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

Molecular Phylogeny and Proposed Classification of the Simian Picornaviruses

The simian picornaviruses were isolated from various primate tissues during the development of general tissue culture methods in the 1950s to 1970s or from specimens derived from primates used in biomedical research. Twenty simian picornavirus serotypes are recognized, and all are presently classified within the Enterovirus genus. To determine the phylogenetic relationships among all of the simian picornaviruses and to evaluate their classification, we have determined complete VP1 sequences for 19 of the 20 serotypes. Phylogenetic analysis showed that A13, SV19, SV26, SV35, SV43, and SV46 are members of human enterovirus species A, a group that contains enterovirus 71 and 11 of the coxsackie A viruses. SA5 is a member of human enterovirus species B, which contains the echoviruses, coxsackie B viruses, coxsackievirus A9, and enterovirus 69. SV6, N125, and N203 are related to one another and, more distantly, to species A human enteroviruses, but could not be definitely assigned to a species. SV4 and SV28 are closely related to one another and to A-2 plaque virus, but distinct from other enteroviruses, suggesting that these simian viruses are members of a new enterovirus species. SV2, SV16, SV18, SV42, SV44, SV45, and SV49 are related to one another but distinct from viruses in all other picornavirus genera, suggesting that they may comprise a previously unknown genus in Picornaviridae. Several simian virus VP1 sequences (N125 and N203; SV4 and SV28; SV19, SV26, and SV35; SV18 and SV44; SV16, SV42, and SV45) are greater than 75% identical to one another (and/or greater than 85% amino acid identity), suggesting that the true number of distinct serotypes among the viruses surveyed is less than 20

Evidence for Frequent Recombination within Species Human Enterovirus B Based on Complete Genomic Sequences of All Thirty-Seven Serotypes

The species Human enterovirus B (HEV-B) in the family Picornaviridae consists of coxsackievirus A9; coxsackieviruses B1 to B6; echoviruses 1 to 7, 9, 11 to 21, 24 to 27, and 29 to 33; and enteroviruses 69 and 73. We have determined complete genome sequences for the remaining 22 HEV-B serotypes whose sequences were not represented in public databases and analyzed these in conjunction with previously available complete sequences in GenBank. Members of HEV-B were monophyletic relative to all other human enterovirus species in all regions of the genome except in the 5′-nontranslated region (NTR), where they are known to cluster with members of HEV-A. Within HEV-B, phylogenies constructed from the structural (P1) and nonstructural regions of the genome (P2 and P3) are incongruent, suggesting that recombination had occurred. Similarity plots and bootscanning analysis across the complete genome identified multiple sites at which the phylogeny of a given strain's sequence shifted, indicating potential recombination points. These points are distributed in the 5′-NTR and throughout P2 and P3, but no sites with >80% bootstrap support were identified within the capsid. Individual sequence comparisons and phylogenetic analyses suggest that members of HEV-B have recombined with one another on multiple occasions, resulting in a complex mosaic of sequences derived from multiple parental viruses in the nonstructural regions of the genome. We conclude that RNA recombination is a common mechanism for enterovirus evolution and that recombination within the nonstructural regions of the genome (P2 and P3) has been observed only among members of the same species

Identification of Enteroviruses in Naturally Infected Captive Primates▿

In a recent study, we investigated cases of diarrheal disease among monkeys at a U.S. primate center. In that study, enteroviruses were detected in a high proportion of the fecal specimens tested. To determine whether the enterovirus detections represented the circulation of one or more simian enteroviruses within the colony or the transmission of human enteroviruses from animal handlers, we determined in the present study the serotype identity of each virus by reverse transcription-PCR and sequencing of a portion of the VP1 gene, a region whose sequence corresponds to antigenic type. Enteroviruses were identified in 37 of 56 specimens (66%), 30 of 40 rhesus macaques, 5 of 11 pigtail macaques, 2 of 4 sooty mangabeys, and 0 of 1 chimpanzee. No previously known human viruses were detected. Three previously known simian enterovirus serotypes—SV6, SV19, and SV46—were among the viruses identified, but more than half of the identified viruses were previously unknown; these have been assigned as new types: EV92 and EV103

Detection of All Known Parechoviruses by Real-Time PCR▿

The Parechovirus genus of the Picornaviridae family contains two species, Human parechovirus (HPeV) and Ljungan virus (LV). The HPeVs (including the former echoviruses 22 and 23, now HPeV type 1 (HPeV1) and HPeV2, respectively) cause a wide spectrum of disease, including aseptic meningitis, gastroenteritis, encephalitis, acute respiratory illness, and neonatal sepsis-like disease. The LVs were isolated from bank voles in Sweden during a search for an infectious agent linked to fatal myocarditis cases in humans. Because of the decline in use of cell culture and neutralization to investigate enterovirus-like disease, very few laboratories currently have the capability to test for parechoviruses. We have developed a real-time reverse transcription-PCR (RT-PCR) assay for detection of all known members of the genus Parechovirus. The assay targets the conserved regions in the 5′ nontranslated region (5′NTR) of the parechovirus genome and can detect both HPeVs and LVs, unlike other published parechovirus 5′ NTR assays, which only detect known HPeVs or only LVs. HPeV and LV can be differentiated by sequencing the 5′NTR real-time RT-PCR amplicon, when needed. The assay is approximately 100 times more sensitive than cell culture and may be used to test original clinical specimens. The availability of a broad-specificity PCR method should facilitate the detection of new human parechoviruses, as well as new parechoviruses in other mammalian species, and provide an opportunity to investigate the role of these viruses in human and animal disease