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...gene (270 bp)sof 221 sequences using ClustalW in the Bioedit version 7.0.9 (17). For determination of RSV group Asgenotypes, phylogenetic analyses were performed using the PHYLIP version 3.64 package =-=(11)-=-.sSequence distances were calculated from alignments using F84, and clustering was done by thesneighbor-joining algorithm of the PHYLIP program package. Unique representative sequences of eachsof the ...

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...lied Biosystems).sPhylogenetic analysis.sA multiple-sequence alignment was compiled from the second variable region of the G gene (270 bp)sof 221 sequences using ClustalW in the Bioedit version 7.0.9 =-=(17)-=-. For determination of RSV group Asgenotypes, phylogenetic analyses were performed using the PHYLIP version 3.64 package (11).sSequence distances were calculated from alignments using F84, and cluster...

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... which is nowsrepresented as a subtype in the tree. Evaluation of the robustness of the tree was performed bysbootstrap analysis carried out with 1,000 replicates. The tree was plotted using TREEVIEW =-=(33)-=- andswas manually edited in Corel Draw 12 program.sComparisons between nucleotide or amino acid sequences were calculated using the DNADist orsProtdist tool, respectively, of the Bioedit software, and...

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...mbers of potentially Oglycosylated serine and threonine residues, NetOglyc software (version 3.1) (18) was used.sSynonymous and nonsynonymous mutations were analyzed by the method of Nei and Gojobori =-=(31)-=-.sThe program SNAP (Synonymous/Nonsynonymous Analysis Program) provided by the humansimmunodeficiency virus (HIV) sequence database websites(http://www.hiv.lanl.gov/content/sequence/SNAP/SNAP.html) wa...

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...virus to infect people repeatedly and cause yearly outbreaks (38). RSV has a clearsseasonality. In temperate climates, outbreaks occur yearly in the late fall, winter, or spring but not insthe summer =-=(22, 29, 61)-=-. Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics (2, 4, 13, 20), although group A viruses tend to predominates(8).sThere is limited i...

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... in the extracellular domain. Diversity occurs mainly in the Gsectodomain which has only 44% amino acid identity between the two groups compared with 83% forsthe transmembrane and cytoplasmic domains =-=(21)-=-. The G protein is heavily glycosylated with N-linkedsand O-linked sugars (60). Mainly, the ectodomain of the G protein has a high content of serine andsthreonine residues which are potential acceptor...

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... immunity (19).sRSV strains are separated into two major groups based on antigenic and genetic variability. The mainsdifferences between RSV groups A and B were found in the attachment glycoprotein G =-=(1, 6, 30)-=-.sVariability in this protein is greater than that in the other proteins, both between and within the majorsantigenic groups of RSV. The G protein is a type II glycoprotein of 289 to 299 amino acids i...

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... immunity (19).sRSV strains are separated into two major groups based on antigenic and genetic variability. The mainsdifferences between RSV groups A and B were found in the attachment glycoprotein G =-=(1, 6, 30)-=-.sVariability in this protein is greater than that in the other proteins, both between and within the majorsantigenic groups of RSV. The G protein is a type II glycoprotein of 289 to 299 amino acids i...

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...wer respiratory tractsinfections in infants and young children in both developing and developed countries (46). By the age ofs2 years, virtually all children have been infected at least once with RSV =-=(16)-=-. Nevertheless,sreinfections are very common throughout life. In older children and adults, they are usually associatedswith milder disease, indicating that RSV infections induce only partial immunity...

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...ing the second year have been observed (16). The incidence of RSV-associatedsbronchiolitis and pneumonia was highest in 2- to 3-month-old infants, whereas with increasing age, thesincidence decreased =-=(36, 47)-=-. In this study, RSV was detected in over 1,400 respiratory samplessover a period of nine consecutive seasons. Most infected individuals were babies 0 to 3 months oldsaccounting for 51% of all samples...

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...hescarbohydrates of the G protein are O glycosylated. Thereby serine and threonine residues arespotential attachment sites for O-linked sugars, and the G protein has an unusually high content of boths=-=(9)-=-. The analysis of 89 amino acids by the program NetOglyc predicted 22 to 34 serine and threoninesresidues to be potentially O glycosylated with score predicators (G scores) of 0.5 and 0.8.sInteresting...

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...ivergence values determined weresdescribed in terms of mean and standard deviation. To estimate the numbers of potentially Oglycosylated serine and threonine residues, NetOglyc software (version 3.1) =-=(18)-=- was used.sSynonymous and nonsynonymous mutations were analyzed by the method of Nei and Gojobori (31).sThe program SNAP (Synonymous/Nonsynonymous Analysis Program) provided by the humansimmunodeficie...

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...South Africa; A5.27 and NG/009/02 from Japan; A5.16 and Sal/173/99 from Brazil;sand A7.2 and CN1973 from Canada), suggesting that RSV group A strains are distributed worldwidesas described previously =-=(5, 15)-=-.sA reason for the long-lasting circulation of genotypes GA2 and GA5 in Germany might be due tosmutations in their amino acid sequences. The nucleotide divergence determined for genotype GA2swas 3.7%,...

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...South Africa; A5.27 and NG/009/02 from Japan; A5.16 and Sal/173/99 from Brazil;sand A7.2 and CN1973 from Canada), suggesting that RSV group A strains are distributed worldwidesas described previously =-=(5, 15)-=-.sA reason for the long-lasting circulation of genotypes GA2 and GA5 in Germany might be due tosmutations in their amino acid sequences. The nucleotide divergence determined for genotype GA2swas 3.7%,...

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...h has only 44% amino acid identity between the two groups compared with 83% forsthe transmembrane and cytoplasmic domains (21). The G protein is heavily glycosylated with N-linkedsand O-linked sugars =-=(60)-=-. Mainly, the ectodomain of the G protein has a high content of serine andsthreonine residues which are potential acceptor sites for O-linked sugars. However, the amino acidssequence positions of pote...

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... occur yearly in the late fall, winter, or spring but not insthe summer (22, 29, 61). Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics =-=(2, 4, 13, 20)-=-, although group A viruses tend to predominates(8).sThere is limited information regarding the molecular epidemiology of RSV in Germany. For the firststime, we describe both the genetic variability of...

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...her European countries, such as Switzerland, Sweden, andsFinland, observed a yearly varying pattern of early and late seasons, whereas early seasons aresassociated with high incidences and vice versa =-=(10, 41, 53)-=-. Previous reports from Germany includingsregions around Kiel, Stuttgart, and Freiburg also described a biennial rhythm of severe early and weakslate seasons (3, 50, 58). An explanation for the absenc...

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...and may be associated with extensive Osglycosylation of the G protein (6).sDiscussionsRSV is the most common viral pathogen causing lower respiratory tract infections among infants andsyoung children =-=(48)-=-. Infection rates for RSV of 68.8/100 children during the first year of life ands82.6/100 during the second year have been observed (16). The incidence of RSV-associatedsbronchiolitis and pneumonia wa...

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...ssinclude the second variable region of the G protein, coding for the Cterminal end of the protein. EightsRSV group A genotypes, named GA1 to GA7 and South Africa A1 (SAA1) have been described so fars=-=(38, 39, 51)-=-. Genotype analysis of RSV group A is limited to particular countries worldwide. In Europe,sfor instance, genotypic RSV group A data are provided only from Belgium and Sweden (40, 62, 63).sVariability...

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...in the number and distribution patterns of glycosylation sites can influence thesexpression of certain epitopes, and therefore, recognition of RSV by carbohydrate-specific antibodiesscan be inhibited =-=(27, 34, 35)-=-. The predicted O-glycosylation sites in the second hypervariable region ofsthe G-protein gene analyzed in this study included 22 to 34 residues. For the same region, 8 to 10spotential O-linked glycos...

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...sites are poorly conserved,salthough the general locations of the latter are similar (21). Amino acid variations of the protein exist insboth groups, but variations are more pronounced in RSV group A =-=(39, 51)-=-. Most molecular analysessinclude the second variable region of the G protein, coding for the Cterminal end of the protein. EightsRSV group A genotypes, named GA1 to GA7 and South Africa A1 (SAA1) hav...

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... occur yearly in the late fall, winter, or spring but not insthe summer (22, 29, 61). Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics =-=(2, 4, 13, 20)-=-, although group A viruses tend to predominates(8).sThere is limited information regarding the molecular epidemiology of RSV in Germany. For the firststime, we describe both the genetic variability of...

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...ribed so fars(38, 39, 51). Genotype analysis of RSV group A is limited to particular countries worldwide. In Europe,sfor instance, genotypic RSV group A data are provided only from Belgium and Sweden =-=(40, 62, 63)-=-.sVariability between RSV strains is one of the features of RSV infections that might contribute to thesability of the virus to infect people repeatedly and cause yearly outbreaks (38). RSV has a clea...

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...y published as:sReiche, J., Schweiger, B.sGenetic variability of group A human respiratory syncytial virus strains circulating in Germanysfrom 1998 to 2007s(2009) Journal of Clinical Microbiology, 47 =-=(6)-=-, pp. 1800-1810.sDOI: 0.1128/JCM.02286-08sThis is an author manuscript.sThe definitive version is available at: http://jcm.asm.org/sGenetic Variability of Group A Human Respiratory SyncytialsVirus Str...

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...elgium, Uruguay, Argentina, andsthe United States (12, 14, 39, 40, 62). In these and other countries, mainly the genotypes GA2 andsGA5 and in a few cases genotype GA7 have been circulating since 1998 =-=(12, 32, 37, 44)-=-. Thescontinuous and predominant circulation of genotypes GA2 and GA5 demonstrate that these genotypessare stable and have become epidemic in many countries.sViruses isolated in Germany comprise sets ...

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...in the number and distribution patterns of glycosylation sites can influence thesexpression of certain epitopes, and therefore, recognition of RSV by carbohydrate-specific antibodiesscan be inhibited =-=(27, 34, 35)-=-. The predicted O-glycosylation sites in the second hypervariable region ofsthe G-protein gene analyzed in this study included 22 to 34 residues. For the same region, 8 to 10spotential O-linked glycos...

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... therefore, reviewed, andsclosely related strains, based on nucleotide similarity values of ≥96.6%, were further divided intosgroups that we arbitrarily designated subtypes (according to Peret et al. =-=[39]-=-). Genotypes GA2 andsGA5 showed a high variability with 20 and 28 subtypes, respectively. GA7 includes only two subtypess(Fig. 2).sSeasonal circulation pattern of RSV group A genotypes and subtypes.sP...

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... Nevertheless,sreinfections are very common throughout life. In older children and adults, they are usually associatedswith milder disease, indicating that RSV infections induce only partial immunity =-=(19)-=-.sRSV strains are separated into two major groups based on antigenic and genetic variability. The mainsdifferences between RSV groups A and B were found in the attachment glycoprotein G (1, 6, 30).sVa...

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...ribed so fars(38, 39, 51). Genotype analysis of RSV group A is limited to particular countries worldwide. In Europe,sfor instance, genotypic RSV group A data are provided only from Belgium and Sweden =-=(40, 62, 63)-=-.sVariability between RSV strains is one of the features of RSV infections that might contribute to thesability of the virus to infect people repeatedly and cause yearly outbreaks (38). RSV has a clea...

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...are usually low (7, 26, 55). Significant boosting, particularly for the immunoglobulin Gsand immunoglobulin A responses, will develop soonest when children are reinfected for the second orsthird time =-=(43)-=-.sRSV was isolated from autumn to spring in all nine seasons investigated herein. Thus, RSV isscirculating mainly in the winter months as determined by other studies of the temperate zone (23).sMonito...

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...igh incidences and vice versa (10, 41, 53). Previous reports from Germany includingsregions around Kiel, Stuttgart, and Freiburg also described a biennial rhythm of severe early and weakslate seasons =-=(3, 50, 58)-=-. An explanation for the absence of the biennial pattern in our study might besdue to missed cases during summer months which would influence the onset of a season. Apart fromsthat, we observed no fre...

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...nfected during the first months of life. Both serum and secretorysantibodies are produced in response to infection, even by very young infants; in this group, however,stiters achieved are usually low =-=(7, 26, 55)-=-. Significant boosting, particularly for the immunoglobulin Gsand immunoglobulin A responses, will develop soonest when children are reinfected for the second orsthird time (43).sRSV was isolated from...

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... occur yearly in the late fall, winter, or spring but not insthe summer (22, 29, 61). Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics =-=(2, 4, 13, 20)-=-, although group A viruses tend to predominates(8).sThere is limited information regarding the molecular epidemiology of RSV in Germany. For the firststime, we describe both the genetic variability of...

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...sing age, the RSV prevalence decreased toslevels of 23% in children between 1 and 3 years old. Long-term studies from Germany analyzing thesimpact of RSV in hospitalized children confirm our findings =-=(3, 57)-=-. These data revealed that 22.8% tos42.8% were positive for RSV in the age group between 0 and ≤ 12 months (54, 56). In children oldersthan 1 year, the prevalence of RSV was decreasing (54). The frequ...

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...rom Germany analyzing thesimpact of RSV in hospitalized children confirm our findings (3, 57). These data revealed that 22.8% tos42.8% were positive for RSV in the age group between 0 and ≤ 12 months =-=(54, 56)-=-. In children oldersthan 1 year, the prevalence of RSV was decreasing (54). The frequency of RSV particularly in smallschildren might be caused by their immune status. Although high levels of maternal...

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...bserved in Belgium and Argentina where RSV group A viruses predominated for two consecutivesseasons (52, 63). A study in India reported on RSV group A dominance for three consecutive epidemicsseasons =-=(37)-=-. RSV group B predominated for a single season in the countries investigated and at thessame or similar time as observed in Germany. Thus, it can be concluded that RSV group Aspredominated within simi...

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...in the number and distribution patterns of glycosylation sites can influence thesexpression of certain epitopes, and therefore, recognition of RSV by carbohydrate-specific antibodiesscan be inhibited =-=(27, 34, 35)-=-. The predicted O-glycosylation sites in the second hypervariable region ofsthe G-protein gene analyzed in this study included 22 to 34 residues. For the same region, 8 to 10spotential O-linked glycos...

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...virus to infect people repeatedly and cause yearly outbreaks (38). RSV has a clearsseasonality. In temperate climates, outbreaks occur yearly in the late fall, winter, or spring but not insthe summer =-=(22, 29, 61)-=-. Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics (2, 4, 13, 20), although group A viruses tend to predominates(8).sThere is limited i...

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...her European countries, such as Switzerland, Sweden, andsFinland, observed a yearly varying pattern of early and late seasons, whereas early seasons aresassociated with high incidences and vice versa =-=(10, 41, 53)-=-. Previous reports from Germany includingsregions around Kiel, Stuttgart, and Freiburg also described a biennial rhythm of severe early and weakslate seasons (3, 50, 58). An explanation for the absenc...

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...ing the second year have been observed (16). The incidence of RSV-associatedsbronchiolitis and pneumonia was highest in 2- to 3-month-old infants, whereas with increasing age, thesincidence decreased =-=(36, 47)-=-. In this study, RSV was detected in over 1,400 respiratory samplessover a period of nine consecutive seasons. Most infected individuals were babies 0 to 3 months oldsaccounting for 51% of all samples...

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...d insseveral other countries (Table 8). A regular three-year cyclic pattern of group dominance wassobserved in Belgium and Argentina where RSV group A viruses predominated for two consecutivesseasons =-=(52, 63)-=-. A study in India reported on RSV group A dominance for three consecutive epidemicsseasons (37). RSV group B predominated for a single season in the countries investigated and at thessame or similar ...

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...nfected during the first months of life. Both serum and secretorysantibodies are produced in response to infection, even by very young infants; in this group, however,stiters achieved are usually low =-=(7, 26, 55)-=-. Significant boosting, particularly for the immunoglobulin Gsand immunoglobulin A responses, will develop soonest when children are reinfected for the second orsthird time (43).sRSV was isolated from...

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...her European countries, such as Switzerland, Sweden, andsFinland, observed a yearly varying pattern of early and late seasons, whereas early seasons aresassociated with high incidences and vice versa =-=(10, 41, 53)-=-. Previous reports from Germany includingsregions around Kiel, Stuttgart, and Freiburg also described a biennial rhythm of severe early and weakslate seasons (3, 50, 58). An explanation for the absenc...

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...her geographic areas. Between 1964 and 1996, genotypes GA1, GA3, and GA4 circulatedssporadically in addition to the genotypes GA2 and GA5 in Sweden, Belgium, Uruguay, Argentina, andsthe United States =-=(12, 14, 39, 40, 62)-=-. In these and other countries, mainly the genotypes GA2 andsGA5 and in a few cases genotype GA7 have been circulating since 1998 (12, 32, 37, 44). Thescontinuous and predominant circulation of genoty...

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...nfected during the first months of life. Both serum and secretorysantibodies are produced in response to infection, even by very young infants; in this group, however,stiters achieved are usually low =-=(7, 26, 55)-=-. Significant boosting, particularly for the immunoglobulin Gsand immunoglobulin A responses, will develop soonest when children are reinfected for the second orsthird time (43).sRSV was isolated from...

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...epidemic seasons.sWorldwide, the respiratory syncytial virus (RSV) is the major pathogen of lower respiratory tractsinfections in infants and young children in both developing and developed countries =-=(46)-=-. By the age ofs2 years, virtually all children have been infected at least once with RSV (16). Nevertheless,sreinfections are very common throughout life. In older children and adults, they are usual...

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... occur yearly in the late fall, winter, or spring but not insthe summer (22, 29, 61). Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics =-=(2, 4, 13, 20)-=-, although group A viruses tend to predominates(8).sThere is limited information regarding the molecular epidemiology of RSV in Germany. For the firststime, we describe both the genetic variability of...

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...elgium, Uruguay, Argentina, andsthe United States (12, 14, 39, 40, 62). In these and other countries, mainly the genotypes GA2 andsGA5 and in a few cases genotype GA7 have been circulating since 1998 =-=(12, 32, 37, 44)-=-. Thescontinuous and predominant circulation of genotypes GA2 and GA5 demonstrate that these genotypessare stable and have become epidemic in many countries.sViruses isolated in Germany comprise sets ...

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...ribed so fars(38, 39, 51). Genotype analysis of RSV group A is limited to particular countries worldwide. In Europe,sfor instance, genotypic RSV group A data are provided only from Belgium and Sweden =-=(40, 62, 63)-=-.sVariability between RSV strains is one of the features of RSV infections that might contribute to thesability of the virus to infect people repeatedly and cause yearly outbreaks (38). RSV has a clea...

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...igh incidences and vice versa (10, 41, 53). Previous reports from Germany includingsregions around Kiel, Stuttgart, and Freiburg also described a biennial rhythm of severe early and weakslate seasons =-=(3, 50, 58)-=-. An explanation for the absence of the biennial pattern in our study might besdue to missed cases during summer months which would influence the onset of a season. Apart fromsthat, we observed no fre...

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...the summer (22, 29, 61). Both RSV groups can be present in the same community, and their relativesproportions may differ between epidemics (2, 4, 13, 20), although group A viruses tend to predominates=-=(8)-=-.sThere is limited information regarding the molecular epidemiology of RSV in Germany. For the firststime, we describe both the genetic variability of RSV group A viruses and the circulation pattern o...

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...rom Germany analyzing thesimpact of RSV in hospitalized children confirm our findings (3, 57). These data revealed that 22.8% tos42.8% were positive for RSV in the age group between 0 and ≤ 12 months =-=(54, 56)-=-. In children oldersthan 1 year, the prevalence of RSV was decreasing (54). The frequency of RSV particularly in smallschildren might be caused by their immune status. Although high levels of maternal...

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...sing age, the RSV prevalence decreased toslevels of 23% in children between 1 and 3 years old. Long-term studies from Germany analyzing thesimpact of RSV in hospitalized children confirm our findings =-=(3, 57)-=-. These data revealed that 22.8% tos42.8% were positive for RSV in the age group between 0 and ≤ 12 months (54, 56). In children oldersthan 1 year, the prevalence of RSV was decreasing (54). The frequ...

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...her geographic areas. Between 1964 and 1996, genotypes GA1, GA3, and GA4 circulatedssporadically in addition to the genotypes GA2 and GA5 in Sweden, Belgium, Uruguay, Argentina, andsthe United States =-=(12, 14, 39, 40, 62)-=-. In these and other countries, mainly the genotypes GA2 andsGA5 and in a few cases genotype GA7 have been circulating since 1998 (12, 32, 37, 44). Thescontinuous and predominant circulation of genoty...