Presence Of Cytomegalovirus DNA Research Articles

Cytomegalovirus DNA load monitoring in stool specimens for anticipating the occurrence of intestinal acute graft-versus-host disease following allogeneic hematopoietic stem cell transplantation: Is it of any value?

Data have been published suggesting a bidirectional interaction between cytomegalovirus (CMV) infection and acute graft-versus-host disease (aGvHD) in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients. Here, we hypothesized that prospective CMV DNA monitoring in stool specimens may be useful for predicting subsequent occurrence of intestinal aGvHD (IaGvHD). This two-center study enrolled 121 consecutive adult patients undergoing any modality of allo-HSCT. A total of 1,009 stool specimens were collected (a median of 7 specimens/patient; range, 1-18). CMV DNA monitoring in stools and plasma was performed using real-time PCR assays. CMV DNA was detected in stools in 20 patients (cumulative incidence, 16.9%; 95% CI, 6.3%-31.8%). Median CMV DNA level in stool specimens was 1,258IU/0.1g (range, 210-4,087IU/0.1g). All these patients and their donors were CMV seropositive, and 16 of the 20 patients also had CMV DNAemia, while 4 patients had CMV DNA detected in stools without CMV DNAemia. No correlation was found between CMV DNA loads in plasma and stools (P=.40). Prior CMV DNAemia, aGvHD, or IaGvHD were not associated with presence of CMV DNA in feces. IaGvHD was present in 30 patients, in 5 of whom CMV DNA was detected in stools. Neither detection of CMV DNA in feces nor in plasma was associated with subsequent IaGvHD (OR, 0.67; 95% CI, 0.18-2.52; P=.55 and OR, 0.86; 95% CI, 0.38-1.96; P=.71, respectively). No patient in this cohort had CMV end-organ disease within the study period. Our study failed to provide evidence pointing to a reciprocal interaction between GI CMV infection and IaGvHD. CMV DNA monitoring in stools seems of no value to anticipate occurrence of IaGvHD.

Sex Differences in CMV Replication and HIV Persistence During Suppressive ART.

BackgroundThe association between subclinical cytomegalovirus (CMV) replication and HIV persistence has not been investigated in cis-gender women with HIV.MethodsFifty virologically suppressed female participants with HIV were prospectively enrolled and provided oral, vaginal, and urine samples and peripheral blood mononuclear cells at 1 cross-sectional time point. CMV DNA was quantified in each specimen by real-time polymerase chain reaction (PCR). Cellular HIV DNA and HIV RNA transcripts (unspliced and multiply spliced [ms] encoding tat-rev) were quantified by droplet digital (dd) PCR in peripheral blood cells. Forty-nine male individuals with HIV and CMV (historical data) were used as controls.ResultsLevels of cellular HIV DNA and unspliced HIV RNA were not different between sexes, but female participants had less detectable msHIV RNA and CMV DNA compared with males (both P < .01). Unlike previously described for males, the presence of CMV DNA was not associated with increased HIV DNA in females. Among female participants, premenopausal status was independently associated with lower HIV DNA compared with postmenopause, after adjusting for nadir CD4 count (P < .01).ConclusionsFemale participants with HIV had reduced cellular HIV RNA and less subclinical CMV DNA compared with males but overall similar HIV DNA levels in our study. Postmenopausal status was independently associated with higher HIV DNA levels among female participants.

Correlation the Cytomegalovirus (CMV) IgM Antibodies and Viral DNA Presence with Rheumatoid Arthritis (RA) in Iraqi Patients

In the Iraqi population, a high incidence (82.7%) of rheumatoid arthritis (RA) has been reported among the suspected patient. Many investigators studied the microbial infectious present in Iraqi patients with RA; however, to the best of our knowledge, there is no previous study detected the CMV DNA and antibodies to RA disease. Hence, the current study aimed to investigate the presence of CMV DNA and antibodies in Iraqi RA patients. A total of 58 blood samples were collected from patients with clinical signs of rheumatoid arthritis, along with 32 samples of apparently healthy individuals as a control group. These samples were tested for rheumatoid factor (RF), CMV IgM antibodies and viral DNA during the acute and the chronic periods of the autoimmune disease. The results showed that 46.6% (27/58) and 13.8% (8/58) of rheumatoid arthritis patients had positive reactions for IgM-CMV and CMV DNA, respectively, as compared to healthy individual. The highest rate of the viral occurrence was recorded in the aged and female RA patients (24 and 21 cases, respectively). Moreover, the most significant increases in RA appearance and CMV reactivation cases were observed in patients with a mean age higher than 40 years old. This finding pointed out the prevalence of viral infections mostly in RA patients aged more than 40 years old, accompanied by increased RA cases. This study concludes that patients with positive CMV must be tested for rheumatoid factor, especially when the viral DNA is present. Moreover, we recommend measuring TNF-α and IFN-γ as the proinflammatory cytokines that play an important role to protect against viral infection and prevent their reactivation in all RA patients.

Prevalence of cytomegalovirus DNAemia and genotypic distribution among childbearing mothers and neonates in Taiwan

BackgroundCongenital cytomegalovirus (CMV) infection is the leading cause of neurologic disabilities and sensorineural hearing loss in children. However, in Taiwan, there is limited information on the genotypic diversity and prevalence of perinatal CMV infection in both mothers and neonates. The aim of this study was to screen samples from both mothers and umbilical cord blood for CMV at the time of delivery and to determine the CMV genotypic distribution. MethodsBetween June 2012 and July 2015, residual maternal and umbilical cord blood samples were collected from consenting participants admitted to the Chang Bing Show Chwan Memorial Hospital in central Taiwan. The blood samples were screened for CMV DNA using real-time PCR assay, and the genotypic classification of the CMV UL55, UL144, and US28 genes was determined by sequencing and phylogenetic analysis. ResultsA total of 1282 mother-neonate paired samples were enrolled in the study, 95.3% of whom were Taiwanese. CMV DNA was detectable in 6.2% of the maternal blood samples, with a significantly higher rate noted in non-Taiwanese mothers (11.7%,p=0.027). For the 1,282 umbilical cord blood samples, CMV DNA was detectable in 5.3% of the samples. The presence of CMV DNA in maternal blood was positively associated with the presence of CMV DNA in umbilical cord blood (p=0.01). In addition, the UL55, UL144, and US28 genotypic distribution was similar between mothers and neonates. ConclusionThe prevalence of CMV DNAemia in childbearing mothers and neonates is similar and their genotypic distribution implies potential CMV infection during pregnancy.

The Detection of CMV in Saliva Can Mark a Systemic Infection with CMV in Renal Transplant Recipients.

Human cytomegalovirus (CMV) is often transmitted through saliva. The salivary gland is a site of CMV replication and saliva can be used to diagnose congenital CMV infections. CMV replication is monitored in whole blood or plasma in renal transplant recipients (RTR) and associates with clinical disease. However, these assays may not detect replication in the salivary gland and there is little data linking detection in saliva with systemic infection and clinical sequelae. RTR (n = 82) were recruited > 2 years after transplantation. An in-house quantitative PCR assay was used to detect CMV UL54 in saliva samples. CMV DNA was sought in plasma using a commercial assay. Vascular health was predicted using flow mediated dilatation (FMD) and plasma biomarkers. CMV-reactive antibodies were quantified by ELISA and circulating CMV-specific T-cells by an interferon-γ ELISpot assay. Vδ2− γδ T-cells were detected using multicolor flow cytometry reflecting population expansion after CMV infection. The presence of CMV DNA in saliva and plasma associated with plasma levels of antibodies reactive with CMV gB and with populations of circulating Vδ2− γδ T -cells (p < 0.01). T-cells reactive to CMV immediate early (IE)-1 protein were generally lower in patients with CMV DNA in saliva or plasma, but the level of significance varied (p = 0.02–0.16). Additionally, CMV DNA in saliva or plasma associated weakly with impaired FMD (p = 0.06–0.09). The data suggest that CMV detected in saliva reflects systemic infections in adult RTR.

Pulmonary cytomegalovirus (CMV) DNA shedding in allogeneic hematopoietic stem cell transplant recipients: Implications for the diagnosis of CMV pneumonia

To date no definitive cut-off value for cytomegalovirus (CMV) DNA load in bronchoalveolar lavage (BAL) fluid specimens has been established to discriminate between CMV pneumonia and pulmonary CMV DNA shedding in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients. The current retrospective study is aimed at assessing the range of CMV DNA loads quantified in BAL fluid specimens from allo-HSCT patients with pneumonia in which different microorganisms were causally involved. A total of 144 BAL specimens from 123 patients were included. CMV DNA was detected in 56 out of 144 BAL fluid specimens and the median CMV DNA load from patients in whom CMV pneumonia was unlikely or could be tentatively ruled out was 1210 (31-68, 920)IU/ml. The frequency of CMV DNA detection and median CMV DNA loads were comparable, irrespective of the attributable cause of pneumonia. Detection of CMV DNA loads in BAL fluid specimens >500IU/ml was independently associated with pneumonia-attributable mortality. The current study highlights the difficulty in establishing universal CMV DNA load thresholds in BAL fluid specimens for distinguishing between CMV pneumonia and pulmonary CMV DNA shedding, and suggests that the presence of CMV DNA in BAL fluid specimens beyond a certain level may have a deleterious impact on patient outcome.

Investigation of congenital CMV infection with the presence of CMV DNA in saliva samples of new born babies

Cytomegalovirus (CMV), is the most common cause among congenital infections and is the most seen etiology in long-term sensorineural hearing loss (SNHL) and neurological impairment. Congenital CMV infection (CCMV) was reported in 0.15-2.2% of live-borne neonates in studies from different countries. A significant proportion of infected infants are asymptomatic after birth and might only be detected by routine screening methods during the new born period. The aim of this study was to screen the saliva of live-born neonates with areal-time PCR based method for the detection of CCMV in our hospital. Saliva samples collected in half an hour after birth by dry dacron swabs and were evaluated for CMV DNA (Rt-PCR, Abbott Molecular USA) from 1000 babies born in Ege University Faculty of Medicine Hospital Obstetrics Clinic between October 2015-October 2017. For the confirmation of CCMV, saliva positive newborns were evaluated with the same method for CMV DNA from their urine or blood within 21 days. All newborns were screened for sensorineural hearing tests. Subjects were 497 girls (49.7%) and 503 boys (50.3%), with a mean weight of 3116.8 g and mean of 37.61 birth week. CMV DNA was positive in the saliva of 16 newborns (1.6%). Fourteen newborns were weakly positive for CMV DNA in their saliva and were not confirmed for CCMV infection. Congenital CMV was confirmed in only two (0.2%) with the CMV DNA results in urine and/or blood samples. One of the two newborns with CCMV was symptomatic and had a neurosensorial hearing loss. The other one was asymptomatic. Saliva samples, taken immediately after birth with a noninvasive and easy method for the detection of CMV DNA is very important for diagnosis of CCMV. Positive samples should be confirmed with CMV DNA in urine or blood samples of these newborns. In this study, detection of positivity in saliva samples that were confirmed with other samples of our newborn population for CCMV was 0.2%. The specific diagnosis for CCMV in newborns with a noninvasive and easy collecting sample is important to avoid sequelae and for public health concerns.

Evaluation of chronic inflammation in the aetiology of venous insufficiency by investigating cytomegalovirus DNA.

IntroductionLower extremity venous insufficiency is a significant health problem with economic and sociological consequences, lowering the quality of life, and sometimes leading to serious complications. The aim of this study is to evaluate the cytomegalovirus (CMV) effect on chronic inflammation in the aetiology of chronic venous insufficiency.Material and methodsBetween November 2017 and August 2018, 468 patients who underwent radio-frequency ablation therapy and phlebectomy were included in the study. PCR analyses for CMV DNA were performed on the venous tissue samples. Patients with post-thrombotic syndrome were excluded from the study. After ethical approval, the relationship between the presence of CMV DNA, gender, body mass index, and bilaterality of chronic venous insufficiency were investigated.ResultsWhen the relationship between CMV DNA and gender or body mass index was examined, a significant relationship was not detected. But when the patients with bilateral chronic venous insufficiency and patients with unilateral chronic venous insufficiency were compared regarding CMV DNA positivity, the patients with bilateral chronic venous insufficiency had significantly higher CMV DNA positivity (p = 0.002). Also, the incidence of venous ulcers in the CMV DNA exposed group was significantly higher.ConclusionsIn the literature there are many studies showing that CMV triggers atherosclerosis, but there is no study in which CMV directly produces chronic venous insufficiency. The high rates of positivity suggest that CMV, which is the basis of chronic inflammation, may be a significant factor in the aetiology of chronic venous insufficiency.

Прогностическая значимость уровня противовирусных антител в развитии рецидива рака мочевого пузыря

Serological diagnosis of virus-associated tumors attracts the attention of many specialists. The changes in the level of antiviral antibodies in tumors of different localizations are proved. In some cases, the authors suggest using these data either for screening of tumors or for controlling the cure. to evaluate the predictive value of antiviral antibodies for the recurrence of bladder cancer. a level of antiviral antibodies (IgG, M) against Epstein-Barr virus (EBV), herpes simplex virus (HSV) 1 and 2 types, cytomegalovirus (CMV) of 100 patients with bladder cancer (72 men and 28 women) aged from 38 to 90 years (mean age 65+/-10) was studied. Multivariate analysis with a construction of classification tree was performed. The recurrence of the bladder cancer was used as the dependent variable. in patients with recurrence of bladder cancer there was an increase in the level of anti-CMV IgG (616.5+/-501.46 U/ml vs. 339.06+/-306.61 U/ml, p=0.0017) and anti-EBV IgG-EBNA (246,7+/-207 U/ml vs. 141,5+/-163,7 U/ml, p=0.0118). After the construction of the classification tree, anti-CMV IgG, anti-EBV IgG-EBNA, tumor stage and the presence of CMV DNA in tumor tissue were selected. It allowed to classify correctly 20 of 24 patients with recurrence and 58 of 72 patients without relapse. The most significant predictors included anti-CMV IgG level (100%), anti-BNA IgG level (78%) and tumor stage (50%). The sensitivity, specificity, positive prognostic value (probability of tumor detection in patients with a positive test result), negative prognostic value (probability of absence of the tumor in persons with a negative test result) and accuracy were 83.33%, 80.56%, 58.82%, 93.55% and 81.25%, respectively. A multivariate analysis (binary logistic regression) was performed and a reliable model (2=22,438, p=0,00043) was created, including the following parameters: anti-CMV IgG more than 670 u/ml, anti-BNA IgG more than 130 u/ml, the degree of anaplasia, the presence CMV and/or EBV DNA in tumor tissue. Based on the regression equation, an accuracy test for prediction of tumor recurrence was carried out, which resulted in fairly high predictive results: specificity and sensitivity were 95.2% and 33.3%, respectively. anti-CMV IgG level more than 670 U/ml and anti-BNA IgG level more than 130 U/ml are reliable predictors for the recurrence of bladder cancer.

Maternal and neonatal screening methods for congenital cytomegalovirus infection.

Human cytomegalovirus (CMV) is a common cause of congenital infection that may lead to severe long-term sequelae. Because there are no established vaccines, fetal interventions or neonatal treatments, neither maternal nor neonatal screening is recommended. However, recent studies have indicated that early antiviral treatment may improve neurological outcomes in symptomatic infants with congenital infection. Therefore, prenatal detection may be important in newborns at high risk of such infection. Polymerase chain reaction for CMV DNA in the amniotic fluid is considered the gold standard for diagnosis of intrauterine infection, but its use is limited because amniocentesis is an invasive procedure. In a prospective cohort study, we have reported that the presence of CMV DNA in secretions of the maternal uterine cervix were predictive of congenital infection in groups at high risk. However, we also recently demonstrated that maternal serological screening for primary CMV infection using specific immunoglobulin G, the immunoglobulin G avidity index or specific immunoglobulin M can overlook many cases. Previous research has indicated that the combination of early detection by universal neonatal screening of urinary CMV DNA combined with early antiviral therapy can improve outcomes in infants with symptomatic congenital infection. In this article, we review the current state of maternal and neonatal screening for congenital CMV infection.

18F-FDG PET/CT Findings in Cytomegalovirus Colitis.

We present a case demonstrating the diagnostic work-up of a patient undergoing azathioprine treatment for inflammatory bowel disease (IBD), diagnosed with an acute cytomegalovirus (CMV) infection and CMV colitis. An 18F-FDG positron emission tomography/computed tomography (PET/CT) performed 2 weeks after debut of symptoms revealed pathological 18F-FDG uptake in the left side of the colon mucosa, mimicked activity of IBD. However, a diagnosis of CMV colitis was based on the presence of CMV IgM antibodies, a seroconversion of CMV IgG antibodies, presence of CMV DNA in plasma and the finding af CMV DNA in biopsies from the intestinal mucosa. The patient responded to treatment with ganciclovir. This case highlights that a positive 18F-FDG PET/CT scan of the colon can be due to CMV colitis.

Characterization of Natural Killer Cells in HIV Patients Beginning Therapy with a High Burden of Cytomegalovirus

ABSTRACTBackground: Active infections with cytomegalovirus (CMV) increase NK cell expression of the inhibitory receptor LIR-1 and the activating receptor NKG2C in transplant recipients. However, the effects of CMV on NK cells are different in HIV patients stable on antiretroviral therapy (ART) and have not been analyzed in young HIV patients beginning ART.Methodology: We followed a cohort of 78 Indonesian HIV patients beginning ART. CMV antibodies were measured in plasma before ART (baseline), and after 1, 3, 6, and 12 months. CMV DNA was sought in blood granulocytes at baseline by quantitative PCR assay and a deletion in the NKG2C gene was identified by PCR. NK cell profiles were monitored by flow cytometry in 19 patients stratified by the presence of CMV DNA. Healthy controls (n = 17) were assessed once.Results: All 78 patients were CMV seropositive and 41 had detectable CMV DNA. CMV DNA+ patients had higher proportions of total NK cells and CD16+ NK cells at baseline, but similar expression of LIR-1 and NKp30 on NK cells on ART. However, levels of CMV antibody were inversely related to median LIR-1 expression on NK cells. A dramatic elevation in cells expressing NKG2C was restricted to CMV DNA+ patients heterozygous for the NKG2C deletion. Patients with High NKG2C expression had lower levels of CMV antibodies.Conclusion: A subpopulation of NK cells expressing NKG2C was induced by CMV replication in HIV patients heterozygous for a deletion in this gene. Individuals with an abundant NKG2C+ and LIR-1+ NK cells displayed lower levels of CMV reactive antibody.

Associations between patient clinical characteristics and the presence of cytomegalovirus DNA in the bronchoalveolar lavage fluid of children with recurrent wheezing

BackgroundThis study aimed to investigate the occurrence of Cytomegalovirus (CMV) DNA in the Bronchoalveolar lavage fluid (BALF) of children with recurrent wheezing and to identify associations with certain patient clinical characteristics.MethodsIn this cross-sectional study, pediatric patients (age < 36 months) admitted to Soochow University Hospital with recurrent wheezing (≥ 4 episodes of wheezing per year) were enrolled in the study. Cytomegalovirus DNA from their BALF was detected by real-time PCR. Subpopulations of blood immunoglobulins and T lymphocytes were quantified. The clinical characteristics of patients with and without BALF CMV DNA were compared. Comparisons of non-normally distributed continuous variables between groups were made using the Mann-Whitney U-test. Comparisons of frequency distributions were made using the Chi-squared test. Spearman’s rank correlation coefficient was used to evaluate correlations between the number of CMV DNA copies and continuous variables.ResultsA total of 111 patients aged 4 to 36 months (median 14.0 (IQR 8.0–22.0) months) were enrolled on to the study. Cytomegalovirus DNA was detected in 51.4% of patients (n = 111) with recurrent wheeze and was more prevalent among those aged 12 to 36 months with a positive modified asthma predictive index (mAPI) (n = 38, median 23.5 (IQR 19.7–31.2) months) than in those of the same age group with a negative mAPI (n = 25, median 15.0 (IQR 13.0–19.0) months) (57.9% vs. 20.0%, p = 0.003). Bronchoalveolar lavage fluid CMV DNA copy number [median 7560 (IQR 1200–71,150) copies/mL] was positively correlated with the duration of hospitalization (r = 0.33, p = 0.013), and negatively correlated with patient age (r = − 0.41, p = 0.002) and the percentage of BALF eosinophils (r = − 0.38, p = 0.004).ConclusionsCMV infection or reactivation in children with recurrent wheeze is associated with certain clinical characteristics, including younger age and lower levels of BALF eosinophils. Higher CMV DNA copy numbers were associated with a longer duration of hospitalization. Further studies are needed to address whether specific antiviral treatment could be beneficial for BALF CMV positive patients.

CMV on surfaces in homes with young children: results of PCR and viral culture testing

BackgroundCaring for young children is a known risk factor for cytomegalovirus (CMV) infection mainly through exposure to their saliva and urine. In a previous study, 36 CMV-seropositive children 2 mo. to 4 years old were categorized as CMV shedders (n = 23) or non-shedders (n = 13) based on detection of CMV DNA in their saliva and urine. The current study evaluated the presence of CMV on surfaces in homes of the children.MethodsStudy staff made 4 visits to homes of the 36 enrolled children over 100 days. Saliva was collected by swabbing the mouth and urine was collected on filter paper inserted into diapers. In addition, five surface specimens were collected: three in contact with children’s saliva (spoon, child’s cheek, washcloth) and two in contact with children’s urine (diaper changing table, mother’s hand). Samples were tested by PCR and viral culture to quantify the presence of CMV DNA and viable virus.ResultsA total of 654 surface samples from 36 homes were tested; 136 were CMV DNA positive, 122 of which (90%) were in homes of the children shedding CMV (p < 0.001). Saliva–associated samples were more often CMV positive with higher viral loads than urine-associated samples. The higher the CMV viral load of the child in the home, the more home surfaces that were PCR positive (p = 0.01) and viral culture positive (p = 0.05).ConclusionsThe main source for CMV on surfaces in homes was saliva from the child in the home. Higher CMV viral loads shed by children correlated with more viable virus on surfaces which could potentially contribute to viral transmission.

Cytomegalovirus infection alters phenotypes of different γδ T‐cell subsets in renal transplant recipients with long‐term stable graft function

Cytomegalovirus (CMV) infection alters the phenotypic profiles of T-cells and NK cells in healthy and immunocompromised individuals. Here, we examined the effects of CMV infection on the phenotype and functions of γδ T-cell subsets in renal transplant recipients (RTR) stable several years after transplantation (n = 80) and healthy controls (n = 72). Differentiation status, function, and expression of HLA-DR, CD57, and LIR-1 on Vδ2- and Vδ2+ γδ T-cells were examined in peripheral blood cells using flow cytometry. Percentages of Vδ2- γδ T-cells were higher in RTR who are CMV-seropositive and correlated with CMV antibody levels. Proportions of Vδ2- γδ T-cells expressing HLA-DR, CD57, or LIR-1 were increased in CMV-seropositive RTR and healthy controls compared to their seronegative counterparts. Additionally, Vδ2- γδ T-cells were skewed towards a terminally differentiated phenotype and most expressed CD8 in individuals who were CMV-seropositive. Increased expression of LIR-1 on terminally differentiated Vδ2- γδ T-cells was associated with CMV seropositivity in RTR and controls. The presence of CMV DNA in 15 RTR was associated with higher frequencies of LIR-1+ Vδ2+ γδ T-cells and increased percentages of terminally differentiated effector memory cells in both γδ T-cell subsets. Our study further characterises the effects of CMV and transplantation on γδ T-cell phenotypes.

Presence of Cytomegalovirus DNA in the Whole Blood Is Associated with the Risk of Bronchial Asthma

Presence of Cytomegalovirus DNA in the Whole Blood Is Associated with the Risk of Bronchial Asthma

ATPS-33EVIDENCE OF ABSENCE FOR CYTOMEGALOVIRUS IN PATIENTS WITH GLIOBLASTOMAS AND OTHER HIGH-GRADE GLIOMAS BY REAL-TIME PCR FOR DNA, IMMUNOHISTOCHEMISTRY FOR PROTEIN ANDIN-SITUHYBRIDIZATION FOR mRNA

BACKGROUND: Several groups have suggested a potential causative role of cytomegalovirus (CMV) in the development of high-grade gliomas (HGG), while others have not found any evidence of CMV in these cancers. We used 3 different molecular technologies to assay for the presence of CMV DNA, mRNA and protein in tumor tissue and blood samples of patients with HGG. METHODS: Part A: Fresh-frozen tissues from 27 glioblastomas (GBM) were analyzed for CMV by real-time PCR of US17. Part B: Prospective study of patients with newly-diagnosed HGG (grades III or IV) who underwent standard chemoradiation. Plasma and serum were collected at baseline, and plasma at follow-up visits. FFPE tumor tissue was analyzed for CMV proteins by genetically validated immunohistochemistry (IHC), in-situ hybridization assays targeting 1E1/2 and pp65, and by real-time PCR for US17 (tissue and plasma). RESULTS: Part A: CMV was detected in 0/27 fresh-frozen GBM tissue samples using real-time PCR. Part B: Eighteen patients with HGG (12 grade IV, 6 grade III) were enrolled. CMV tissue testing was negative in all samples by both probes for in-situ hybridization, both antibodies for IHC (n = 11 available samples), and CMV US17 real-time PCR (PCR successful in 8/14 samples). Serum CMV IgG avidity index was positive (0.55-0.82) in 8/15 cases, consistent with rates in the general population. CMV was detected in baseline plasma samples of 3/16 patients and in 1 single follow-up sample. All other plasma samples were negative for CMV (n = 34). CONCLUSION: CMV was not detected in any HGG tissue. Half of the patients with HGG did not have an exposure to CMV prior to their diagnosis, and viremia was rare at presentation and follow-up. An independent, unbiased investigation of CMV positivity rates using a central reference laboratory with standardized tissue samples and controls is warranted before further consideration of CMV-targeted therapy in HGG.

Investigation of presence of Cytomegalovirus DNA by real-time polymerase chain reaction in transplant recipients

Investigation of presence of Cytomegalovirus DNA by real-time polymerase chain reaction in transplant recipients

G172(P) Case report:cytomegalovirus and kawasaki disease – is there a link?

Aims Kawasaki disease is an uncommon cause of fever in a young infant (less than 6 months) and is associated with life-threatening complications. Cytomegalovirus has been linked with atherosclerosis of coronary arteries in some adult studies. Acute Cytomegalovirus infection has been detected in a few reported cases of Kawasaki disease in infants. Methods A retrospective case review was done of a 3 month old Asian infant, who presented to a district general hospital, with a 3 day history of fever and irritability. She was treated with intravenous antibiotics for presumed sepsis. The fever persisted after 5 days and she was transferred to a tertiary hospital for further investigations into the cause of fever. Results Cytomegalovirus DNA was detected by polymerase chain reaction (PCR) in the infant’s blood. Cytomegalovirus IgM was not detected. ECG and echocardiogram were not performed due to lack of clinical evidence of underlying cardiac cause at the time. The fever was settling and infection markers were improving. The infant looked to be improving, according to the parents. The infant became acutely unwell, arrested and passed away on day 13 of illness. At post-mortem, multiple thrombosed giant aneurysms (up to 17mm) of the coronary arteries were found. There were multiple saccular coronary artery aneurysms affecting the right coronary (11 × 17mm), left descending coronary artery, left diagonal artery, left circumflex and the right posterior descending artery. The pericardial cavity contained 70mls of blood. There was moderate to severe coronary artery arteritis. The cause of death was cardiac tamponade secondary to rupture of a coronary artery aneurysm, caused by Kawasaki disease. Cytomegalovirus DNA was detected in liver and lung tissue. Conclusion Kawasaki disease is an uncommon but important differential in a young infant with fever and requires prompt treatment with immunoglobulins and Aspirin. If echocardiography is not available, an ECG may provide useful information such as ST segment changes and Q waves. The presence of Cytomegalovirus DNA on blood PCR raises the possibility of Kawasaki disease and a positive result may be helpful additional information in the diagnosis of Kawasaki disease.

No evidence of association between human cytomegalovirus infection and papillary thyroid cancer

BackgroundHuman cytomegalovirus (CMV) has been detected in the thyroid gland and thyroid tumors. CMV infection may activate the mitogen-activated protein kinase pathway, of which aberrant activation is frequently associated with BRAF mutation in papillary thyroid cancer.MethodsA total of 45 paired tumorous and adjacent non-neoplastic tissue samples, including 5 follicular adenoma and 40 papillary thyroid cancer, were obtained during thyroidectomy. BRAF mutational status was determined using direct sequencing. The presence of CMV DNA was determined using conventional PCR and quantitative real-time PCR. CMV protein in the tissue samples were evaluated with Western blot analysis.ResultsBRAF mutation was identified in the cancerous part of 31 (78%) papillary thyroid cancers. Papillary cancer with BRAF mutation was significantly associated with a larger tumor size (P = 0.045), extrathyroidal invasion (P = 0.012), lymph node metastasis (P = 0.008), and a higher TNM stage (P = 0.044). CMV DNA and protein were not detected in any studied samples.ConclusionsOur results suggest no association between CMV infection and papillary thyroid cancer.