Molecular identification and characterization of two proposed new enterovirus serotypes, EV74 and EV75

Fil: Oberste, M. Steven. Centers for Disease Control and Prevention. Respiratory and Enteric Viruses Branch; Estados Unidos.Fil: Michele, Suzanne M. Centers for Disease Control and Prevention. Respiratory and Enteric Viruses Branch; Estados Unidos.Fil: Maher, Kaija. Centers for Disease Control and Prevention. Respiratory and Enteric Viruses Branch; Estados Unidos.Fil: Schnurr, David. California Department of Health Services. Viral and Rickettsial Disease Laboratory; Estados Unidos.Fil: Cisterna, Daniel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Junttila, Nina. Swedish Institute for Disease Control. Department of Virology; Suecia.Fil: Uddin, Moyez. Institute of Public Health; Bangladesh.Fil: Chomel, Jean-Jacques. Centre National de Référence des Entérovirus; Francia.Fil: Lau, Chi-Shan. Queen Mary Hospital. Department of Health; China.Fil: Ridha, Walid. National Polio Laboratory; Irak.Fil: Al-Busaidy, Suleiman. Ministry of Health. Department of Laboratories; Oman.Fil: Norder, Helene. Swedish Institute for Disease Control. Department of Virology; Suecia.Fil: Magnius, Lars O. Swedish Institute for Disease Control. Department of Virology; Suecia.Fil: Pallansch, Mark A. Centers for Disease Control and Prevention. Respiratory and Enteric Viruses Branch; Estados Unidos.Sequencing of the gene that encodes the capsid protein VP1 has been used as a surrogate for antigenic typing in order to distinguish enterovirus serotypes; three new serotypes were identified recently by this method. In this study, 14 enterovirus isolates from six countries were characterized as members of two new types within the species Human enterovirus B, based on sequencing of the complete capsid-encoding (P1) region. Isolates within each of these two types differed significantly from one another and from all other known enterovirus serotypes on the basis of sequences that encode either VP1 alone or the entire P1 region. Members of each type were greater than or equal to 77(.)2% identical to one another (89(.)5% amino acid identity) in VP1, but members of the two different types differed from one another and from other enteroviruses by greater than or equal to 31% in nucleotide sequence (25% amino acid sequence difference), indicating that the two groups represent separate new candidate enterovirus types. The complete P1 sequences differed from those of all other enterovirus serotypes by greater than or equal to 31% (26% amino acid sequence difference), but were highly conserved within a serotype (< 8% amino acid sequence difference). Phylogenetic analyses demonstrated that isolates of the same serotype were monophyletic in both VP1 and the capsid as a whole, as shown previously for other enterovirus serotypes. This paper proposes that these 14 isolates should be classified as members of two new human enterovirus types, enteroviruses 74 and 75 (EV74 and EV75)

Diversity of picornaviruses in rural Bolivia

Fil: Nix, W Allan. Centers for Disease Control and Prevention; Estados Unidos.Fil: Khetsuriani, Nino. Centers for Disease Control and Prevention; Estados Unidos.Fil: Peñaranda, Silvia. Centers for Disease Control and Prevention; Estados Unidos.Fil: Maher, Kaija. Centers for Disease Control and Prevention; Estados Unidos.Fil: Venczel, Linda. Centers for Disease Control and Prevention; Estados Unidos.Fil: Cselkó, Zsuzsa. Centers for Disease Control and Prevention; Estados Unidos.Fil: Freire, Maria Cecilia. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Cisterna, Daniel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Lema, Cristina L. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Rosales, Patricia. Ministry of Health and Sports; Bolivia.Fil: Rodríguez, Jacqueline R. Pediatric Hospital San Antonio de Los Sauces; Bolivia.Fil: Rodríguez, Wilma. Ministry of Health and Sports; Bolivia.Fil: Halkyer, Percy. Pan-American Health Organization; Bolivia.Fil: Ronveaux, Olivier. Pan-American Health Organization; Bolivia.Fil: Pallansch, Mark A. Centers for Disease Control and Prevention; Estados Unidos.Fil: Oberste, M Steven. Centers for Disease Control and Prevention; Estados Unidos.The family Picornaviridae is a large and diverse group of viruses that infect humans and animals. Picornaviruses are among the most common infections of humans and cause a wide spectrum of acute human disease. This study began as an investigation of acute flaccid paralysis (AFP) in a small area of eastern Bolivia, where surveillance had identified a persistently high AFP rate in children. Stools were collected and diagnostic studies ruled out poliovirus. We tested stool specimens from 51 AFP cases and 34 healthy household or community contacts collected during 2002-2003 using real-time and semi-nested reverse transcription polymerase chain reaction assays for enterovirus, parechovirus, cardiovirus, kobuvirus, salivirus and cosavirus. Anecdotal reports suggested a temporal association with neurological disease in domestic pigs, so six porcine stools were also collected and tested with the same set of assays, with the addition of an assay for porcine teschovirus. A total of 126 picornaviruses were detected in 73 of 85 human individuals, consisting of 53 different picornavirus types encompassing five genera (all except Kobuvirus). All six porcine stools contained porcine and/or human picornaviruses. No single virus, or combination of viruses, specifically correlated with AFP; however, the study revealed a surprising complexity of enteric picornaviruses in a single community

Detection of Respiratory Viruses and Subtype Identification of Influenza A Viruses by GreeneChipResp Oligonucleotide Microarray

Acute respiratory infections are significant causes of morbidity, mortality, and economic burden worldwide. An accurate, early differential diagnosis may alter individual clinical management as well as facilitate the recognition of outbreaks that have implications for public health. Here we report on the establishment and validation of a comprehensive and sensitive microarray system for detection of respiratory viruses and subtyping of influenza viruses in clinical materials. Implementation of a set of influenza virus enrichment primers facilitated subtyping of influenza A viruses through the differential recognition of hemagglutinins 1 through 16 and neuraminidases 1 through 9. Twenty-one different respiratory virus species were accurately characterized, including a recently identified novel genetic clade of rhinovirus.Fil: Quan, Phenix-Lan. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Palacios, Gustavo. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Jabado, Omar J. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Conlan, Sean. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Hirschberg, David L. Stanford School of Medicine; Estados Unidos.Fil: Pozo, Francisco. Instituto de Salud Carlos III. Centro Nacional de Microbiología; España.Fil: Jack, Philippa J. M. Australian Animal Health Laboratory. CSIRO Livestock Industries; Australia.Fil: Cisterna, Daniel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Renwick, Neil. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Hui, Jeffrey. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Drysdale, Andrew. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Amos-Ritchie, Rachel. Australian Animal Health Laboratory. CSIRO Livestock Industries; Australia.Fil: Baumeister, Elsa. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Savy, Vilma. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Lager, Kelly M. USDA. National Animal Disease Center; Estados Unidos.Fil: Richt, Jürgen A. USDA. National Animal Disease Center; Estados Unidos.Fil: Boyle, David B. Australian Animal Health Laboratory. CSIRO Livestock Industries; Australia.Fil: García-Sastre, Adolfo. Mount Sinai School of Medicine. Department of Microbiology and Emerging Pathogens Institute; Estados Unidos.Fil: Casas, Inmaculada. Instituto de Salud Carlos III. Centro Nacional de Microbiología; España.Fil: Perez-Breña, Pilar. Instituto de Salud Carlos III. Centro Nacional de Microbiología; España.Fil: Briese, Thomas. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Lipkin, W. Ian. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos

Detection of Respiratory Viruses and Subtype Identification of Influenza A Viruses by GreeneChipResp Oligonucleotide Microarray

Acute respiratory infections are significant causes of morbidity, mortality, and economic burden worldwide. An accurate, early differential diagnosis may alter individual clinical management as well as facilitate the recognition of outbreaks that have implications for public health. Here we report on the establishment and validation of a comprehensive and sensitive microarray system for detection of respiratory viruses and subtyping of influenza viruses in clinical materials. Implementation of a set of influenza virus enrichment primers facilitated subtyping of influenza A viruses through the differential recognition of hemagglutinins 1 through 16 and neuraminidases 1 through 9. Twenty-one different respiratory virus species were accurately characterized, including a recently identified novel genetic clade of rhinovirus.Fil: Quan, Phenix-Lan. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Palacios, Gustavo. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Jabado, Omar J. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Conlan, Sean. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Hirschberg, David L. Stanford School of Medicine; Estados Unidos.Fil: Pozo, Francisco. Instituto de Salud Carlos III. Centro Nacional de Microbiología; España.Fil: Jack, Philippa J. M. Australian Animal Health Laboratory. CSIRO Livestock Industries; Australia.Fil: Cisterna, Daniel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Renwick, Neil. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Hui, Jeffrey. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Drysdale, Andrew. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Amos-Ritchie, Rachel. Australian Animal Health Laboratory. CSIRO Livestock Industries; Australia.Fil: Baumeister, Elsa. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Savy, Vilma. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas; Argentina.Fil: Lager, Kelly M. USDA. National Animal Disease Center; Estados Unidos.Fil: Richt, Jürgen A. USDA. National Animal Disease Center; Estados Unidos.Fil: Boyle, David B. Australian Animal Health Laboratory. CSIRO Livestock Industries; Australia.Fil: García-Sastre, Adolfo. Mount Sinai School of Medicine. Department of Microbiology and Emerging Pathogens Institute; Estados Unidos.Fil: Casas, Inmaculada. Instituto de Salud Carlos III. Centro Nacional de Microbiología; España.Fil: Perez-Breña, Pilar. Instituto de Salud Carlos III. Centro Nacional de Microbiología; España.Fil: Briese, Thomas. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos.Fil: Lipkin, W. Ian. Columbia University. Jerome L. and Dawn Greene Infectious Disease Laboratory; Estados Unidos

Whole genome sequencing identifies independent outbreaks of Shigellosis in 2010 and 2011 in La Pampa Province, Argentina

AbstractShigella sonnei is an emergent cause of diarrheal disease in middle-income countries. The organism causes endemic disease and is also associated with sporadic outbreaks in susceptible populations. In 2010 and 2011 there were two suspected outbreaks of diarrheal disease caused by S. sonnei in La Pampa province in central Argentina. Aiming to confirm these as outbreaks and provide insight into the relationship of the strains causing these infections we combined antimicrobial susceptibility testing and pulsed field gel electrophoresis (PFGE) with whole genome sequencing (WGS). Antimicrobial susceptibility testing suggested the two events were unrelated; organisms isolated in 2010 exhibited resistance to trimethoprim sulphate whereas the 2011 S. sonnei were non-susceptible against ampicillin, trimethoprim sulphate and cefpodoxime. PFGE profiling confirmed the likelihood of two independent outbreaks, separating the isolates into two main XbaI restriction profiles. We additionally performed WGS on 17 isolates associated with these outbreaks. The resulting phylogeny confirmed the PFGE structure and separated the organisms into two comparatively distantly related clones. Antimicrobial resistant genes were common, and the presence of an OXA-1 was likely associated with resistance to cefpodoxime in the second outbreak. We additionally identified novel horizontally transferred genetic material that may impinge on the pathogenic phenotype of the infecting strains. Our study shows that even with a lack of supporting routine data WGS is an indispensible method for the tracking and surveillance of bacterial pathogens during outbreaks and is becoming a vital tool for the monitoring of antimicrobial resistant strains of S. sonnei.</jats:p

Outbreak of viral meningitis caused by echovirus type 4 in Misiones province

Fil: Grenbon, S. L. Universidad Nacional de Misiones. Hospital Provincial de Pediatría; Argentina.Fil: Robledo, M. L. Universidad Nacional de Misiones. Hospital Provincial de Pediatría; Argentina.Fil: von Specht, M. H. Universidad Nacional de Misiones. Hospital Provincial de Pediatría; Argentina.Fil: Cisterna, Daniel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología. Servicio de Neurovirosis; Argentina.Fil: Lema, C. L. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología. Servicio de Neurovirosis; Argentina.Fil: Freire, María Cecilia. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Virología. Servicio de Neurovirosis; Argentina.Se realizó un estudio retrospectivo a fin de describir un brote epidémico de meningitis causado por enterovirus, que comprometió a 143 niños de 1 mes a 14 años internados en el Hospital Pediátrico de Posadas (Misiones) con diagnóstico de meningitis aséptica, entre agosto y diciembre de 2005. Se observó un aumento de casos entre las semanas 33 a 50, con un pico máximo entre las semanas 47 y 48, lo que confirmó el brote. La mediana de edad de los niños afectados fue de 8 años y el 55,2% fueron varones. El 80% de los casos se observó entre escolares (5 a 14 años). El promedio del tiempo de internación fue de 4,5 ± 1,7 días, y no se registraron fallecidos. Los LCR se estudiaron mediante examen citoquímico y estudios bacteriológicos y virológicos (aislamiento viral, RT- PCR anidada e identificación molecular mediante secuenciación génica). Los recuentos de células en LCR variaron entre 6 y 5040 células /mm3, el 92% fueron inferiores a 500 células/mm3 y el 43,5% mostró predominio linfocitario. El 56% presentó concentraciones de glucosa normal, con proteínas ligeramente elevadas. El 28% de las muestras estudiadas por cultivo (17/60) mostró efecto citopático, compatible con enterovirus. La RT-PCR anidada permitió detectar enterovirus en un 73% de las muestras (43/59), con 6 casos que se tipificaron como echovirus tipo 4. El índice de positividad al combinar ambas técnicas alcanzó el 83%