ABSTRACT
Human metapneumovirus (hMPV) has been described as circulating among the Uruguayan population at least since 1998 based on serologic evidence. However, no isolation attempts, molecular detection, or genetic studies have been carried out so far in the country. In the present study, molecular detection of circulating hMPV in children hospitalized with acute respiratory tract infection in Montevideo-Uruguay was carried out by reverse transcription-polymerase chain reaction (RT-PCR) amplification of the hMPV nucleoprotein (N) gene from 217 nasopharyngeal aspirates. Genetic variability analysis of the positive samples was performed by amplification and sequencing of both N and attachment glycoprotein (G) genes. Eighteen of the 217 samples tested positive for hMPV, with tachypnea, chest indrawing, and wheezing being the main clinical symptoms recorded. Phylogenetic analysis of N and G genes showed that Uruguayan samples clustered in genotypes described previously as A2, B1, and B2, with bootstrap values >or=98%. Sublineages A2a and A2b could also be distinguished within the samples that belong to A2. This is the first molecular report on the circulation of hMPV in Uruguay. The pattern of circulation of this virus, analyzed for both N and G genes independently, resembles the complex evolutionary pattern of respiratory syncytial virus (RSV).
Subject(s)
Genetic Variation , Metapneumovirus/classification , Metapneumovirus/isolation & purification , Paramyxoviridae Infections/virology , Cluster Analysis , Female , Humans , Infant , Infant, Newborn , Male , Metapneumovirus/genetics , Molecular Epidemiology , Molecular Sequence Data , Nasopharynx/virology , Nucleoproteins/genetics , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Uruguay , Viral Structural Proteins/geneticsABSTRACT
Adenoviruses are one of the most frequent causative agents of acute lower respiratory infections in infants and young children. Twenty-three adenovirus isolates from nasopharyngeal aspirates of children hospitalized for acute lower respiratory infections in Uruguay between 1994 and 1998 were studied by restriction enzyme analysis. The genomic analysis showed that 60.9% (n = 14) of isolates belonged to the species Human adenovirus C (HAdV-C) and 31.9% (n = 9) to the species Human adenovirus B (HAdV-B). Whereas some isolates could be classified according to the published profiles into genotype or genomic variant, others displayed migration patterns not allowing classification. Eight isolates (89%) of HAdV-B corresponded to the Ad7h genotype that has been associated with severe and fatal pneumonia and necrotizing bronchiolitis in children in South America. The isolates of HAdV-C showed a great variability in accordance with the data published earlier.
Subject(s)
Adenovirus Infections, Human/virology , Adenoviruses, Human/classification , Adenoviruses, Human/genetics , Respiratory Tract Infections/virology , Adenoviruses, Human/isolation & purification , Child, Preschool , DNA, Viral/genetics , DNA, Viral/isolation & purification , Genotype , Humans , Nasopharynx/virology , Restriction Mapping , UruguayABSTRACT
Genotypes of Human respiratory syncytial virus (HRSV) of group B from Uruguay were assigned to strains isolated during 1999 and 2001 outbreaks and others formerly reported isolated in the period 1989-1996. The nucleotide sequences of the C-terminal portion of the G protein were compared to sequences representative of previously defined HRSV genotypes. Most Uruguayan strains clustered into five of the previously identified genotypes. Nine isolates clustered in two genotypes named URU1 and URU2 which were not described up to present. Two of the analyzed sequences isolated in 2001 have a six nucleotide duplication that is discussed in terms of HRSV variability.
Subject(s)
Disease Outbreaks , Respiratory Syncytial Virus Infections/epidemiology , Respiratory Syncytial Virus, Human/genetics , Genotype , Humans , Molecular Epidemiology , Molecular Sequence Data , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Virus, Human/isolation & purification , Uruguay/epidemiologyABSTRACT
The genetic and antigenic variability of the G glycoproteins from 76 human respiratory syncytial (RS) viruses (subgroup A) isolated during six consecutive epidemics in either Montevideo, Uruguay, or Madrid, Spain, have been analyzed. Genetic diversity was evaluated for all viruses by the RNase A mismatch cleavage method and for selected strains by dideoxy sequencing. The sequences reported here were added to those published for six isolates from Birmingham, United Kingdom, and for two reference strains (A2 and Long), to derive a phylogenetic tree of subgroup A viruses that contained two main branches and several subbranches. During the same epidemic, viruses from different branches were isolated. In addition, closely related viruses were isolated in distant places and in different years. These results illustrate the capacity of the virus to spread worldwide, influencing its mode of evolution. The antigenic analysis of all isolates was carried out with a panel of anti-G monoclonal antibodies that recognized strain-specific (or variable) epitopes. A close correlation between genetic relatedness and antigenic relatedness in the G protein was observed. These results, together with an accumulation of amino acid changes in a major antigenic area of the G glycoprotein, suggest that immune selection may be a factor influencing the generation of RS virus diversity. The pattern of RS virus evolution is thus similar to that described for influenza type B viruses, expect that the level of genetic divergence among the G glycoproteins of RS virus isolates is the highest reported for an RNA virus gene product.
Subject(s)
HN Protein , Respiratory Syncytial Virus Infections/epidemiology , Respiratory Syncytial Viruses/genetics , Viral Proteins/genetics , Amino Acid Sequence , Antigenic Variation , Base Sequence , Biological Evolution , DNA Primers/chemistry , Disease Outbreaks , Genes, Viral , Humans , Molecular Sequence Data , Phylogeny , Respiratory Syncytial Viruses/immunology , Sequence Alignment , Sequence Homology, Amino Acid , Spain , Uruguay , Viral Envelope Proteins , Viral Fusion Proteins/genetics , Viral Proteins/immunology , Viral Structural Proteins/geneticsABSTRACT
El empleo simultáneo de técnicas de diagnóstico para bacterias, virus, Mycoplasma pneumoniae y Chlamydia trachomatis permitió identificar infecciones mixtas en procesos respiratorios agudos de niños menores de 5 años. Se logró reconocer la frecuencia relativa de estas infecciones y el porcentaje de los diferentes agentes implicados en la etiología de la neumonía adquirida en la comunidad así como en la otitis media aguda supurada. Entre 1987 y 1990 se captaron 541 pacientes con neumonía, determinándose la etiología en 63.7 por ciento de los casos. De esos 9.3 por ciento correspondió a una infección mixta. La mayor frecuencia relativa demostrada correspondió al aislamiento de una bacteria de la sangre y a la presencia de un virus en el aspirado nasofaringeo. En segundo lugar lo ocuparon las dobles infecciones virales. A pesar de que el virus respiratorio sincicial es por lejos el virus más frecuente, proporcionalmente se asoció menos a una bacteria que los adenovirus o los virus parainfluenza. De 62 exudados ópticos procesados, se diagnosticó un patógeno en 50 (80.6 por ciento). En 34 por ciento de las etiologías se asociaron 2 o más agentes. Se pudo concluir que las infecciones concomitantes por más de un patógeno ocurren no sólo en inmunodeprimidos sino que se hallan implicados en procesos respiratorios agudos de población general, y que su reconocimiento se logra empleando simultáneamente diversas técnicas y disponiendo de reactivos que posibiliten cubrir un espectro amplio de diferentes patógenos respiratorios
Subject(s)
Humans , Male , Female , Pregnancy , Infant, Newborn , Infant , Child, Preschool , Infections , Otitis Media , Pneumonia , Pneumonia/diagnosis , Pneumonia/etiology , Otitis Media/etiologyABSTRACT
El empleo simultáneo de técnicas de diagnóstico para bacterias, virus, Mycoplasma pneumoniae y Chlamydia trachomatis permitió identificar infecciones mixtas en procesos respiratorios agudos de niños menores de 5 años. Se logró reconocer la frecuencia relativa de estas infecciones y el porcentaje de los diferentes agentes implicados en la etiología de la neumonía adquirida en la comunidad así como en la otitis media aguda supurada. Entre 1987 y 1990 se captaron 541 pacientes con neumonía, determinándose la etiología en 63.7 por ciento de los casos. De esos 9.3 por ciento correspondió a una infección mixta. La mayor frecuencia relativa demostrada correspondió al aislamiento de una bacteria de la sangre y a la presencia de un virus en el aspirado nasofaringeo. En segundo lugar lo ocuparon las dobles infecciones virales. A pesar de que el virus respiratorio sincicial es por lejos el virus más frecuente, proporcionalmente se asoció menos a una bacteria que los adenovirus o los virus parainfluenza. De 62 exudados ópticos procesados, se diagnosticó un patógeno en 50 (80.6 por ciento). En 34 por ciento de las etiologías se asociaron 2 o más agentes. Se pudo concluir que las infecciones concomitantes por más de un patógeno ocurren no sólo en inmunodeprimidos sino que se hallan implicados en procesos respiratorios agudos de población general, y que su reconocimiento se logra empleando simultáneamente diversas técnicas y disponiendo de reactivos que posibiliten cubrir un espectro amplio de diferentes patógenos respiratorios (AU)