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.

Neonatal cytomegalovirus infection: diagnostic modalities available for early disease detection.

CMV is a ubiquitous virus. In India, there is high seroendemicity with almost 99% adults showing IgG antibodies. Infection or re-activation becomes important in immunocompromised host (Transplant recipients, Cancer therapy patients and patients with HIV/AIDS). Neonates form a distinctive high risk population for congenital CMV infection and suffer disastrous sequlae of the same. Neonatal infections may be congenital in nature or may be acquired after birth during first month of life via infected breast milk or due to exposure to high risk blood products. The risk for transmission of the virus to the fetus is higher in primary infected mothers than in mothers with reactivated disease. Primary CMV infections are reported in 1-4% of seronegative women during pregnancy and the risk for viral transmission to fetus is 30-40%. Reactivation of a CMV infection during pregnancy is reported in 10-30% of seropositive women and the risk of transmitting the virus is about 1-3%. The adverse outcome of congenital neonatal CMV infection includes- microcephaly (70%), intellectual impairment (60%), sensineural hearing loss (35%), choriorenitis (22%), hepatosplenomegaly (70%), jaundice (68%), thrombocytopenia (65%), low birth weight (65%), pneumonitis (2-5%) and congenital heart disease (<5%). About 5-10% of congenitally infected asymptomatic infants will have neurological problems later in life the most common of which is unilateral or bilateral sensory neural hearing loss. All immunocompromised hosts, including pre-term neonates, mount weak antibody responses (IgM), making serological detection of CMV infection in them, fallacious. Thus, it is imperative to use antigen detection methods such as quantitative PCR or PP65 Antigenaemia assays to detect CMV infection in immunocompromised host. Sakhuja et al and Minz et al have demonstrated that PP65 Antigenaemia assay is very good for diagnosing CMV disease in renal transplant recipients. The present review tends to highlight the role of newer diagnostic modalities in early CMV infection detection in neonatal population.

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 serological study of cytomegalovirus infection in pregnant and non-pregnant women at Gauhati Medical College and Hospital.

Primary cytomegalovirus infection is the most common infection during pregnancy that may have long-term neurodevelopmental sequelae in children born to these mothers. It is also associated with many obstetric complications. So the aim of this study was to determine the seroprevalence of cytomegalovirus infection in local antenatal population with bad obstetric history and to see the effects, if any, of age, socio-economic status, presenting features and different gestational periods. Seventy-five pregnant women with bad obstetric history were screened for the presence of cytomegalovirus specific IgM and IgG antibodies by doing enzyme-linked immunosorbent assay, out of which 17(22.66%) had evidence of recurrent cytomegalovirus infection as demonstrated by the presence of cytomegalovirus specific IgM antibodies. All were found to be positive for cytomegalovirus specific IgG antibodies. This indicates that the presence of cytomegalovirus specific IgM antibodies in this population is an evidence of reactivation of a latent infection or re-infection with a different strain of cytomegalovirus. Increased IgM seropositivity was found to be associated with advancing age, poor, socio-economic status, third trimester of pregnancy and bad obstetric history like premature delivery, stillbirth, recurrent spontaneous abortions, intra-uterine growth retardation. Out of 25 randomly selected non-pregnant women of childbearing age, all showed presence of cytomegalovirus specific IgG antibodies and none was found to be positive for primary or recurrent cytomegalovirus infection.

Transfusion-related Cytomegalovirus Infection Among Very Low Birth Weight Infants in an Endemic Area

This study investigated the incidence of acquired cytomegalovirus (CMV) infection in very low birth weight infants (VLBWI) given CMV seropositive blood, and sought to determine whether filtering and irradiation of blood products could help prevent CMV infection and the time required to clear passively-derived anti-CMV IgG among 80 VLBWI transfused with filtered-irradiated blood, 20 VLBWI transfused with nonfiltered- nonirradiated blood and 26 nontransfused VLBWI. CMV IgG and IgM values were obtained from all blood products prior to transfusions, and from VLBWI at birth until the infants became seronegative. Urine was obtained for CMV culture at birth and every 3-4 weeks until 12 weeks after the final transfusion. The incidence of CMV IgG seropositivity among the 126 infants at birth and the blood products given were 96% and 95%, respectively. The incidence of acquired CMV infection was 4/100 (4%) in the transfused group: 2/80 (2.5%) and 2/20 (10%) in the filtered-irradiated and nonfiltered-nonirradiated transfusion groups, respectively. Approximately 9-10 months elapsed to clear passively acquired CMV IgG. The irradiation and filtering of the blood products did not seem to decrease the transfusion-related CMV infection rate in Korea among VLBWI, however, further validation is recommended in a larger cohort of infants.

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.

Diagnóstico etiológico de pacientes con exantemas o cuadros febriles atendidos en 1998. Estado Zulia, Venezuela / Laboratory diagnosis of patients with exanthematic or febrile syndromes occurring in the Zulia State, Venezuela, during 1998

BACKGROUND: Exanthematic diseases are a group of syndromes mainly caused by acute viral infections. AIM: To obtain information about the viruses that cause exanthematic diseases in our region. PATIENTS AND METHODS: During 1998, 267 serum samples from patients with an acute rash or patients presenting a febrile syndrome accompanied by enlarged lymph nodes, headache and other symptoms, were collected. Specific antibody of the IgM class (anti-IgM) against Rubella, Measles, Dengue types 1-4 and Cytomegalovirus (CMV) were measured by immunoenzymatic assay (EIA). Epstein-Barr virus (EBV) antibodies were measured by immunofluorescence. RESULTS: An etiologic agent was detected in 208 cases (77.9%). Cases due to Dengue (40.6%) and Rubella (21%) viruses predominated, but the frequency of other agents was also high in specific age groups. The agreement between the clinical suspicion and the laboratory findings varied broadly, from a 100% for suspected Dengue to just a 14.8% for cases of suspected CMV infection. CONCLUSIONS: Dengue was the most common viral exanthematic disease in the Zulia State during 1998.

Comparing serology, antigenemia assay and polymerase chain reaction for the diagnosis of cytomegalovirus infection in renal transplant patients.

BACKGROUND: Cytomegalovirus (CMV) disease is responsible for significant morbidity and mortality following renal transplantation. Currently serology is the only method widely available in our country. Newer methods like early CMV pp65 antigenemia assay and CMV DNA amplification can diagnose CMV disease in its very early period. AIM: The aim of our study was to compare serologic method with antigenemia assay and CMV DNA amplification to diagnose CMV. METHODS: Seventy-three renal transplant recipients (from 7 centres) with clinical suspicion of CMV disease were studied prospectively. The diagnosis of CMV infection was suspected on the basis of fever and leucopenia. RESULT AND DISCUSSION: Three tests were done in all 73 patients and in 22 healthy subjects (control group). The sensitivity and specificity of serological test (CMV IgM) was 72.97 and 62.06%; of antigenemia assay was 89.18 and 100% and of PCR was 100 and 72.41%. CONCLUSION: Antigenemia assay is a sensitive and specific test for early and rapid diagnosis of CMV infection. Qualitative PCR is a sensitive marker but has low specificity.

Cytomegalovirus infection in renal transplant recipients diagnosed by nested-PCR

A prospective study of cytomegalovirus (CMV) infection was carried out on 34 renal transplant recipients managed at a General Hospital in Ribeirão Preto, SP, Brazil. Serologic tests showed that all patients were infected with CMV before renal transplantation. Two nested-PCR techniques with primers that recognize sequences of the glycoprotein B (gB) and H (gH) genes were used for CMV detection in blood and urine samples during the post-transplantation period. CMV was detected more frequently in blood samples than in urine samples (P<0.001). Thirty-three patients had CMV detected at least once in blood and/or urine samples. Seven of these patients (21.2 percent) were diagnosed as having symptomatic CMV infection and showed a worse clinical outcome, with a higher death rate (P = 0.03). No association between CMV viremia and graft rejection was observed. Nested-PCR was not useful to identify patients at risk for symptomatic CMV infection since only 21.2 percent of the patients with CMV infection were symptomatic

Seroprevalence of cytomegalovirus infection in children.

A hospital-based cross-sectional survey was conducted in Bhumibol Adulyadej Hospital between February and May 1997 to study the seroprevalence of cytomegalovirus (CMV) infection in hospitalized infants. Of 83 cases, 46 were boys and 37 girls, with the mean age of 11.87 months. The seroprevalence of CMV infection was 80.95, 61.90, 80.95 and 70.00% in infants at the age range of 0-6, 6-12, 12-18 and 18-24 months respectively. After excluding infants below 6 months of age, the seroprevalence rate was 70.97%. The family income of infants with positive CMV antibody was significantly lower than that of infants with negative results. There were no statistical correlations between seroconversion and age, sex, number of children in family and place of child rearing.

Gene cloning of the human cytomegalovirus (HCMV) antigen reactive with the serum from a HCMV-infected patient

The human cytomegalovirus(HCMV) gene encoding the protein reactive with the sera of HCMV-infected patient was cloned and characterized. A reactive phage clone was screened from a lambda gt11 expression library of cDNA of HCMV AD169 strain using HCMV-infected patient sera. The recombinant protein was expressed as 138 kDa-fusion protein with beta-galactosidase, which was reactive with IgM or IgG HCMV antibody-positive sera, but not with anti-HCMV antibody-negative sera. A homology search of the DNA sequence of the cloned gene with HCMV AD169 sequences revealed that it was composed of 709 base pairs spanning between 0.174 and 0.177 map units of the UL32 region of the HCMV AD169 strain genome. This position corresponded to a part of the gene encoding 150 kDa phosphoprotein-(pp150), a major tegument protein, which was reported as an immunogenic protein which evoked strong and longstanding antibody response and had no sequence homology with the proteins of other herpesviruses. These results suggested that pp150 was an immunogenic protein in natural HCMV infection and therefore this clone was regarded as a useful candidate for developing an antigen for the serodiagnosis of HCMV.

Infecção pelo citomegalovirus em pacientes com sindrome da imunodeficiencia adquirida (AIDS): relacoes clinico-virologicas e anatomopatologicas / Infection by cytomegalovirus in patients with acquired immunodeficiency syndrome (AIDS): clinical, virological and histopathological correlations

Com o objetivo de determinar a prevalencia da infeccao pelo Citomegalovirus (CMV) em pacientes com AIDS, bem como relacionar os achados clinicos virologicos decorrentes desta infeccao com as repercussoes anatomopatologicas, estudamos 50 pacientes adultos atendidos entre abril de 1986 a junho de 1987, em dois hospitais publicos de Sao Paulo (HSP e HSPE). Estes pacientes foram acompanhados clinica e laboratorialmente, por periodo medio de 2 meses com coletas seriadas de sangue, urina e saliva. Foram realizados isolamento do CMV em monocamadas de fibroblastos humanos e testes sorologicos de Imunofluorescencia Indireta (IFI-IgG/IgM) e Reacao Imunoenzimatica (ELISA-IgG). No momento da admissao no estudo 20 por cento (10/50) dos pacientes apresentavam anticorpos IgM CMV especificos e 100 por cento (50/50) deles anticorpos IgG (IFI). Durante o acompanhamento, 5 pacientes inicialmente IgM negativos tornaram-se IgM positivos, sugerindo reativacao ou reinfeccao pelo CMV. O CMV foi isolado de sangue periferico em 12,5 por cento, da urina em 23,2 por cento, da saliva em 21,9 por cento dos pacientes. Exames anatomopatologicos foram realizados em 24 pacientes, correspondendo a 60 por cento dos pacientes que evoluiram para obito durante o periodo de estudo...