Assessment of blood viral load in asymptomatic and symptomatic cases of congenital cytomegalovirus infection.

This study aimed to retrospectively evaluate the baseline and follow-up viral loads and viral clearance times in cases followed for asymptomatic and symptomatic congenital cytomegalovirus (cCMV) infection between August 2010 and August 2022. Among 93 cases, they had asymptomatic (n: 55) and symptomatic (n: 38). The median baseline blood viral load detected in the symptomatic cCMV (ScCMV) infection (13 054 IU/mL) was significantly higher than that of asymptomatic cCMV (AcCMV) infection (4636 IU/mL) (p < 0.013). There was no difference in median viral clearance times (75 and 90 days, respectively) in baseline viremic cases in the ScCMV and AcCMV infection groups. There were no differences in median baseline blood viral load (6930 IU/mL and 14 268 IU/mL, respectively) and median viral clearance times (75 and 85 days, respectively) between the 6-week and 6-month antiviral treatment group. No correlation was found between baseline blood viral load, clinical severity, and the number of systems involved. However, in initial viremic cases, the viral load threshold for a symptomatic case was 8856 IU/mL, with 85.7% sensitivity and 54.5% specificity.

TTV and CMV viral load dynamics: Which emerges first during immunosuppression?

Novel biomarkers reflecting the degree of immunosuppression in transplant patients are required to ensure eventual personalized equilibrium between rejection and infection risks. With the above aim, Torque Teno Virus (TTV) viremia was precisely examined in a large cohort of transplanted immunocompromised patients (192 hematological and 60 solid organ transplant recipients) being monitored for Cytomegalovirus reactivation. TTV load was measured in 2612 plasma samples from 448 patients. The results revealed a significant increase in TTV viral load approximately 14 days following CMV reactivation/infection in solid organ transplant (SOT) patients. No recognizable difference in TTV load was noted among hematological patients during the entire timeframe analyzed. Furthermore, a temporal gap of approximately 30 days was noted between the viral load peaks reached by the two viruses, with Cytomegalovirus (CMV) preceding TTV. It was not possible to establish a correlation between CMV reactivation/infection and TTV viremia in hematological patients. On the other hand, the SOT patient cohort allowed us to analyze viral kinetics and draw intriguing conclusions. Taken together, the data suggest, to our knowledge for the first time, that CMV infection itself could potentially cause an increase in TTV load in the peripheral blood of patients undergoing immunosuppressive therapy.

Assessment of the impact of cytomegalovirus seropositivity on blood parameters in renal hemodialysis patients.

End-stage renal disease (ESRD) patients are considered immunocompromised, putting them at high risk for infections, including cytomegalovirus (CMV). CMV can affect hematological parameters, causing further complications in ESRD patients. This study intended to determine the seropositivity of CMV infection in hemodialysis patients and its effect on different blood parameters in ESRD patients to help decrease the overall dialysis associated morbidity and mortality. Blood samples were collected from 45 ESRD patients and 45 controls. A complete blood count was performed using an automated cell counter. CMV-specific IgM and IgG levels were measured using immunochemistry testing. The seropositivity for CMV-IgG was 42.2% in ESRD patients which was significantly higher than in control group (22.2%) (p=0.042). The seropositivity for CMV-IgM was 6.7% in ESRD patients with no difference with the control group (4.4%). The prevalence of anemia was significantly higher in CMV seropositive (77.3%) compared to CMV seronegative (47.8%) ESRD patients. Other studied blood parameters were not different between CMV seronegative and seropositive ESRD patients. In conclusion, CMV infection is a significant concern for dialysis patients and can affect hematological parameters, leading to further complications. Early detection and treatment of CMV infection and monitoring of CMV IgM and IgG levels are critical to prevent further complications and improve clinical outcomes.

Cytomegalovirus antigen-specific multi-cytokine immune responses in patients with rheumatic diseases under different cytomegalovirus infection status: A case-control study.

OBJECTIVE: To explore Cytomegalovirus (CMV) antigen-specific multi-cytokine immune responses in patients with rheumatic disease (RD) under different CMV infection status. METHODS: A total of 60 RD patients in our center from March 2023 to August 2023 were enrolled. The patients were divided into latent CMV infection and active CMV infection, the latter was classified as subclinical CMV infection or CMV disease based on presence or absence of symptoms related to CMV. Whole blood was collected and stimulated with QuantiFERON-CMV antigen. The levels of IFN-γ, TNF-α, IL-2, IL-4, IL-6, IL-10, IL-17 and CXCL-2 in supernatant were measured by Luminex Assays. The receiver operating characteristic curve was used to evaluate the diagnostic accuracy of cytokine for distinguishing different CMV infection status. RESULTS: The proportion of patients with severe lymphopenia was lowest in the latent CMV infection group, while there were no significant differences in medication usage in different CMV infection status. After stimulation with QF-CMV antigens, the levels of IFN-γ, TNF-α and IL-2 in the CMV disease group were significantly lower than those in the latent CMV infection group. CMV antigen-specific IFN-γ, TNF-α levels and severe lymphopenia together provided the best discriminatory performance for distinguishing between latent and either active CMV infection patients (AUC = 0.854) or CMV disease patients (AUC = 0.935). CONCLUSION: Noninvasive peripheral blood biomarkers (the combination of CMV antigen-specific IFN-γ, TNF-α levels and severe lymphopenia) may have the potential to diferentiate different status of CMV infection in RD population.

Comparison of Whole Blood and Plasma for Monitoring Cytomegalovirus and Epstein-Barr Virus.

Monitoring Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infection after transplantation is recommended to enable preemptive therapy. However, the most suitable sample type remains unclear. Patients who underwent hematopoietic stem cell or liver transplantation were included in this study. Viral loads in sequential whole-blood and plasma samples were retrospectively analyzed. EBV DNA was detected more frequently in whole blood (55%) than in plasma (18%). The detection rate of CMV DNA was similar between the two sample types. The correlation of viral loads between the two sample types were 0.515 and 0.688 for EBV and CMV, respectively. Among paired samples in which EBV DNA was detected in whole blood, the plasma EBV detection rate was significantly higher in patients who underwent hematopoietic stem cell transplantation than in those who underwent liver transplantation. The viral DNA load in whole blood and plasma showed similar trends. The EBV detection rate was higher in whole blood, and a high correlation was observed between CMV DNA loads and whole blood and plasma. These results indicate that whole blood is more sensitive for monitoring both EBV and CMV, whereas plasma is a potential alternative sample for monitoring CMV.

Evaluation of the Tasso+ blood self-collection device for quantitation of plasma cytomegalovirus (CMV) DNAemia in adult solid organ transplant recipients (SOTr).

Quantitative monitoring of cytomegalovirus (CMV) DNAemia in venous blood is standard in solid organ transplant recipients (SOTr) but is limited by the need for phlebotomy facilities and personnel. The aim of the study was to evaluate the Tasso+ capillary blood (CB) self-collection device for quantitation of plasma CMV DNAemia. Thirty adult SOTr with suspected CMV DNAemia were enrolled to have a supervised Tasso+ CB sample collection within 24 h of a venous sample. CMV DNA was quantitated in paired samples by using the Abbott M2000 Real-Time qPCR instrument. The participants were provided with a study-specific survey that measured patient acceptability of the Tasso+ device compared with venipuncture. A Tasso + CB sample was successfully collected in 28/30 (93%) patients, and 44 paired samples were analyzed. Concordance for detection of CMV DNAemia above the limit of detection (LOD) was 91% (42/44), and the Tasso + CB sample was estimated to be 95% sensitive at a viral load (VL) of 308 IU/mL. Among samples with a quantifiable DNAemia result with both methods (N = 31), there was excellent correlation between methods (Spearman R2 = 0.99). The difference in CMV VL between venous and Tasso+ CB samples was not dependent on time (P > 0.1). Of 12 who completed the survey, 11 (92%) expressed a preference for Tasso+ CB collection over venipuncture. Collection of CB with the Tasso+ device is feasible, patient-acceptable, and yields generally comparable CMV DNAemia load to standard venous samples, but with lower sensitivity. Future studies to optimize and evaluate this methodology for patient self-collected samples are warranted. IMPORTANCE: We evaluate an FDA-cleared blood self-collection device (Tasso+) and demonstrate that it is patient-acceptable and yields a liquid blood sample with quantitative CMV DNAemia results comparable to those of standard venipuncture samples. This opens up possibilities for self-blood collection to monitor for CMV and potentially other viruses in transplant and other at-risk populations.

Analytical and Clinical Performance of the NeuMoDx™ Platform for Cytomegalovirus and Epstein-Barr Virus Viral Load Testing.

DNA assays for viral load (VL) monitoring are key tools in the management of immunocompromised patients with cytomegalovirus (CMV) or Epstein-Barr virus (EBV) infection. In this study, the analytical and clinical performances of the NeuMoDx™ CMV and EBV Quant Assays were compared with artus CMV and EBV QS-RGQ Kits in a primary hospital testing laboratory. Patient plasma samples previously tested using artus kits were randomly selected for testing by NeuMoDx assays. The NeuMoDx CMV Quant Assay and artus CMV QS-RGQ Kit limits of detection (LoDs) are 20.0 IU/mL and 69.7 IU/mL, respectively; 33/75 (44.0%) samples had CMV DNA levels above the LoD of both assays. The Pearson correlation coefficient was 0.9503; 20 samples (60.6%) had lower NeuMoDx CMV quantification values versus the artus kit. The LoD of the NeuMoDx EBV Quant Assay and artus EBV QS-RGQ Kit are 200 IU/mL and 22.29 IU/mL, respectively; 16/75 (21.3%) samples had EBV DNA levels above the LoD of both assays. The Pearson correlation coefficient was 0.8990. EBV quantification values with the NeuMoDx assay were higher versus the artus kit in 15 samples (93.8%). In conclusion, NeuMoDx CMV and EBV Quant Assays are sensitive and accurate tools for CMV and EBV DNA VL quantification.

High-dimensional mass cytometry identified circulating natural killer T-cell subsets associated with protection from cytomegalovirus infection in kidney transplant recipients.

Cytomegalovirus (CMV) infection is associated with poor kidney transplant outcomes. While innate and adaptive immune cells have been implicated in its prevention, an in-depth characterization of the in vivo kinetics of multiple cell subsets and their role in protecting against CMV infection has not been achieved. Here, we performed high-dimensional immune phenotyping by mass cytometry, and functional assays, on 112 serially collected samples from CMV seropositive kidney transplant recipients. Advanced unsupervised deep learning analysis was used to assess immune cell populations that significantly correlated with prevention against CMV infection and anti-viral immune function. Prior to infection, kidney transplant recipients who developed CMV infection showed significantly lower CMV-specific cell-mediated immune (CMI) frequencies than those that did not. A broad diversity of circulating cell subsets within innate and adaptive immune compartments were associated with CMV infection or protective CMV-specific CMI. While percentages of CMV (tetramer-stained)-specific T cells associated with high CMI responses and clinical protection, circulating CD3+CD8midCD56+ NK-T cells overall strongly associated with low CMI and subsequent infection. However, three NK-T cell subsets sharing the CD11b surface marker associated with CMV protection and correlated with strong anti-viral CMI frequencies in vitro. These data were validated in two external independent cohorts of kidney transplant recipients. Thus, we newly describe the kinetics of a novel NK-T cell subset that may have a protective role in post-transplantation CMV infection. Our findings pave the way to more mechanistic studies aimed at understanding the function of these cells in protection against CMV infection.

Evaluation of Toxoplasma, Rubella, and Cytomegalovirus serological results in women of childbearing age

SUMMARY OBJECTIVE This study aimed to determine the rates of IgG and IgM antibodies against cytomegalovirus, rubella, and Toxoplasma gondii (all of which may cause congenital infections) in women of childbearing age who were admitted to Bolu Abant İzzet Baysal University Training and Research Hospital. METHODS Between January 2015 and December 2017, Toxoplasma gondii, rubella, and cytomegalovirus IgM and IgG antibody levels were studied using the ELISA method (Architect i2000SR, Abbott, Germany) in patients aged 15 to 45 who attended the obstetrics and gynecology outpatient clinics. Toxoplasma gondii and cytomegalovirus IgG avidity levels were analyzed retrospectively. RESULTS A total of 13.470 tests were conducted in the laboratory. Seropositivity percentages of IgM antibodies were found to be 1.3%, 0.5%, and 1.6% for Toxoplasma (n = 3607), rubella (n = 3931), and cytomegalovirus (n = 3795), respectively. The seropositivity percentages of IgG antibodies were 22%, 94.2%, and 98.2% for Toxoplasma (n = 702), rubella (n = 693), and cytomegalovirus (n = 679), respectively. Primary infection (acute, recently acquired) was found in 7 (35%) patients with low Toxoplasma IgG avidity. One (3%) patient with low cytomegalovirus IgG avidity had a primary infection. CONCLUSION Toxoplasma gondii seronegativity was found to be high in the region. Therefore, screening women of childbearing age may be important for the prevention of congenital infections caused by Toxoplasma gondii.

RESUMO OBJETIVO O objetivo deste estudo foi determinar as taxas de anticorpos IgG e IgM contra citomegalovírus, rubéola e Toxoplasma gondii (todos os quais podem causar infecções congênitas) em mulheres em idade fértil que foram admitidas no Hospital de Pesquisa e Treinamento da Universidade Bolu Abant İzzet Baysal. MÉTODOS Entre janeiro de 2015 e dezembro de 2017, os níveis de anticorpos IgG e IgM para Toxoplasma gondii, rubéola e citomegalovírus foram estudados usando o método Elisa (Architect i2000SR, Abbott, Alemanha) em pacientes de 15 a 45 anos que compareceram a ambulatórios de obstetrícia e ginecologia. Os níveis de avidez de IgG para Toxoplasma gondii e citomegalovírus foram analisados retrospectivamente. RESULTADOS Um total de 13.470 testes foram realizados em laboratório. As porcentagens de soropositividade dos anticorpos IgM foram de 1,3%, 0,5% e 1,6% para Toxoplasma (n=3.607), rubéola (n=3.931) e citomegalovírus (n=3.795), respectivamente. As porcentagens de soropositividade dos anticorpos IgG foram 22%, 94,2% e 98,2% para Toxoplasma (n=702), rubéola (n=693) e citomegalovírus (n=679), respectivamente. Infecção primária (aguda, adquirida recentemente) foi encontrada em sete (35%) pacientes com baixa avidez para Toxoplasma IgG. Um (3%) paciente com baixa avidez para citomegalovírus IgG teve uma infecção primária. CONCLUSÃO A soronegatividade do Toxoplasma gondii foi alta na região. Portanto, testar mulheres em idade fértil pode ser importante para a prevenção de infecções congênitas causadas pelo Toxoplasma gondii.

Standardization of antigenemia and qpcr cut-off values in whole blood for the detection of cytomegalovirus disease in hiv patients

Abstract INTRODUCTION: We defined the cut-off values of the antigenemia and cytomegalovirus (CMV) DNA tests in HIV/AIDS patients to identify CMV disease. METHODS: A total of 97 samples from 68 patients with and without CMV disease were analyzed by viral DNA detection and antigenemia assay. RESULTS: Qualitative and quantitative results significantly differed between assays. The cut-off values for the antigenemia and qPCR assays were 1.5 positive cells/200,000 leukocytes and 3.715 log/mL, respectively. CONCLUSIONS: Antigenemia and qPCR are suitable for monitoring CMV disease in HIV patients, however, the threshold values should be determined within the centers where the patients are monitored.

Pp65 antigenemia and cytomegalovirus diagnosis in patients with lupus nephritis: report of a series / Antigenemia pp65 e diagnóstico de citomegalovírus em pacientes com nefrite lúpica: série de casos

ABSTRACT Introduction: In contrast to organ transplantation, few studies correlate the monitoring of pp65 antigenemia with a diagnosis of cytomegalovirus (CMV) in patients with systemic lupus erythematosus (SLE). Objective: To highlight the importance of CMV outside transplantation, we monitored pp65 antigenemia in a series of SLE patients. Methods: From March 2015 to March 2016, SLE patients presenting kidney involvement, fever, and an unclear infection at hospital admission were monitored through pp65 antigenemia. The pp65 antigenemia assay, revealed by immunofluorescence, was correlated with clinical and laboratory findings. Results: We included 19 patients with a suspected unclear infection. A positivity for pp65 antigenemia was found in seven patients (36.8%). The mean age was 33.5 ± 11.2 years, 16 (84%) were females, and 16 (84%) were black. Lymphopenia, anemia, and higher scores of SLEDAI were significantly more common in pp65-positive patients. Five patients received antiviral therapy with ganciclovir. Although receiving specific CMV treatment, one patient died because of suspected CMV disease. Conclusions: Pp65 antigenemia might be relevant in SLE patients, and studies with a greater number of patients are needed in order to establish sensitivity and specificity of pp65 antigenemia in different clinical contexts of SLE patients.

RESUMO Introdução: Diferentemente do transplante de órgãos, poucos estudos correlacionam o monitoramento da antigenemia pp65 com o diagnóstico de citomegalovírus (CMV) em pacientes com lúpus eritematoso sistêmico (LES). Objetivo: De modo a destacar a importância do CMV para além do transplante, monitorizamos a antigenemia pp65 em uma série de pacientes com LES. Métodos: De março de 2015 a março de 2016, pacientes com LES que apresentaram acometimento renal, febre e infecção indeterminada na internação foram monitorados através da antigenemia pp65. O ensaio de antigenemia, revelada por imunofluorescência, foi correlacionado com achado clínicos e laboratoriais. Resultados: Foram incluídos 19 pacientes com suspeita de infecção indeterminada. Positividade para antigenemia pp65 foi encontrada em sete pacientes (36,8%). A idade média foi de 33,5 ± 11,2 anos; 16 (84%) eram do sexo feminino e 16 (84%) eram negros. Linfopenia, anemia e escore de SLEDAI mais elevado foram significativamente mais comuns em pacientes pp65 positivos. Cinco pacientes receberam terapia antiviral com ganciclovir. Apesar de receber tratamento específico para CMV, um paciente com suspeita de doença por CMV veio a óbito. Conclusões: Antigenemia pp65 pode ser relevante em pacientes com LES, e estudos com maior número de pacientes são necessários para estabelecer a sensibilidade e a especificidade da antigenemia pp65 em diferentes contextos clínicos envolvendo pacientes com LES.

Evaluation of diagnostic tests for cytomegalovirus active infection in renal transplant recipients / Avaliação de métodos diagnósticos para infecção ativa por citomegalovírus em receptores de transplante renal

Abstract Introduction: Cytomegalovirus (CMV) infection is a main viral infection after kidney transplantation. The diagnostic methods currently employed are pp65 antigenemia and nucleic acid amplification by polymerase chain reaction (PCR) and aim at detecting viral replication. Objective: The goal of this study was to evaluate and compare by both methods the incidence of CMV active infection in kidney transplant patients and to establishthe best clinical-laboratory correlation. Methods: Thirty sequential kidney transplant recipients were enrolled in a single center prospective cohort study. Peripheral blood samples were drawn from day 15 until the 6th month after transplantation and tested for CMV replication by pp65 antigenemia and quantitative PCR assays (qPCR). Results: Two hundred forty samples were analyzed and the incidence of active infection was similar by both methods. Time elapsed to the first positive test was almost identical but more samples tested positive by qPCR than by antigenemia in a behavior that was almost evenly distributed overtime. Agreement between tests was observed in 217 samples (90.4%; kappa = 0.529; p < 0.001) and in 25 patients the tests were concordant (83.3%; kappa = 0.667; p < 0.001). The evaluation of the diagnostic parameters for CMV replication revealed higher sensitivity for the qPCR test (82.1%) against antigenemia (59.0%). Quantitative PCR was also slightly more accurate than antigenemia. Conclusion: Our data demonstrate that both methods are suitable and have almost equivalent accuracy for the detection of post-transplant cytomegalovirus replication. The choice for either test must take in consideration the demand, execution capability and cost-effectiveness at each institution.

Resumo Introdução: Citomegalovírus (CMV) é uma importante causa de infecção viral após o transplante renal. Os métodos diagnósticos presentemente utilizados são a antigenemia pp-65 e os métodos que utilizam a amplificação de ácidos nucléicos pela reação em cadeia da polimerase (PCR) e visam à detecção da replicação viral. Objetivo: O objetivo deste estudo foi avaliar e comparar a incidência de infecção ativa por CMV em pacientes transplantados renais pelos dois métodos e estabelecer a melhor correlação clínico-laboratorial. Métodos: Trinta pacientes transplantados renais seqüenciais em um único centro foram incluídos em um estudo de coorte prospectiva. Amostras de sangue periférico foram coletadas a partir do 15º dia até o 6º mês pós-transplante e avaliadas para replicação de CMV por Antigenemia pp-65 e PCR quantitativo (qPCR). Resultados: Foram analisadas 240 amostras e a incidência de infecção ativa foi similar pelos dois métodos. O tempo médio transcorrido desde o transplante até o primeiro teste com resultado positivo foi quase idêntico entretanto mais amostras tiveram resultado positivo por qPCR do que antigenemia, um comportamento que se manteve quase uniforme ao longo do tempo. Concordância entre os testes foi observada em 217 amostras (90,4%; kappa = 0,529; p < 0,001) e em 25 pacientes (83,3%; kappa = 0,667; p < 0,001). A avaliação dos parâmetros diagnósticos para replicação de CMV revelaram maior sensibilidade para qPCR (82,1%) contra antigenemia (59,0%). PCR quantitativo também foi levemente mais preciso do que antigenemia. Conclusão: Nossos dados demonstram que ambos os métodos são adequados e tem precisão quase equivalente para a detecção da replicação do CMV após o transplante renal. A escolha entre um ou outro deve levar em consideração a demanda, capacidade de execução e custo-efetividade em cada instituição.

Infección por citomegalovirus en receptores de trasplante alogénico de células progenitoras hematopoyéticas: terapia temprana basada en el estudio de la antigenemia y la carga viral en sangre / Cytomegalovirus infection in allogeneic hemapotopoietic stem-cell transplantation: early treatment based on the study of antigenemia assay and viral load determination in blood

La prevención de la enfermedad por CMV en los receptores de trasplante de células progenitoras hematopoyéticas se basa en la terapia temprana de la reactivación viral. La Antigenemia y el estudio de la carga viral por PCR son las dos técnicas diagnósticas actualmente vigentes. Se siguió una cohorte de 35 pacientes con estudios semanales con ambos métodos desde la recuperación de la neutropenia hasta el día 100 días postrasplante. Se inició tratamiento empírico con antivirales (Ganciclovir o Foscarnet) con un resultado positivo (antigenemia > 1 cel/200.000 o carga viral > 500 copias / ml) y se mantuvo 3-6 semanas. La serología previa fue positiva en R y D en 66% de los casos, en R o D en 20%, negativa en 3% y no evaluable en 11%. Se detectó infección por CMV en el 50% de los pacientes. En 15 ptes el diagnóstico fue por PCR, en 2 ambas pruebas fueron positivas y en uno solo la antigenemia. Un paciente presentó neumonía por CMV y falleció dentro de los 100 días de seguimiento. En 11,4% de los casos se detectó reactivación viral luego de los 100 días y dos ptes presentaron neumonía por CMV tardía que fue causa de muerte. Conclusión: Con los umbrales utilizados la carga viral precedió a la antigenemia en el diagnóstico de reactivación de CMV. La terapia temprana previene la enfermedad temprana por CMV en la mayoría de los casos pero la enfermedad tardía es un problema pendiente de resolución (AU)

Prevention of CMV disease in hematopoietic stem-cell transplantation recipients is based on the early management of viral reactivation. Antigenemia assay and PCR viral load detection are the current diagnostic techniques of choice. We followed a cohort of 35 patients with weekly studies using both methods from recovery from neutropenia to day 100 post-transplant. Empirical viral treatment (Ganciclovir or Foscarnet) was started after a positive result (antigenemia > 1 cell/200,000 or viral load > 500 copies / ml) and maintained for 3-6 weeks. Previous serology was positive in R and D in 66% of the cases, in R or D in 20%, negative in 3%, and not evaluable in 11%. CMV infection was detected in 50% of the patients. In 15 patients the diagnosis was made using PCR, in 2 both tests were positive, and in one only the antigenemia assay was positive. One patient presented with pneumonia due to CMV and died within the 100 days of follow-up. In 11.4% of the cases viral activation was detected after 100 days and two patients developed late pneumonia due to CMV and consequently died. Conclusion: Using these thresholds viral load detection preceded antigenemia assay in the diagnosis of CMV reactivation. Early treatment prevents early disease due to CMV in the majority of cases, but late disease remains a problem to be solved (AU)

Seroprevalencia contra agentes ToRCH en mujeres indígenas en edad fértil, estado Zulia, Venezuela / Prevalence of infectious agents in indigenous women of childbearing age in Venezuela

Introducción. El termino ToRCH comprende a los patógenos Toxoplasma gondii, virus de la rubéola, citomegalovirus y virus herpes simple 1 y 2. En mujeres embarazadas expuestas pueden ser causa de abortos y malformaciones congénitas en el neonato. Objetivo. Determinar la seroprevalencia de infección por los agentes causantes del síndrome ToRCH en mujeres en edad fértil de algunas comunidades indígenas yukpa de Venezuela. Materiales y métodos. En el año 2007 fueron seleccionadas 109 muestras de 151 mujeres, en edades comprendidas entre 14 y 40 años. La detección de anticuerpos se hizo por el método de inmunoensayo enzimático indirecto o ELISA de Smartest Diagnostics™. Resultados. El 85,5 % presentó anticuerpos contra T. gondii, el 95,4 % para rubéola, el 75,2 % para citomegalovirus y el 97,2 % para el virus herpes simple 1 y 2. Se observa que el 21,1 % y el 30,2 % presentaron relación entre la variable aborto y las infecciones por citomegalovirus y virus herpes simple 1 y 2, respectivamente. Conclusiones. Existe alta seroprevalencia de infecciones por los agentes causantes del síndrome ToRCH en mujeres en edad fértil de la etnia indígena yukpa. Las condiciones sanitarias precarias y el consumo de agua contaminada con ooquistes, favorecen la adquisición de la infección por T. gondii. El hacinamiento, el inicio a temprana de edad de la actividad sexual y el número de parejas, pueden incidir en la presencia de citomegalovirus y virus herpes simple 1 y 2.

Introduction. The ToRCH syndrome includes the following infectious pathogens: Toxoplasma gondii, rubella, cytomegalovirus and herpes simplex virus 1 and 2. In susceptible pregnant women, these pathogens can cause abortions and congenital malformation in the newborn babies. Objective. The seroprevalence of infection by ToRCH agents was determined in women of childbearing age in several Venezuelan Yukpa indigenous communities. Material and methods. In 2007, 109 samples were selected from 151 women with an age range of 14 to 40 years old. The determination of antibodies against ToRCH agents was carried out through the indirect enzyme immunoassay technique by ELISA´s technique of Smartest Diagnostics. Results. Of the 109 samples, 85.5% presented antibodies against T. gondii, 95.4% for rubella, 75.2% for cytomegalovirus and 97.2% for and herpes simplex virus 1 and 2. A relationship between abortion and infection by cytomegalovirus and herpes simplex virus 1and 2 was noted in 21.1% and 30.2% of women presented, respectively. Conclusions. The findings show a high prevalence of ToRCH agents in women in childbearing age in Yukpa indigenous communities in Venezuela. Poor sanitary conditions and consumption of water contaminated with oocysts may be an important way of transmission of T. gondii. Overcrowding in the communities, sexual activity at an early age and number of partners and may be related to the presence of cytomegalovirus and herpes simplex virus HSV-1 and 2.

Alta prevalencia de IgG anti citomegalovirus en 583 embarazos: Hospital Padre Hurtado / High prevalence of IgG anti cytomegalovirus in 583 pregnancies: Hospital Padre Hurtado

ANTECEDENTES: Citomegalovirus (CMV) es la infección congénita más frecuente, demostrado en el 1% de recién nacidos en países desarrollados. Es la primera causa de sordera y alteraciones del desarrollo neuro-lógico infantil. Recientes estudios han demostrado que la seropositividad no evita una reinfección materna ni la enfermedad congénita, por lo que la caracterización de la seroprevalencia permite saber si la infección congénita proviene mayoritariamente de primoinfección o de reinfección. OBJETIVOS: Conocer la seroprevalencia al parto en 583 mujeres beneficiarías del Hospital Padre Hurtado durante mayo y junio del 2006. MÉTODOS: Estudio prospectivo, observacional, en que se estudio la presencia de IgG anti CMV en sangre materna al parto. RESULTADOS: Se obtuvo una seroprevalencia de 95%, sin casos de infección sintomática al nacer. CONCLUSIÓN: La seroprevalencia es elevada, lo que sugiere que la reinfección sería la forma principal de infección congénita. Un estudio en recién nacidos con cultivos virales o PCR permitiría conocer la tasa de infección congénita real, y no un estudio basado en seroconversión pues omitiría todos los casos que reinfección, que serían mayoritarios.

BACKGROUND: Cytomegalovirus is the most frequent congenital infection, affecting 1% of the population in developed countries, and the leading cause of deafness and brain development abnormalities in children. Recent studies have demonstrated that seropositivity do not avoid reinfection and congenital disease. OBJECTIVE: To study the seroprevalence in 583 pregnant women at delivery at Padre Hurtado Hospital, during 2006. METHODS: Prospective, observational study, in which maternal blood at delivery was studied for the presence of anti CMV IgG. RESULTS: There was 95% seroprevalence, without any case of symptomatic infection. CONCLUSION: The high prevalence supports that most of the cases of congenital disease would occur in seropositive women, supporting that reinfection is the main way of neonatal compromise. This supports that a study with direct detection in liveborns would be suitable to reveal the impact of cytomegalovirus in our population and not that of seroconversión.

Prognostic markers of symptomatic congenital cytomegalovirus infection

The objective of this research was to identify maternal and fetal characteristics as prognostic markers of congenital cytomegalovirus (CMV) infection. This is a descriptive study of 13 cases of congenital CMV infection referred to Institute de Puericulture et Perinatologie de Paris (IPP) from January 2005 to October 2006. Amniotic fluid puncture was performed to research CMV polimerase chain reaction (PCR). Cordocentesis and cord blood samples at delivery were also analyzed to determinate fetal platelets count, GGT, ASAT, ALAT, CMV-DNA and IgM antibody. Variables of symptomatic and asymptomatic infants were then compared. Data were analyzed by SPSS - 15.0. Mean gestational age of amniocentesis was 24.6 weeks and there was no difference of mean viral load in amniotic fluid considering infant features. Mean gestational age of cordocentesis was 26.1 weeks. There were no statistical differences of fetal viral load, IgM, platelets, GGT, ASAT and ALAT analyzed at cordocentesis samples, but at delivery, mean values of IgM and ASAT of fetal blood were increased in symptomatic ones (p= 0.03 for both parameters). When considering groups with normal and abnormal parameters, ASAT of cordon samples was also increased in symptomatic infants (p= 0.02). Sensibility, specificity, positive and negative predictive value of fetal ultrasound anomalies to detect symptomatic infants were, respectively, 80 percent, 62.5 percent, 57.1 percent and 83.3 percent. Thus, identification of markers of CMV symptomatic infants should be aimed. Prenatal diagnosis, identification and follow up of congenital CMV infected infants are important to consider treatment for symptomatic infants, trying to avoid or reducing some possible sequels.

Impact of cytomegalovirus and grafts versus host disease on the dynamics of CD57+CD28-CD8+ T cells after bone marrow transplant

OBJECTIVES: The present study aimed to evaluate the dynamics of CD28 and CD57 expression in CD8+ T lymphocytes during cytomegalovirus viremia in bone marrow transplant recipients. METHODS: In a prospective study, blood samples were obtained once weekly once from 33 healthy volunteers and weekly from 33 patients. To evaluate the expression of CD57 and CD28 on CD8+ T lymphocytes, flow cytometry analysis was performed on blood samples for four months after bone marrow transplant, together with cytomegalovirus antigenemia assays. RESULTS: Compared to cytomegalovirus-seronegative healthy subjects, seropositive healthy subjects demonstrated a higher percentage of CD57+ and a lower percentage of CD28+ cells (p<0.05). A linear regression model demonstrated a continuous decrease in CD28+ expression and a continuous increase in CD57+ expression after bone marrow transplant. The occurrence of cytomegalovirus antigenemia was associated with a steep drop in the percentage of CD28+ cells (5.94 percent, p<0.01) and an increase in CD57+ lymphocytes (5.60 percent, p<0.01). This cytomegalovirus-dependent effect was for the most part concentrated in the allogeneic bone marrow transplant patients. The development of acute graft versus host disease, which occurred at an earlier time than antigenemia (day 26 vs. day 56 post- bone marrow transplant), also had an impact on the CD57+ subset, triggering an increase of 4.9 percent in CD57+ lymphocytes (p<0.05). CONCLUSION: We found continuous relative changes in the CD28+ and CD57+ subsets during the first 120 days post- bone marrow transplant, as part of immune system reconstitution and maturation. A clear correlation was observed between the expansion of the CD57+CD28-CD8+ T lymphocyte subpopulation and the occurrence of graft versus host disease and cytomegalovirus viremia.

Despistaje de infección por citomegalovirus en niños VIH positivos mediante PCR: relación con serología y evolución clínica / PCR screening for cytomegalovirus infection in HIV-positive children: relationship between serology and clinical evolution

En pacientes VIH positivos es fundamental diagnósticar infección por CMV. La relación entre serología, detección viral y evolución clínica no está plenamente establecida. Detectar la presencia de CMV por PCR en sangre periférica en pacientes pediátricos, sin síntomas de infección, VIH positivos; relacionar estos resultados con la serología y la evolución clínica durante un año de seguimiento. Criterios de inclusión: Niños menores de 12 años, ambos sexos, infección diagnosticada por VIH, recibiendo terapia antirretroviral altamente efectiva (TARAE), consentimiento informado por escrito, firmado. La serología anti CMV se realizó mediante el método ELISA, la semicuantificación de CMV en sangre periférica, mediante PCR, y el inmunofenotipaje por citometría de flujo. Aquellos niños con carga viral inicial detectable para CMV fueron evaluados un año después (valoración cualitativa y oftalmológica). Correlación de Pearson, t Student. Se estudiaron 23 niños, 17 menores de 6 años; 21 de ellos (82.6 por ciento): lgG CMV +. Dos pacientes (8.7 por ciento): lgM CMV+ y promedio de carga viral: 11920 VID, dos IgM-IgG+, promedio de carga de 23129 VID; el resto negativos; todos con linfocitos CD4+ por encima del 25 por ciento. 50 por ciento de los niños con carga viral negativa para CMV, con contajes de CD4+ inferiores al 25 por ciento. El análisis de correlación de Pearson no mostró correlación entre la carga viral del VIH y los valores de VID para CMV (R²=0.13). Linfocitos CD8+: 32,3 ± 6,8 por ciento en los pacientes con carga para CMV, estadísticamente inferior al promedio del grupo sin cargas virales CMV: 49,1 ± 8,8. (t Student= 3,508; g.l: 17. P<0,003). Ningún niño presentó evidencia de enfermedad órgano específica (EOE), incluyendo retinitis. Independientemente de la presencia o no de IgM positiva para CMV, 4 pacientes tuvieron carga viral detectable. No hay correlación entre las cargas virales de ambos virus. Ningún niño con carga viral detectable para CMV...

In HIV-positive patients the diagnosis of CMV infection is essential. The relationship between serology, viral detection and clinical evolution has not been fully established. To detect the presence of CMV in peripheral blood by PCR testing in HIV positive pediatric patients with no infection symptoms, and to relate the results with serology and clinical evolution during a year follow up. Inclusion criteria: Children under twelve years of age, both genders, with a diagnosis of HIV infection, and receiving HAART therapy, written consent signed by parents. Anti CMV serology was performed by ELISA, semi-quantification of CMV in peripheral blood by PCR and immunophenotyping by flow-cytometry. Children with detectable initial viral load for CMV were submitted to aqualitative and ophtalmologic assessment one year later. Statistics: Pearson’s Correlation, Student’s t. 23 children, both genders, 17 under 6 years of age; 21 (82.6%): IgG CMV+. Two patients (8.7%): IgM CMV+ and viral load. 11920 IDV, two with IgM- IgG+, viral load average: 23129 IDV. The rest of the children were negative, all with lymphocytes CD4+ above 25%. 50% of the children had a negative viral load for CMV with CD4+ counts under 25%. There was no correlation between the HIV viral load and IDV values for CMV (r2=0.13). Lymphocytes CD8+32.3+ 6.8% in patients with viral load for CMV.This is statistically lower than the average for the group without viral loads CMV: 49.1 + 8.8 (Student’s t= 3.508; g.l: 17. p<0.003). No child showed evidence of specific organ disease (SOD), including retinitis. Regardless of serology for IgM, 4 patients had detectable viral loads. There was no correlation between the viral loads of the two viruses. No child with detectable viral load for CMV developed a specific organ disease, probably due to a highly efficient antiretroviral treatment. Viral load quantification for CMV in HIV + patients is recommended, regardless of specific IgM result.

A follow up study of cytomegalovirus infection in a group of Turkish renal transplant recipients using molecular assays

The clinical value of an in-house cytomegalovirus nested polymerase chain reaction (CMV-PCR) and a commercial molecular assay hybrid capture CMV DNA assay (HCA) was evaluated in monitoring a group of renal transplant patients for six months follow up. In this study, the sensitivity, specificity, positive predictive value, and negative predictive value of nested CMV DNA PCR assay and HCA at the beginning of the study were 70, 42.9, 46.7, 66.7, and 60, 78.6, 66.7, and 73.3 percent respectively. After six months, they were 80, 66.7, 80, 66.7 for CMV PCR and 73.3, 88.9, 91.7, 66.7 percent for HCA respectively. These results indicate that in monitoring and predicting CMV infections in renal transplant recipients, not only qualitative but also quantitative assays must be used together in order to decide the preemptive strategies.