguishable by DNAemia increasing to high levels. Serum positivity for CMV immunoglobulin G in either recipient or donor at the time of SCT, total body irradiation, and anti-thymocyte globulin conditioning were independent risk factors for high CMV or EBV DNA levels. We conclude that DNAemia levels help to distinguish significant viral infections, and that surveillance and prophylactic measures should be focused on patients with risk factors in whom viral complications rapidly can become fatal.

Virological and immunological features of active cytomegalovirus infection in nonimmunosuppressed patients in a surgical and trauma intensive care unit

Cytomegalovirus (CMV) reactivation occurs frequently in critically ill patients. The natural course of CMV infection and the interaction between CMV and the adaptive immune system in this setting remain poorly defined. Fifty-three CMV-seropositive patients in a surgical and trauma intensive care unit were included in this study. The CMV DNA load in tracheal aspirates (TA) and plasma (PL) was monitored by qPCR. CMV-specific T-cell immunity was assessed by intracellular cytokine staining. Plasma TNF-alpha levels were determined by ELISA. CMV reactivation occurred in 39.7% of patients (23% had CMV DNA detected only in TA). The analysis of TA allowed an earlier diagnosis in 28% of patients. Clearance of CMV DNAemia preceded that of CMV DNA in TA in some episodes. Peak CMV DNA levels were significantly higher in TA than in PL (P = 0.02). CMV reactivation developed in the presence of CMV-specific T cells. Termination of CMV reactivation was associated with an expansion of functional CMV-specific T cells. Plasma levels of TNF-alpha did not allow for the prediction of the occurrence of CMV reactivation. CMV-specific T-cell immunity is preserved in most critically ill patients experiencing CMV reactivation. Analysis of respiratory specimens is imperative for an optimal monitoring of CMV reactivation in this setting.

Surveillance of active human cytomegalovirus infection in hematopoietic stem cell transplantation (HLA sibling identical donor): search for optimal cutoff value by real-time PCR

BackgroundHuman cytomegalovirus (CMV) infection still causes significant morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Therefore, it is extremely important to diagnosis and monitor active CMV infection in HSCT patients, defining the CMV DNA levels of virus replication that warrant intervention with antiviral agents in order to accurately prevent CMV disease and further related complications.MethodsDuring the first 150 days after allogeneic HSTC, thirty patients were monitored weekly for active CMV infection by pp65 antigenemia, nested-PCR and real-time PCR assays. Receiver operating characteristic (ROC) plot analysis was performed to determine a threshold value of the CMV DNA load by real-time PCR.ResultsUsing ROC curves, the optimal cutoff value by real-time PCR was 418.4 copies/104 PBL (sensitivity, 71.4%; specificity, 89.7%). Twenty seven (90%) of the 30 analyzed patients had active CMV infection and two (6.7%) developed CMV disease. Eleven (40.7%) of these 27 patients had acute GVHD, 18 (66.7%) had opportunistic infection, 5 (18.5%) had chronic rejection and 11 (40.7%) died - one died of CMV disease associated with GVHD and bacterial infection.ConclusionsThe low incidence of CMV disease in HSCT recipients in our study attests to the efficacy of CMV surveillance based on clinical routine assay. The quantification of CMV DNA load using real-time PCR appears to be applicable to the clinical practice and an optimal cutoff value for guiding timely preemptive therapy should be clinically validated in future studies.

Detection and Comparison of Cytomegalovirus DNA Levels in Amniotic Fluid and Fetal Ascites in a Second-Trimester Fetus With Massive Ascites, Hyperechogenic Bowel, Ventriculomegaly and Intrauterine Growth Restriction

To present a prenatal diagnosis of congenital cytomegalovirus (CMV) infection in a pregnancy with fetal ascites. A 33-year-old, gravida 6, para 2, woman was referred to a hospital at 20 weeks of gestation for management of fetal ascites. The woman had not experienced recent rubella or herpes simplex infections. The maternal blood group was O and Rh(D)-positive. The maternal serum thalassemia and syphilis screen results were negative. Fetal ascites was first noted at 17 weeks of gestation. At 18 weeks, she underwent amniocentesis revealing a 46,XX karyotype. At 20 weeks of gestation, maternal serum CMV IgG and CMV IgM were positive. At 21 gestational weeks, prenatal ultrasound showed fetal ascites, hyperechogenic bowel, ventriculomegaly, and intrauterine growth restriction. Repeated amniocentesis showed CMV DNA levels of 9.72 x 10(5) copies/mL and 6.03 x 10(5) copies/mL in amniocytes and amniotic fluid supernatant, respectively. Paracentesis showed CMV DNA levels of 1.64 x 10(3) copies/mL and 114 copies/mL in ascitic cells and ascitic supernatant, respectively. The pregnancy was terminated. Postnatally, CMV DNA was detected in the umbilical cord, amnion, placenta, cord blood, lungs, liver and brain by quantitative real-time polymerase chain reaction. A prenatal diagnosis of fetal ascites in association with ventriculomegaly, hyperechogenic bowel and intrauterine growth restriction should alert physicians to congenital CMV infection in addition to aneuploidy. The present case provides evidence that CMV DNA levels are higher in amniotic fluid (amniocytes and amniotic fluid supernatant) than in ascites (ascitic cells and ascitic supernatant) in cases of congenital CMV infection.

The natural course of primary cytomegalovirus infection in blood donors

As cytomegalovirus (CMV) DNA is frequently detectable in the plasma of recently infected sero-positive blood donors, information concerning primary CMV infection is important for the identification of possibly infectious donors. Monitoring of 17 982 donors for CMV antibodies and DNA in plasma identified 14 subjects with ongoing primary CMV infection. Thirteen donors were interrogated for possible sources of infection and CMV-related symptoms, and monitored for CMV antigens, CMV DNA in plasma, leucocytes and urine, course of IgG and IgM antibodies as well as markers of systemic infection and parameters of organ function. CMV antigens and DNA were detectable in peripheral blood for up to 54 and 269 days respectively. Clearance of CMV DNA from blood correlated with clearance of IgM antibodies, development of IgG antibodies against the membrane glycoprotein gB and development of high avidity IgG antibodies. Eighty-five percent of subjects with primary CMV infection, but even 69% of matched controls reported possibly CMV-related symptoms. Sixty-two and 23%, respectively, had contact with possible sources of infection. One donor developed a febrile illness accompanied by increased levels of CMV DNA in peripheral blood 2 to 3 weeks after seroconversion. In other donors, neither markers of systemic infection nor parameters of organ function correlated with the course of CMV DNA and antigens. Potentially infectious donors can be identified by measuring CMV DNA, IgM antibodies or avidity of IgG antibodies. Alternatively, blood products donated during the first year after seroconversion should not be used for immunocompromised patients.

Primary cytomegalovirus infectious colitis complicating Crohn's disease successfully treated with oral valganciclovir

Most cases of cytomegalovirus (CMV) colitis that develop in patients with inflammatory bowel disease (IBD) are caused by reactivation of a latent virus. Primary CMV infections are rare in adult patients. Treatment with immunosuppressive agents increases the infection risk in patients with IBD. We present a 26 year old lady with primary CMV colitis, superimposed on underlying Crohn's colitis. The diagnosis was confirmed by a viral-like prodrome, a positive CMV IgM titer, presence of low avidity IgG antibodies to CMV, high CMV DNA titers in the plasma, and immunohistological detection of CMV positive cells in her colonic mucosa. The patient responded to initial treatment with intravenous ganciclovir with a fall in plasma levels of CMV DNA, treatment was completed with oral valganciclovir until plasma CMV DNA levels became undetectable.

Quantification of cytomegalovirus DNA levels in intestinal biopsies as a diagnostic tool for CMV intestinal disease

Background CMV intestinal disease (CMV-ID) is a serious complication in immunocompromised patients and mainly diagnosed by clinical, endoscopic and histopathologic findings, whereas qualitative CMV-PCR in tissue samples is not recommended for diagnosis due to its low positive predictive value (PPV). Objectives To study the interpretation and diagnostic use of CMV-quantification by PCR in intestinal tissue biopsies to recognize CMV-ID. To develop cut-off intestinal CMV-loads attributing illness to CMV. Study design CMV-genome copies in 163 biopsies from the lower intestinal tract of immunocompromised patients were determined by quantitative real-time PCR, normalized to the cell number, and retrospectively compared to histopathological analysis, clinical findings and occurrence of CMV-antigenemia. Two cut-off intestinal CMV-loads, cut-off histo and cut-off clin , were defined using histopathological or clinical criteria as gold standard, respectively. Results CMV was detected in 32.5% of biopsies with a more than six log range of CMV-concentrations (1 × 10 −4–1.4 × 10 2 copies/cell). Notably, biopsies with histopathologically or clinically confirmed CMV-ID had a significantly higher CMV-load ( p < 0.001). Cut-off histo and cut-off clin were defined at the intestinal CMV-load of 0.14 and 0.01 copies/cell, respectively, and improved the PPV. However, cut-off histo showed a decreased sensitivity for clinically defined CMV-ID cases. Interestingly, many patients with CMV-ID showed no concomitant CMV-antigenemia, suggesting a localized intestinal CMV-replication. Conclusions Quantification of CMV in intestinal biopsies is a useful diagnostic tool allowing the definition of cut-off values that can predict CMV-ID more accurate than qualitative PCR results. Further prospective studies have to clarify wether these cut-offs can improve diagnostics and treatment of CMV-ID in day-to-day clinical practice.

CMV DNA levels and CMV gB subtypes in ART-naive HAART-treated patients: a 2-year follow-up study in The Netherlands

In the pre-HAART period, HIV-1 patients were greatly at risk for cytomegalovirus (CMV) disease. In HAART-treated patients, the incidence of CMV disease has decreased dramatically and the timing and presentation of CMV infection may be different. Also the relevance of different CMV genotypes is part of debate. A total of 132 antiretroviral naive patients starting HAART were selected for a 2-year follow-up study in the Netherlands. In 105 (80%) patients, CMV DNA were less than 100 copies/ml in all plasma samples during follow-up. In 27 (20%) patients, a detectable CMV load was found during follow-up. In seven patients, the initial decrease in HIV-1 loads during HAART was accompanied by an increase in CMV loads. Of 1348 plasma samples, only 50 (3.7%) samples were positive with a CMV load more of than 100 copies/ml plasma. CMV loads more than 1000 copies/ml were found only in samples with CD4 levels less than 250 x 10 cells/l and with detectable HIV-1 loads. CMV glycoprotein B (gB) typing was possible in 19 patients. Among these patients, including four patients with triple CMV infection and seven patients with double infection, the most prevalent genotype was gB3 (16x) followed by gB2 (9x), gB1 (5x) and gB4 (4x). CMV disease during HAART is very unlikely as soon as the HIV-1 viral load becomes undetectable (<50 copies/ml) and/or CD4 cell levels are restored to more than 250 x 10 cells/l. Within Dutch HAART treated patients, infection with CMV gB3 is most prevalent, but also double or triple infection with other CMV gB strains are common.