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It is still not well known, in a population with high human cytomegalovirus (HCMV) seroprevalence, whether a child with congenital infection harbors multiple viral strains at birth, and whether the prolonged viral excretion in these children is secondary to the persistence of the same viral strain. To verify the genomic diversity of HCMV detected in congenitally infected children, the nucleotide viral sequences from urine and/or saliva obtained at birth from 14 newborns with congenital infection and breast milk obtained from mothers of 5 of these children were analyzed. Among the 14 children, 10 had sequential samples until the median age of 10 months. The viral nucleotide sequences in the breast milk were compared with those identified in the respective children at birth. The differentiation of viral strains was based on the variability of 3 regions of viral genes (UL55/gB, UL144, and UL73/gN). In 13/14 children (92.8%), a single genotype was observed at birth. Different viral genotypes were found in 1 child (7.2%). Among the sequential samples from 10 children, the same genotype obtained at birth was detected in 9/10 (90%), and in 1 of them (10%), a genotype change in the urine was found. More than 1 HCMV strain in milk was observed in 2 mothers (2/5, 40%). In a population with high seroprevalence, a single genotype was found in the majority of infected children. Reinfection did not frequently occur in the first months of life. Maternal reinfection does not seem to be a rare event in transmitter mothers.

Human cytomegalovirus (Human herpesvirus 5, HCMV) causes frequent asymptomatic infections in the general population. However, in immunosuppressed patients or congenitally infected infants, HCMV is related to high morbidity and mortality. In such cases, a rapid viral detection is crucial for monitoring the clinical outcome and the antiviral treatment. In this study, we optimized a sensitive biplex TaqMan® real-time PCR for the simultaneous detection and differentiation of a partial HCMV UL97 sequence and homologous extrinsic control (HEC) in the same tube. HEC was represented by a plasmid containing a modified HCMV sequence retaining the original primer binding sites, while the probe sequence was substituted by a phylogenetically divergent one (chloroplast CF0 subunit plant gene). It was estimated that the optimal HEC concentration, which did not influence the HCMV amplification is 1,000 copies/reaction. The optimized TaqMan® PCR demonstrated high analytical sensitivity (6.97 copies/reaction, CI = 95%) and specificity (100%). Moreover, the reaction showed adequate precision (repeatability, CV = 0.03; reproducibility, CV = 0.0027) and robustness (no carry-over or cross-contamination). The diagnostic sensitivity (100%) and specificity (97.8%) were adequate for the clinical application of the molecular platform. The optimized TaqMan® real-time PCR is suitable for HCMV detection and quantitation in predisposed patients and monitoring of the applied antiviral therapy. J. Med. Virol. 88:1604-1612, 2016. © 2016 Wiley Periodicals, Inc.

Objectives To estimate, by neonatal screening, the birth prevalence of congenital toxoplasmosis among live-born infants in Sergipe state, Brazil, and to investigate the clinical features of affected infants. Methods Dried blood spot specimens obtained from 15 204 neonates were assayed for the presence of anti-T. gondii IgM antibodies. Duplicate retesting was done in infants with positive and borderline results. Confirmatory testing in peripheral blood samples consisted of testing for anti-T. gondii IgG and IgM in infants and mothers. Those with possible congenital toxoplasmosis were evaluated and followed up to a median age of 20 months. Congenital infection was confirmed in the presence of persisting anti-T. gondii IgG antibodies beyond 12 months of age. All infants with confirmed infection were treated with pyrimethamine, sulfadiazine and folinic acid for 1 year. Results Fifty-three infants had detectable IgM in dried blood spot specimens. Confirmatory testing was reactive in 39/50, of which, 38 completed follow-up. Six of 15 204 newborns were diagnosed with congenital toxoplasmosis, resulting in an estimated birth prevalence of four per 10 000 [CI 95% 1.4–8.0]. Four infants (67%) showed signs of congenital toxoplasmosis in their first year of life; three (75%) had retinochoroidal scars, and one had cerebral calcifications. Two infants remained asymptomatic until 20 months of age. Conclusions The birth prevalence of congenital toxoplasmosis is high in the Brazilian state of Sergipe, with most of the infants showing ocular lesions. Preventive measures are strongly warranted. Objectifs: Estimer par le depistage neonatal, la prevalence a la naissance de la toxoplasmose congenitale chez les enfants nes vivants dans l’etat de Sergipe, au Bresil et etudier les caracteristiques cliniques des nourrissons atteints. Methodes: Des specimens seches de goutte de sang obtenus a partir de 15.204 nouveau-nes ont ete testes pour la presence d’anticorps IgM anti T. gondii. Un second test a ete effectue chez les nourrissons ayant des resultats positifs et intermediaires. Les tests de confirmation sur les echantillons de sang peripherique ont consistea tester pour les anticorps IgG et IgM anti T. gondii chez les nourrissons et les meres. Ceux atteints d’une eventuelle toxoplasmose congenitale ont eteevalues et suivis jusqu’a un âge median de 20 mois. L’infection congenitale a ete confirmee dans la presence de la persistance d’anticorps IgG anti T. gondii au dela de l’âge de 12 mois. Tous les nourrissons atteints d’une infection confirmee ont ete traites avec la pyrimethamine, la sulfadiazine et l’acide folinique pendant un an. Resultats: 53 enfants avaient des IgM detectables dans les echantillons seches de goutte de sang. Les tests de confirmation ont ete positifs dans 39/50 cas, dont 38 ont complete le suivi. Six des 15.204 nouveaux-nes ont ete diagnostiques avec une toxoplasmose congenitale, resultant en une prevalence a la naissance estimee a 4 pour 10.000 [IC95%: 1,4 a 8,0]. Quatre nourrissons (67%) presentaient des signes de la toxoplasmose congenitale au cours de leur premiere annee de vie, trois (75%) presentaient des cicatrices retinochoroidales et un presentait des calcifications cerebrales. Deux nourrissons sont restes asymptomatiques jusqu’a l’âge de 20 mois. Conclusions: La prevalence a la naissance de la toxoplasmose congenitale est elevee dans l’etat bresilien de Sergipe, avec la plupart des nourrissons presentant des lesions oculaires. Des mesures preventives sont fortement justifiees. Objetivos: Calcular, mediante un cribado neonatal, la prevalencia de nacimientos con toxoplasmosis congenita entre ninos nacidos vivos en el estado de Sergipe, en Brasil, e investigar las caracteristicas clinicas de los neonatos afectados. Metodos: Se obtuvieron muestras de sangre seca de 15,204 neonatos y se les realizaron pruebas para evaluar la presencia de anticuerpos IgM anti-T.gondii. Se realizo una prueba confirmatoria a los neonatos con resultados positivos y dudosos. Las pruebas confirmatorias en muestras de sangre periferica consistian en detectar las IgG e IgM especificas para T.gondii en neonatos y madres. Se evaluaron aquellos con una posible toxoplasmosis congenita y se les realizo un seguimiento hasta que tenian una edad media de 20 meses. Se confirmo una infeccion congenita en presencia anticuerpos IgG anti-T.gondii persistentes mas alla de los 12 meses de edad. Todos los neonatos con una infeccion confirmada eran tratados con pirimetamina, sulfadiazina y acido folinico durante un ano. Resultados: 53 neonatos tenian IgM detectables en las muestras de sangre seca. Las pruebas confirmatorias eran reactivas en 39/50 neonatos, de los cuales 38 cumplieron con el seguimiento. A seis de 15,204 neonatos se les diagnostico una toxoplasmosis congenita, resultando en una prevalencia estimada de nacimiento de 4 por 10,000 [IC 95% 1.4-8.0]. Cuatro neonatos (67%) presentaron signos de toxoplasmosis congenita durante su primer ano de vida; tres (75%) tenian cicatrices retinocoroideas, y uno tenia calcificaciones cerebrales. Dos neonatos se mantuvieron asintomaticos hasta los 20 meses de edad. Conclusiones: La prevalencia de nacimientos con toxoplasmosis congenita es alta en el estado Brasilero de Sergipe, con la mayoria de los neonatos presentando lesiones oculares. Las medidas preventivas estan ampliamente justificadas.

To evaluate cases of mother-to-child transmission of HIV-1 at multiple sites in Latin America and the Caribbean in terms of missed opportunities for prevention.Pregnant women infected with HIV-1 were eligible for inclusion if they were enrolled in either the NISDI Perinatal or LILAC protocols by October 20, 2009, and had delivered a live infant with known HIV-1 infection status after March 1, 2006.Of 711 eligible mothers, 10 delivered infants infected with HIV-1. The transmission rate was 1.4% (95% CI, 0.7-2.6). Timing of transmission was in utero or intrapartum (n=5), intrapartum (n=2), intrapartum or early postnatal (n=1), and unknown (n=2). Possible missed opportunities for prevention included poor control of maternal viral load during pregnancy; late initiation of antiretrovirals during pregnancy; lack of cesarean delivery before labor and before rupture of membranes; late diagnosis of HIV-1 infection; lack of intrapartum antiretrovirals; and incomplete avoidance of breastfeeding.Early knowledge of HIV-1 infection status (ideally before or in early pregnancy) would aid timely initiation of antiretroviral treatment and strategies designed to prevent mother-to-child transmission. Use of antiretrovirals must be appropriately monitored in terms of adherence and drug resistance. If feasible, breastfeeding should be completely avoided. Presented in part at the XIX International AIDS Conference (Washington, DC; July 22-27, 2012); abstract WEPE163.

Maternal antibodies may protect the fetus and neonate against severe forms of CMV-caused disease, therefore this study investigated the efficiency of the placental transfer of naturally acquired, maternal total anti-cytomegalovirus (CMV) IgG and neutralizing antibodies at different gestational ages. The study was conducted on 182 healthy CMV-seropositive Brazilian mothers and their 196 infants who were not infected congenitally with CMV, as determined by CMV detection in urine. The study groups were composed of 44 infants aged 28-30 weeks; 51 infants aged 31-33 weeks; 62 infants aged 34-36 weeks, and 39 infants of gestational age > or = 37 weeks. Quantitative detection of total CMV IgG was carried out using EIA and virus neutralizing titers were determined by a microneutralization assay in sera from mothers and infants. CMV IgG levels and neutralizing titers of the infants correlated with maternal levels (r=0.873 and r=0.841, respectively). The efficiency of placental transfer of these antibodies was enhanced significantly as gestation progressed until 34-36 weeks, when values similar to those of full-term infants (90-100%) were found. Transfer ratios were significantly higher for neutralizing compared to total CMV IgG antibodies at gestational age 31-33 weeks (100% vs. 84%, respectively) and at gestational age 28-30 weeks (75% vs. 60%, respectively). We conclude that placental transfer of naturally acquired maternal CMV neutralizing and total CMV IgG antibodies are similarly efficient above 34 weeks of gestational age. At less than 34 weeks of gestational age, transfer of neutralizing antibodies may be favored and these antibodies reach the neonatal serum of 99% of these premature infants.