Molecular analysis of rubella virus in travelers suspected of measles infection in São Paulo, Brazil / Análise molecular do vírus da rubéola em turistas suspeitos de infecção por sarampo em São Paulo, Brasil

OBJECTIVE: To identify measles virus genotypes in three cases of travelers suspected of measles infection. METHODS: Samples (blood and urine) were collected for serology, virus isolation, and genotyping. Sera were analyzed for IgM antibodies against measles virus and rubella virus by enzyme-linked immunosorbent assay (ELISA) (Siemens - Marburg, Germany). Clinical samples (lymphocytes and urine) were inoculated into Statens Serum Institute rabbit corneal epithelial cell line- ATCC CL 60 (SIRC) and Vero Slam cells. RNA was extracted from clinical samples and cell culture was inoculated and processed by polymerase chain reaction (PCR) with oligonucleotides specific for measles virus (MV) and rubella virus (RV). RESULTS: All patients showed IgM negative serology for MV and positive IgM for RV. RV belonging to genotypes 1B, 1C, and 1E were isolated from patients who came from Finland, Peru, and Germany, respectively. Genotype 1B has been found in Europe and on the East Coast of South America; 1C has been found in Peru and the West Coast of South America, and 1E, first identified in 1997, now appears to have worldwide distribution. CONCLUSION: Information about RV and MV genotypes circulating in São Paulo is essential for the control of measles, rubella, and congenital rubella syndrome (CRS) in Brazil.

OBJETIVO: Identificar o genótipo do vírus do sarampo em três viajantes suspeitos de infecção por sarampo. MÉTODOS: Amostras (sangue e urina) foram coletadas para sorologia, isolamento viral e genotipagem. As sorologias para pesquisa de IgM para o vírus do sarampo e da rubéola foi realizada utilizando-se o kit de ELISA (Siemens - Marburg, Alemanha). As amostras clínicas (linfócito e urina) foram inoculadas na SIRC (Statens Serum Institute rabbit corneal epithelial cell line-ATCC CL 60) e nas células Vero Slam. O RNA foi extraído das amostras clínicas e das células inoculadas e processadas por PCR, utilizando oligonucleotideos específicos para sarampo e rubéola. RESULTADOS: Todos os pacientes apresentaram sorologia IgM negativa para sarampo e positivo para rubéola. Os vírus da rubéola isolados dos pacientes que vieram da Finlândia, Peru e Alemanha pertencem aos genótipos 1B, 1C e 1E, respectivamente. O genótipo 1B foi encontrado na Europa e na costa oriental da América do Sul, o genótipo 1C foi encontrado no Peru e na costa oeste da América do Sul e o genótipo 1E, identificado pela primeira vez em 1997, agora aparenta ser um genótipo com distribuição mundial. CONCLUSÃO: O conhecimento dos genótipos de sarampo e rubéola que circulam em São Paulo é essencial para o controle do sarampo, rubéola e síndrome da rubéola congênita.

Relative efficiency of polymerase chain reaction and enzyme-linked immunosorbant assay in determination of viral etiology in congenital cataract in infants.

BACKGROUND: Perinatal viral infections of fetus are among the leading causes of congenital cataract and identifying the viral etiology is important. OBJECTIVES: To detect the presence of Rubella virus (RV), herpes simplex virus (HSV) and cytomegalovirus (CMV) in lens aspirate specimens obtained from patients with congenital cataract and relate the results with serology. SETTING AND DESIGN: Prospective study carried out in tertiary care hospital. MATERIALS AND METHODS: Fifty lens aspirates from 50 infants with congenital cataract were subjected to HSV, RV isolation and polymerase chain reaction (PCR) for detection of HSV and CMV. Reverse transcription polymerase chain reaction (RT-PCR) was applied for RV detection. Peripheral blood specimens were screened for anti-HSV, RV and CMV antibodies by enzyme-linked immunosorbant assay (ELISA). RESULTS: Rubella virus was detected in nine (18%) lens aspirates, by nRT-PCR which includes six positive by culture. HSV-2 DNA was detected in nine other lens aspirates, while CMV was not detected by PCR. Serological results did not correlate with the presence of viruses in the lens aspirates. This is the first report of detection of HSV-2 DNA in cases of congenital cataract. CONCLUSIONS: Cytomegalovirus may not be playing a significant role in causation of congenital cataract. The role of serology in identifying causative viral infection for congenital cataract needs to be re-evaluated.

Nested reverse transcription polymerase chain reaction for the detection of rubella virus in clinical specimens.

BACKGROUND & OBJECTIVE: Rubella virus (RV) is one of the leading causes of childhood blindness in India. In this study we applied an optimized nested reverse transcription polymerase chain reaction (nRT-PCR) to detect RV in clinical specimens. METHODS: nRT-PCR was optimized using total RNA extracted from standard strain of RV using nested sets of primers specific for E1 open reading frame. nRT-PCR was applied onto 30 lens aspirates and corresponding peripheral blood leucocytes of 30 infants with congenital (29)/ developmental (01) cataract. Serology for anti-RV IgG and IgM antibodies was done. RV isolation was attempted using Vero and SIRC cell cultures. RESULTS: Optimized nRT-PCR was specific for RV and sensitive to detect 10 fg of RV RNA. Among 30 patients, nRT-PCR detected presence of RV in lens aspirates of 6 (20%) and 4 corresponding leucocytes. RV was isolated from 3 (10%) lens aspirates (nRT-PCR positive) of the 30 patients. Sera of these 6 patients showed presence of anti-RV IgG and IgM in one, only anti-RV IgG in 3 others and none in the other two. Of the remaining 24 patients, anti-RV IgG was detected in 3 and no anti-RV IgM antibodies in others. INTERPRETATION & CONCLUSION: Findings of our study showed that the nRT-PCR was a more sensitive and rapid technique to detect RV from lens aspirates compared to conventional methods of virus isolation and serology.

Campaña nacional de vacunación para la eliminación de la rubéola y del síndrome de rubéola congénita en Bolivia, 2006 / National vaccination campaign for the elimination of rubella and congenital rubella syndrome in Bolivia, 2006

En mayo 2006, Bolivia llevó a cabo una campaña nacional de vacunación en su población de 15 a 39 años para eliminar la rubéola y el síndrome de rubéola congénita del país. Durante seis semanas el país sumó los esfuerzos de los políticos, aliados nacionales e internacionales, trabajadores de salud y organizaciones sociales para alcanzar la meta. La cobertura nacional validada por una encuesta post campaña fue de 94%, sin diferencias significativasde edad, sexo o en la distribución rural urbana. La encuesta probó también un alto nivel de confianza de la población en los servicios de vacunación y el rol importante del personal de salud para informar a la población en el medio rural. Se demuestra que actividades de vacunación masivas en la población adulta y de ambos sexos pueden ser exitosas con una preparación minuciosa y un plan de comunicación estudiado.

Estudio de un mecanismo de persistencia del virus Rubéola en la infección congénita / Study of a mechanism of persistence of the virus Rubella in the congenital infection

Se investigó el rol de la apoptosis en un posible mecanismo de persistencia del virus Rubéola (Rub) en la infección congénita, utilizando cultivos primarios de fibroblastos fetales humanos (FFH) en activa proliferación. Se demostró que, en contraste con lo que se observa en células Vero, RK13, placenta humana a término, y fibroblastos diploides de adulto humano (Hs888Lu), Rub no induce apoptosis en FFH. Por inmunohistoquímica se detectó la actividad de JUN, gen que promueve la proliferación celular, en FFH pero no en Hs888Lu. El análisis de expresión de genes en FFH y Hs888Lu con microarreglos de ADN mostró que en FFH están sobre-expresados genes que estimulan la supervivencia y proliferación celular, como factores de crecimiento, PI3K, y oncogenes de la familia Ras. Las kinasas p-Akt y p-ERK se detectaron en FFH por western blotting. A continuación se demostró que Rub sí induce apoptosis en FFH, cuando se suprimen las vías PI3K–Akt y Ras–Raf–MEK–ERK. Significativamente, la infección de las células fetales no prospera en estas condiciones. Así, la persistencia de Rub en la infección congénita puede estar inversamente relacionada con la ocurrencia de apoptosis en las células hospedadoras. También se analizó la expresión de genes en células infectadas por Rub, identificándose más de 1500 genes celulares inducidos o suprimidos por el virus, muchos de ellos relacionados con la respuesta inmune, las vías metabólicas, y las cascadas de transmisión de señales. En FFH, Rub modifica la expresión de diferentes genes del desarrollo (genes Hox), que establecen el patrón corporal y promueven la proliferación y la diferenciación celular durante la morfogénesis. Estos datos representan nuevos puntos de partida para postular hipótesis sobre el modo en que el virus ejerce su efecto teratogénico, y brindan un amplio panorama para estudiar la relación entre Rub y la célula.