Molecular evolution of coxsackievirus A24v in Cuba over 23-years, 1986-2009.

Coxsackievirus A24 variant (CVA24v) is a major causative agent of acute hemorrhagic conjunctivitis outbreaks worldwide, yet the evolutionary and transmission dynamics of the virus remain unclear. To address this, we analyzed and compared the 3C and partial VP1 gene regions of CVA24v isolates obtained from five outbreaks in Cuba between 1986 and 2009 and strains isolated worldwide. Here we show that Cuban strains were homologous to those isolated in Africa, the Americas and Asia during the same time period. Two genotypes of CVA24v (GIII and GIV) were repeatedly introduced into Cuba and they arose about two years before the epidemic was detected. The two genotypes co-evolved with a population size that is stable over time. However, nucleotide substitution rates peaked during pandemics with 4.39 × 10-3 and 5.80 × 10-3 substitutions per site per year for the 3C and VP1 region, respectively. The phylogeographic analysis identified 25 and 19 viral transmission routes based on 3C and VP1 regions, respectively. Pandemic viruses usually originated in Asia, and both China and Brazil were the major hub for the global dispersal of the virus. Together, these data provide novel insight into the epidemiological dynamics of this virus and possibly other pandemic viruses.

Vigilancia de laboratorio de dengue y otros arbovirus en Cuba, 1970-2017 / Laboratory surveillance of dengue fever and other arboviruses in Cuba, 1970-2017

Los arbovirus constituyen una de las principales causas de emergencia en salud por la morbilidad y mortalidad que producen y el estrés sanitario que conllevan. Cuba no ha estado excenta de riesgo, y el enfrentamiento del dengue inicialmente y de otros arbovirus después, ha sido, y es, una prioridad de las máximas autoridades del país. La vigilancia de laboratorio de dengue se estableció desde inicios de la década del 70 aunque sus objetivos y estrategias han cambiado según la situación epidemiológica nacional y regional y la tecnología de diagnóstico disponible. Se destacan cuatro etapas en su desarrollo. En este trabajo se resumen las estrategias desarrolladas para la vigilancia de laboratorio de dengue y de otros arbovirus en el periodo de 1970 a 2017. Se describe además el papel desempeñado por el Instituto de Medicina Tropical, ¨Pedro Kouri¨ (IPK) como Laboratorio Nacional de Referencia(AU)

Arboviruses are one of the leading causes of health emergencies due to their morbidity and mortality and the sanitary stress they bring about. Cuba has not been free from risk, and the response first to dengue fever and then to other arboviruses has been and still is a priority for the country's top authorities. Laboratory surveillance of dengue fever was implemented in the 1970s, though its aims and strategies have evolved in keeping with the national and regional epidemiological situation, and the available diagnostic technology. Four stages stand out in the development of dengue laboratory surveillance. The present paper summarizes the strategies developed for laboratory surveillance of dengue fever and other arboviruses in the period 1970-2017. A description is also provided of the role played by Pedro Kourí Tropical Medicine Institute (IPK) as a national reference laboratory(AU)

Coxsackievirus A6 and enterovirus 71 causing hand, foot and mouth disease in Cuba, 2011-2013.

Hand, foot and mouth disease (HFMD) is usually caused by coxsackievirus A16 or enterovirus 71 (EV71). Between 2011 and 2013, HFMD cases were reported from different Cuban provinces. A total of 42 clinical specimens were obtained from 23 patients. Detection, identification and phylogenetic analysis of enterovirus-associated HFMD were carried out by virus isolation, specific enterovirus PCR and partial VP1 sequences. HEV was detected in 11 HFMD cases. Emerging genetic variants of coxsackievirus A6 and EV71 were identified as the causative agents of the Cuban HFMD cases.

Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation.

Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36%) were positive at 24 h, 10 (40%) were positive at 48 h and six (24%) were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation

Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36%) were positive at 24 h, 10 (40%) were positive at 48 h and six (24%) were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

Normalización de un sistema de reacción en cadena de la polimerasa en tiempo real para la cuantificación de papilomavirus humano de alto riesgo oncogénico / Standardization of a real-time based polymerase chain reaction system for the quantification of human papilomavirus of high oncogenic risk

El objetivo de este trabajo fue normalizar e implementar un sistema de reacción en cadena de la polimerasa en tiempo real, para determinar la carga viral de 7 genotipos de papilomavirus humano (PVH) de alto riesgo oncogénico. Se evaluó la especificidad del sistema y se construyeron las curvas estándar para PVH 16 y 18, que se emplearon para la cuantificación de ADN viral en diferentes muestras de pacientes identificados como positivos a PVH, mediante reacción en cadena de la polimerasa (RCP) cualitativa y secuenciación nucleotídica. Se obtuvieron dos curvas estándar para PVH 16 y 18, a partir del ADN genómico de las líneas celulares SiHa y HeLa, las que mostraron una buena correlación lineal ( r = -0,99) y valores bajos de error. El límite inferior de detección a partir del ADN de las líneas celulares fue de hasta 10 copias para ambos genotipos. No se obtuvo reacción cruzada entre los diferentes tipos de PVH ni con otros virus ADN. La reacción en cadena de la polimerasa en tiempo real (RCP-TR) normalizada probó ser un sistema simple, rápido, específico y altamente sensible. Además, permitirá desarrollar investigaciones sobre la prevalencia de infección por PVH en Cuba, con vistas a la aplicación de las vacunas que se encuentran disponibles en el mercado internacional, así como la evaluación de otros candidatos vacunales diseñados en el futuro(AU)

The objective of the present study is to standardize a real-time based polymerase chain reaction system in order to detect and quantify 7 high risk human papillomavirus in different clinical samples from patients suspected of this type of infection. The validation of a 5´ exonuclease fluorescent probe real-time PCR assay (TaqMan format) for the detection and quantification of the 7 most frequent HR-HPV types (16, 18, 31, 33, 45, and 58) which account for over 87% of cervical carcinomas world-wide was carried out. Simultaneous PCR reactions are required to detect the designated HPV types. Specificity tests for each HPV type and other DNA viruses were performed. Standard external curve constructions were achieved, which allow determining the number of target DNA copies in the previously HPV tested samples. HPV 16 and 18 standard curves were obtained from purified genomic DNA of SiHa and HeLa cell lines, respectively. The pattern curves were constructed on the basis of each of the resulting standard DNA, which showed good linear correlation (r = -0, 99) and low error values. The lower detection limit was 10 copies for both HPV 16 and 18. No cross reactions between HPV types and other DNA viruses were observed. Real-Time Polymerase Chain Reaction system, standardized for 7 HPV types, proved to be a rapid, specific and highly sensitive system for better diagnosis and follow-up of patients with high grade intraepithelial lesions. In addition, this assay will allow the development of coming researches in relation with the prevalence and pathogenesis of human papillomavirus infections in different samples from Cuban patients(AU)

Epidemiological studies on dengue virus type 3 in Playa municipality, Havana, Cuba, 2001-2002.

OBJECTIVES: Recognizing the uniqueness of secondary dengue virus (DENV)-1/3 dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) cases at an interval of 24 years, we sought to estimate DENV infections as well as the ratios between mild disease and DHF/DSS by DENV infection sequence in Playa District (Havana, Cuba) during the 2001-2002 outbreak of dengue virus type 3 (DENV-3). METHODS: A retrospective seroepidemiological study was conducted in 2003 in Playa District. Blood samples were collected from a 1% random sample of residents and were studied for the prevalence of dengue neutralizing antibodies. RESULTS: DENV-3 was found to have infected 7.2% (95% confidence interval (95% CI) 6.0-8.4%) of susceptible individuals (the entire cohort), the majority of whom experienced silent infections. Virtually every individual who had a secondary infection in the sequence DENV-1 then DENV-3 became ill, with a ratio of severe to mild cases of 1:35 (95% CI 1:67-1:23). Secondary infections in the sequence DENV-2/3 were less pathogenic than DENV-1/3. Mild disease accompanying secondary DENV2/3 occurred at a ratio of 1:4.49 infections (95% CI 1:5.77-1:3.42) secondary infections. CONCLUSIONS: The results obtained highlight the role of the infecting serotype and also the sequence of the viral infection in the clinical outcome of a dengue infection.

Casos de poliomielitis paralítica asociada a la vacuna oral antipoliomielítica en Cuba (1963-2006) / Cases of paralytic poliomyelitis associated with oral antipoliomielytis vaccine in Cuba (1963-2006)

Objetivo: divulgar los indicadores de poliomielitis paralítica asociada encontrados en 44 años en niños vacunados con oral antipoliomielítica, suministrada exclusivamente en campañas masivas de vacunación durante el período de 1963 a 2006 y reportar el riesgo de poliomielitis paralítica asociada a la vacuna con relación a diferentes aspectos epidemiológicos. Métodos: se realizó un estudio retrospectivo en 596 casos de parálisis fláccida aguda en niños ingresados en hospitales pediátricos, basado en investigaciones virológicas y detección de diferentes variables epidemiológicas. Resultados: de 113 pacientes estudiados se aislaron 120 agentes virales y 30 fueron identificados como poliovirus. Los datos clínicos y epidemiológicos en 596 casos de parálisis fláccida aguda permitieron categorizar a 20 niños afectados con poliomielitis paralítica asociada a la vacuna. Todos los casos se presentaron en niños menores de un año, vacunados con oral antipoliomielítica con estrategias de campañas masivas de vacunación exclusivamente y 19 fueron producidos por la primera dosis. El riesgo global en los niños vacunados con primera dosis de 1963 a 2006 fue 1 en 379 888 (7 217 866 dosis administradas/19 casos con poliomielitis paralítica asociada a la vacuna). Los casos de poliomielitis paralítica asociada a la vacuna se han presentado esporádicos o en un grupo de 8 casos en el período 1989-1992. El riesgo de primera dosis en casos esporádicos fue 1 en 612 864 y en el grupo de 1 en 84 670. El riesgo en casos agrupados es 7,2 veces mayor que los ocurridos en casos aislados. Particularmente en el año 1992, que coincidió con una epidemia de neuropatía epidémica, el riesgo fue de 1 en 52 140, lo que representó un incremento de 11,8 veces a lo ocurrido en casos esporádicos. Los niños de 4-7 meses de edad también tuvieron un riesgo mayor que fue 1 en 132 812. Conclusiones: se identificaron aspectos epidemiológicos que incrementaron el riesgo de poliomielitis paralítica asociada a la vacuna, en los cuales coinciden aspectos de deficiencias nutricionales.

Aim: to disseminate the indicators of associated paralytic poliomyelitis found during 44 years in children that received the oral antipoliomyelitis vaccine, which was only administered in massive vaccination campaigns from 1963 to 2006, and to report the risk of vaccine-associated paralytic poliomyelitis as regards different epidemiological aspects. Methods: a retrospective study was undertaken in 596 cases of acute flaccid paralysis in children admitted in pediatric hospitals, based on virology researches, and on the detection of different epidemiological variables. Results: 120 viral agents were isolated from 113 studied patients. 30 were identified as poliovirus. The clinical and epidemiological data from 596 cases of acute flaccid paralysis allowed to categorize 20 children affected with vaccine-associated paralytic poliomyelitis. All the cases were children under one that were exclusively administered the oral antipoliomyelitis vaccine through the strategy of the massive vaccination campaigns. 19 of them were caused by the first dose. Global risk in children vaccinated with the first dose from 1963-2006 was of one in 379 888 (7 217 866 doses administered/19 cases with vaccine-associated paralytic poliomyelitis). Cases of vaccine-associated paralytic poliomyelitis have been sporadic or in a group of 8 cases during 1989-1992. The risk of first dose in sporadic cases was of one in 612 864, and in the group of 1 in 84 670. The risk of grouped cases is 7.2 times higher than those occurred in isolated cases. Particularly, in 1992, coinciding with an outbreak of epidemic neuropathy, the risk was of one in 52 140, which represented an increase of 11.8 times compared with sporadic cases. Children aged 4-7 months old also had a higher risk of 1 in 132 812. Conclusions: there were identified epidemiological aspects that augmented the risk of vaccine-associated paralytic poliomyelitis, in which the aspects of nutritional deficiencies coincided.

Casos de poliomielitis paralítica asociada a la vacuna oral antipoliomielítica en Cuba (1963-2006) / Cases of paralytic poliomyelitis associated with oral antipoliomielytis vaccine in Cuba (1963-2006)

Objetivo: divulgar los indicadores de poliomielitis paralítica asociada encontrados en 44 años en niños vacunados con oral antipoliomielítica, suministrada exclusivamente en campañas masivas de vacunación durante el período de 1963 a 2006 y reportar el riesgo de poliomielitis paralítica asociada a la vacuna con relación a diferentes aspectos epidemiológicos. Métodos: se realizó un estudio retrospectivo en 596 casos de parálisis fláccida aguda en niños ingresados en hospitales pediátricos, basado en investigaciones virológicas y detección de diferentes variables epidemiológicas. Resultados: de 113 pacientes estudiados se aislaron 120 agentes virales y 30 fueron identificados como poliovirus. Los datos clínicos y epidemiológicos en 596 casos de parálisis fláccida aguda permitieron categorizar a 20 niños afectados con poliomielitis paralítica asociada a la vacuna. Todos los casos se presentaron en niños menores de un año, vacunados con oral antipoliomielítica con estrategias de campañas masivas de vacunación exclusivamente y 19 fueron producidos por la primera dosis. El riesgo global en los niños vacunados con primera dosis de 1963 a 2006 fue 1 en 379 888 (7 217 866 dosis administradas/19 casos con poliomielitis paralítica asociada a la vacuna). Los casos de poliomielitis paralítica asociada a la vacuna se han presentado esporádicos o en un grupo de 8 casos en el período 1989-1992. El riesgo de primera dosis en casos esporádicos fue 1 en 612 864 y en el grupo de 1 en 84 670. El riesgo en casos agrupados es 7,2 veces mayor que los ocurridos en casos aislados. Particularmente en el año 1992, que coincidió con una epidemia de neuropatía epidémica, el riesgo fue de 1 en 52 140, lo que representó un incremento de 11,8 veces a lo ocurrido en casos esporádicos. Los niños de 4-7 meses de edad también tuvieron un riesgo mayor que fue 1 en 132 812....(AU)

Aim: to disseminate the indicators of associated paralytic poliomyelitis found during 44 years in children that received the oral antipoliomyelitis vaccine, which was only administered in massive vaccination campaigns from 1963 to 2006, and to report the risk of vaccine-associated paralytic poliomyelitis as regards different epidemiological aspects. Methods: a retrospective study was undertaken in 596 cases of acute flaccid paralysis in children admitted in pediatric hospitals, based on virology researches, and on the detection of different epidemiological variables. Results: 120 viral agents were isolated from 113 studied patients. 30 were identified as poliovirus. The clinical and epidemiological data from 596 cases of acute flaccid paralysis allowed to categorize 20 children affected with vaccine-associated paralytic poliomyelitis. All the cases were children under one that were exclusively administered the oral antipoliomyelitis vaccine through the strategy of the massive vaccination campaigns. 19 of them were caused by the first dose. Global risk in children vaccinated with the first dose from 1963-2006 was of one in 379 888 (7 217 866 doses administered/19 cases with vaccine-associated paralytic poliomyelitis). Cases of vaccine-associated paralytic poliomyelitis have been sporadic or in a group of 8 cases during 1989-1992. The risk of first dose in sporadic cases was of one in 612 864, and in the group of 1 in 84 670. The risk of grouped cases is 7.2 times higher than those occurred in isolated cases. Particularly, in 1992, coinciding with an outbreak of epidemic neuropathy, the risk was of one in 52 140, which represented an increase of 11.8 times compared with sporadic cases. Children aged 4-7 months old also had a higher risk of 1 in 132 812. Conclusions: there were identified epidemiological aspects that augmented the risk of vaccine-associated paralytic poliomyelitis, in which the aspects of nutritional deficiencies coincided(AU)

Neutralizing antibody response variation against dengue 3 strains.

To evaluate the neutralizing antibody activity of a human sera panel against seven strains of the homotypic virus. Sera were collected from DENV-3 immune individuals. Two DENV-3 genotypes and strains isolated at different time-points during the 2000 and 2001-2002 Havana epidemics were included. A panel of 20 late convalescent sera collected 16-18 months after acute illness from DF and DHF patients are studied. These individuals were infected during the 2001-2002 Havana DENV-3 epidemic. All but four sera collected from DF cases had a secondary DENV-1/DENV-3 infection. Sera neutralizing antibody titer against the seven DENV-3 strains were determined by plaque reduction neutralization technique. Sera samples were tested simultaneously. Studied sera showed higher levels of neutralizing antibodies to DENV-3 strains of genotype III compared to genotype V. Interesting, higher levels of neutralizing antibodies were detected to DENV-3 strain isolated at the end of the epidemic 2001-2002. An increased tendency of GMT of neutralizing antibodies according to epidemic evolution was observed for the 2001-2002 outbreak. In general, antibody levels in sera collected from DF cases were higher. Differences in the neutralization capacity of immune DENV-3 sera tested against two homologous genotypes including strains of the same genotype are demonstrated. Observed results suggest that virus changed in the course of the epidemic. The implications of this finding in terms of dengue pathogenesis and vaccine development need to be considered.

Secuencias de infección viral asociadas a la fiebre del dengue durante la epidemia de dengue 3 en la ciudad de La Habana, 2001-2002 / Viral infection sequences related to dengue fever in dengue 3 epidemics occurred in the City of Havana, 2001-2002

Objetivo: estudiar el papel de la infección secundaria y de la influencia de determinadas secuencias de infección virales en los casos de fiebre del dengue durante la epidemia de dengue 3, en la ciudad de La Habana, 2001-2002. Métodos: se estudiaron 141 muestras clínicas de casos confirmados de dengue en la epidemia cubana dengue 3. Todos los casos incluidos fueron clasificados de acuerdo con el criterio de la OMS como casos de fiebre del dengue, 101 de estas muestras fueron colectadas en la fase aguda de la enfermedad y 40 colectadas en la fase convaleciente tardía (16-18 meses después de la enfermedad). Resultados: los sueros colectados en la fase convaleciente tardía permitieron conocer las secuencias virales de infección, las cuales en orden descendiente fueron DEN-1/DEN-3, DEN-2/DEN-3 y DEN-1/DEN-2/DEN-3. Conclusiones: los resultados confirman que la secuencia de infección DEN-2/DEN-3 estuvo asociada a los casos de fiebre del dengue y no a los de fiebre hemorrágica del dengue; un porcentaje elevado de los casos estudiados se correspondió con una infección secundaria.

Objective: To study the role of secondary infection and of certain viral infection sequences in dengue fever cases during the dengue 3 epidemics occurred in the City of Havana. Methods: One hundred and forty one laboratory-confirmed clinical samples from dengue 3 cases were studied. According to WHO criteria, all included cases were classified as dengue fever cases; 101 of these samples were collected at the acute phase of the disease whereas 40 were collected in the late convalescence (16 to 18 months after the onset of disease). Results: The late convalescence serum samples allowed identifying the viral dengue infection sequences, which in downward order were DENV-1/DENV-3, DENV-2/DENV-3 and DENV-1/DENV-2/DENV-3. Conclusions: The results confirmed that the sequence infection DENV-2 / DENV-3 was associated with Dengue Fever Cases but not with the Dengue Hemorrhagic Fever Cases and that a high percentage of studied cases proved to be secondary infection.

Secuencias de infección viral asociadas a la fiebre del dengue durante la epidemia de dengue 3 en la ciudad de La Habana, 2001-2002 / Viral infection sequences related to dengue fever in dengue 3 epidemics occurred in the City of Havana, 2001-2002

Objetivo: estudiar el papel de la infección secundaria y de la influencia de determinadas secuencias de infección virales en los casos de fiebre del dengue durante la epidemia de dengue 3, en la ciudad de La Habana, 2001-2002. Métodos: se estudiaron 141 muestras clínicas de casos confirmados de dengue en la epidemia cubana dengue 3. Todos los casos incluidos fueron clasificados de acuerdo con el criterio de la OMS como casos de fiebre del dengue, 101 de estas muestras fueron colectadas en la fase aguda de la enfermedad y 40 colectadas en la fase convaleciente tardía (16-18 meses después de la enfermedad). Resultados: los sueros colectados en la fase convaleciente tardía permitieron conocer las secuencias virales de infección, las cuales en orden descendiente fueron DEN-1/DEN-3, DEN-2/DEN-3 y DEN-1/DEN-2/DEN-3. Conclusiones: los resultados confirman que la secuencia de infección DEN-2/DEN-3 estuvo asociada a los casos de fiebre del dengue y no a los de fiebre hemorrágica del dengue; un porcentaje elevado de los casos estudiados se correspondió con una infección secundaria(AU)

Objective: To study the role of secondary infection and of certain viral infection sequences in dengue fever cases during the dengue 3 epidemics occurred in the City of Havana. Methods: One hundred and forty one laboratory-confirmed clinical samples from dengue 3 cases were studied. According to WHO criteria, all included cases were classified as dengue fever cases; 101 of these samples were collected at the acute phase of the disease whereas 40 were collected in the late convalescence (16 to 18 months after the onset of disease). Results: The late convalescence serum samples allowed identifying the viral dengue infection sequences, which in downward order were DENV-1/DENV-3, DENV-2/DENV-3 and DENV-1/DENV-2/DENV-3. Conclusions: The results confirmed that the sequence infection DENV-2 / DENV-3 was associated with Dengue Fever Cases but not with the Dengue Hemorrhagic Fever Cases and that a high percentage of studied cases proved to be secondary infection(AU)

Primary and secondary infections of Macaca fascicularis monkeys with Asian and American genotypes of dengue virus 2

The goal of this study was to compare the immune response and the protection capacity induced by the dengue virus 2 (DENV-2) American and Asian genotypes in Macaca fascicularis monkeys. Animals were infected with American or Asian DENV-2 strains and challenged 1 year later with a DENV-2 Asian genotype strain. The viremia and monkey antibody levels were similar for the different strains after primary and secondary infection; however, the functionality of the antibody response was different. A limited viral replication was demonstrated after the secondary infection in all the monkeys. No virus was isolated in tissue culture, while reverse transcription-PCR showed a late positive reaction in four of five challenged monkeys. The immunoglobulin M response pattern and the detection of antibodies to specific proteins by Western blotting supported the protection data. Despite the demonstration of the protective effect after homologous challenge, a strong anamnestic antibody response was observed(AU)

Primary and secondary infections of Macaca fascicularis monkeys with Asian and American genotypes of dengue virus 2.

The goal of this study was to compare the immune response and the protection capacity induced by the dengue virus 2 (DENV-2) American and Asian genotypes in Macaca fascicularis monkeys. Animals were infected with American or Asian DENV-2 strains and challenged 1 year later with a DENV-2 Asian genotype strain. The viremia and monkey antibody levels were similar for the different strains after primary and secondary infection; however, the functionality of the antibody response was different. A limited viral replication was demonstrated after the secondary infection in all the monkeys. No virus was isolated in tissue culture, while reverse transcription-PCR showed a late positive reaction in four of five challenged monkeys. The immunoglobulin M response pattern and the detection of antibodies to specific proteins by Western blotting supported the protection data. Despite the demonstration of the protective effect after homologous challenge, a strong anamnestic antibody response was observed.

Caracterización microbiológica de cepas de Vibrio cholerae no-O1 aisladas en Cuba / Microbiological characterization of non-01 Vibrio cholerae isolated in Cuba

Se estudiaron 422 cepas de Vibrio cholerae no-O1 procedentes de 9 provincias del país, de ellas 9 aisladas de un brote de enfermedad de transmisión hídrica. En la totalidad de las cepas se determinó la susceptibilidad antimicrobiana y la presencia de factores de virulencia. En las 9 cepas procedentes del brote, se realizó además, el estudio de macrorrestricción de ADN mediante la técnica de electroforesis de campo pulsado. Se demostró por primera vez en Cuba y el Caribe, la circulación de cepas de V. cholerae no-O1 atípicas (resistentes al compuesto vibriostático O129 y al trimetoprim-sulfametoxazol). El comportamiento de la susceptibilidad antimicrobiana demostró por primera vez la circulación en Cuba de 2 patrones diferentes de resistencia (ampicilina, trimetoprim/sulfametoxazol, sulfonamida y tetraciclina, trimetoprim/sulfametoxazol, sulfonamida). La frecuencia de cepas resistentes al trimetoprim-sulfametoxazol se mantuvo similar en todo el período de estudio. Sin embargo, se produjo un decrecimiento de la resistencia a la ampicilina y un aumento de la resistencia a la tetraciclina. Los principales factores de virulencia que se encontraron fueron la gelatinasa, la hemolisina, la elastasa y la adherencia a células HEp-2. Sin embargo, las cepas del brote mostraron mayores porcentajes que el resto, para la presencia de la toxina termoestable y la presencia de fimbrias. Los resultados de los estudios molecular y epidemiológico permitieron dar una respuesta acelerada y precisa sobre la etiología del primer brote de enfermedad de transmisión alimentaria.

The study of 422 non-01 Vibrio cholerae strains from nine provinces, 9 of them isolated from a water-borne disease outbreak, was performed. All the strains exhibited antimicrobial susceptibility and virulence factors. The nine strains from the outbreak were subjected to a DNA macrorestriction study based on the pulsed field electrophoresis technique. For the first time in Cuba and the Caribbean, the circulation of atypical non-01 V. Cholerae strains (resistent to vibriostatic compound 0129 and trimethoprim/sulfamethoxazole). The behavior of antimicrobial susceptibility evinced for the first time the circulation of two different resistence patterns in Cuba (ampicilline, trimethoprim/sulfamethoxazole, sulfonamide and tetracycline, trimethoprim/sulfamethoxazole, sulfonamide). The frequency of trimethoprim/sulfamethoxazole-resistent strains was similar during the whole period of study. However, resistance to ampicilline decreased whereas resistance to tetracycline increased. The main found virulence factors were gelatinase, hemolysine, elastase and adherence to Hep-2 cells. On the other hand, the outbreak strains showed higher percentages than the others due to the presence of heat-liable toxin and fimbriae. The results of the molecular and epidemiological studies allowed giving a speedy and accurate response that explained the etiology of the first food-borne disease outbreak.

Caracterización microbiológica de cepas de Vibrio cholerae no-O1 aisladas en Cuba / Microbiological characterization of non-01 Vibrio cholerae isolated in Cuba

Se estudiaron 422 cepas de Vibrio cholerae no-O1 procedentes de 9 provincias del país, de ellas 9 aisladas de un brote de enfermedad de transmisión hídrica. En la totalidad de las cepas se determinó la susceptibilidad antimicrobiana y la presencia de factores de virulencia. En las 9 cepas procedentes del brote, se realizó además, el estudio de macrorrestricción de ADN mediante la técnica de electroforesis de campo pulsado. Se demostró por primera vez en Cuba y el Caribe, la circulación de cepas de V. cholerae no-O1 atípicas (resistentes al compuesto vibriostático O129 y al trimetoprim-sulfametoxazol). El comportamiento de la susceptibilidad antimicrobiana demostró por primera vez la circulación en Cuba de 2 patrones diferentes de resistencia (ampicilina, trimetoprim/sulfametoxazol, sulfonamida y tetraciclina, trimetoprim/sulfametoxazol, sulfonamida). La frecuencia de cepas resistentes al trimetoprim-sulfametoxazol se mantuvo similar en todo el período de estudio. Sin embargo, se produjo un decrecimiento de la resistencia a la ampicilina y un aumento de la resistencia a la tetraciclina. Los principales factores de virulencia que se encontraron fueron la gelatinasa, la hemolisina, la elastasa y la adherencia a células HEp-2. Sin embargo, las cepas del brote mostraron mayores porcentajes que el resto, para la presencia de la toxina termoestable y la presencia de fimbrias. Los resultados de los estudios molecular y epidemiológico permitieron dar una respuesta acelerada y precisa sobre la etiología del primer brote de enfermedad de transmisión alimentaria(AU)

The study of 422 non-01 Vibrio cholerae strains from nine provinces, 9 of them isolated from a water-borne disease outbreak, was performed. All the strains exhibited antimicrobial susceptibility and virulence factors. The nine strains from the outbreak were subjected to a DNA macrorestriction study based on the pulsed field electrophoresis technique. For the first time in Cuba and the Caribbean, the circulation of atypical non-01 V. Cholerae strains (resistent to vibriostatic compound 0129 and trimethoprim/sulfamethoxazole). The behavior of antimicrobial susceptibility evinced for the first time the circulation of two different resistence patterns in Cuba (ampicilline, trimethoprim/sulfamethoxazole, sulfonamide and tetracycline, trimethoprim/sulfamethoxazole, sulfonamide). The frequency of trimethoprim/sulfamethoxazole-resistent strains was similar during the whole period of study. However, resistance to ampicilline decreased whereas resistance to tetracycline increased. The main found virulence factors were gelatinase, hemolysine, elastase and adherence to Hep-2 cells. On the other hand, the outbreak strains showed higher percentages than the others due to the presence of heat-liable toxin and fimbriae. The results of the molecular and epidemiological studies allowed giving a speedy and accurate response that explained the etiology of the first food-borne disease outbreak(AU)

Neutralizing antibodies after infection with dengue 1 virus.

Severity of disease is markedly increased when infection with dengue virus type 2 (DENV-2) follows infection with DENV-1 at an interval of 20 years. Studies have shown that heterologous neutralizing antibody titers are inversely correlated with severity of a second infection. If this mechanism controlled disease severity in Cuba, heterotypic antibody titers should have declined over time. To determine whether phenotypic changes in dengue antibodies occur over time, we analyzed serum samples collected 4-8 and 20-22 years after DENV-1 infection. We found a significant increase in mean titer of homologous DENV-1 neutralizing antibodies and a significant decrease in heterologous antibodies to 1 of 2 genotypes of DENV-2 virus (the American genotype). Asian DENV-2 viruses were not neutralized during either interval; however, the American genotype underwent phenotypic changes in heterotypic viral neutralizing antibodies in the predicted direction. This finding may be related to the time-dependent changes in severity of disease found with secondary dengue infection.

RETRACTED: Evidence for Other Non-Poliovirus Enteroviruses Multiplies in L20B Cells.

This article has been retracted.