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Antigenic and genetic characterization of serotype G2 human rotavirus strains from South Africa from 1984 to 1998.

Within South Africa, cyclic peaks of serotype G2P[4] rotavirus infection have been observed and these strains were prevalent in some locations. To examine the cyclic phenomenon of serotype G2 rotaviruses, historical stool collections from South Africa spanning 15 years were screened for G2 strains. Subgroup (VP6) ELISA, polyacrylamide gel electrophoresis (PAGE), and P genotyping were performed on 43 G2 strains to investigate the associated DS-1 genogroup characteristics. Antigenic variation of the gene encoding the major neutralization glycoprotein (VP7) was also investigated using G2-specific monoclonal antibodies. In addition, the VP7 gene of 14 serotype G2 strains was sequenced to examine genetic variation. Serotype G2 strains from South Africa displayed a 10 year cyclic pattern with major epidemics occurring in 1987 and 1997. Serotype G2 strains were also found co-dominant with G(1) strains in 1984, 1990, and 1993. The G2 strains from the major epidemics appeared to have emerged from community strains in a manner similar to that suggested for G(1) strains The serotype G2 strains displayed subgroup I specificity and short electropherotypes characteristic of DS-1 genogroup rotavirus strains but appeared to differ in the VP4 gene. Genetic analyses revealed three major serotype G2 lineages, i.e., strains isolated prior to 1987, strains isolated between 1988 and 1994, and strains isolated from 1995. The use of monoclonal antibodies and PCR primers designed against older G2 strains has resulted in the failure to serotype G2 strains circulating currently.

Amino Acid Sequence↗

Electropherotypes and serotypes of human rotavirus in Estonia in 1989-1992.

The electropherotypes and serotypes of human rotaviruses circulating in Tallinn (Estonia) in 1989-1992 have been studied. Rotaviruses were found in 372 (25.8%) of 1,442 faecal specimens of pediatric patients with acute diarrhea. Polyacrylamide gel electrophoresis of RNA from 318 isolates has revealed 41 electropherotypes. Frequent genomic alterations, including a shift of predominant electropherotypes, were observed during the study period. The serotype of 158 (50.3%) of 314 tested rotavirus isolates was identified using ELISA with VP7-specific monoclonal antibodies against 4 established human rotavirus serotypes. Serotype G1 was found to be largely predominant and accounted for 70.9% of the typeable specimens, serotypes G4, G2, and G3 accounted for 12%, 9.5%, 7.6%, respectively. A major shift to serotype G4 took place in 1990-1991. Serotype G1 was represented by the largest number of electropherotypes. All G1, G3 and G4 isolates were of "long" and all G2 isolates were of "short" electropherotypes. According to our results the isolates of an identical electropherotype belong to the same serotype.

Antibodies, Monoclonal↗

Comparative amino acid sequence analysis of the major outer capsid protein (VP7) of porcine rotaviruses with G3 and G5 serotype specificities isolated in Venezuela and Argentina.

Seven porcine group A rotavirus strains isolated in Venezuela were shown to be antigenically related to serotype G3 (five strains) or to serotype G5 (two strains), whereas two strains isolated in Argentina were classified as serotype G5. The serological classification of eight of these strains was confirmed by sequence analysis of the gene encoding the VP7 glycoprotein. A high degree of homology was observed among strains belonging to the same G serotype, although some variations in the serotype-specific regions were detected among different strains. Comparison with the published VP7 amino acid sequences of serotype G3 indicated that most porcine rotavirus strains are more closely related to each other and to human rotavirus strains than to rotavirus strains isolated from other species. Amino acid sequence comparison among serotype G5 porcine strains revealed that Venezuelan porcine isolates were more closely related to the American strain OSU, while the Argentinian strains had a higher similarity to the Australian strain TRF-41. This report confirms the worldwide distribution of these G serotypes among the porcine population.

Amino Acid Sequence↗

Molecular epidemiology of hepatitis B viral serotypes and genotypes in taiwan.

Subtypes of hepatitis B virus (HBV) have specific geographic distributions and can serve as epidemiological markers. The relationship of HBV serotypes and genotypes in Taiwan and their correlation with the domiciles of origin in 122 patients with chronic HBV infection were investigated. The serotype of HBV was determined by comparing the surface gene encoding amino acids 22-148 of the major surface protein with published sequences. Genotyping of HBV was performed by polymerase chain reaction-restriction fragment length polymorphism. Serotype adw accounted for 70% (85/122) of all HBVs, with the remaining belonging to serotype adr. All adr HBVs were genotype C, regardless of the patient's domicile. Of the 85 adw HBVs, 69 (81%) were genotype B, 10 (12%) were genotype C, 5 (6%) were genotype F and only 1 (1%) was genotype A. In the 31 patients originating from mainland China, the prevalence of adr/genotype C was higher than in the 91 Taiwanese patients (15/31 vs. 22/91; p < 0.05). The distribution of the HBV serotypes and genotypes was not significantly different between 17 patients born in Taiwan (6 adw/genotype B, 2 adw/genotype C, 1 adw/genotype F and 8 adr/genotype C) and 14 patients born in mainland China (5 adw/genotype B, 2 adw/genotype C and 7 adr/genotype C). Our results indicate that in Taiwan, most HBVs of serotype adw are genotype B, and all HBVs of serotype adr are genotype C. Patients with origins in mainland China have a higher proportion of serotype adr/genotype C infection.

Adult↗

Coexistence of different serotypes of dengue virus.

We formulate a non-linear system of differential equations that models the dynamics of dengue fever. This disease is produced by any of the four serotypes of dengue arbovirus. Each serotype produces permanent immunity to it, but only a certain degree of cross-immunity to heterologous serotypes. In our model we consider the relation between two serotypes. Our interest is to analyze the factors that allow the invasion and persistence of different serotypes in the human population. Analysis of the model reveals the existence of four equilibrium points, which belong to the region of biological interest. One of the equilibrium points corresponds to the disease-free state, the other three equilibria correspond to the two states where just one serotype is present, and the state where both serotypes coexist, respectively. We discuss conditions for the asymptotic stability of equilibria, supported by analytical and numerical methods. We find that coexistence of both serotypes is possible for a large range of parameters.

Aedes↗

Serotyping and genotyping of hepatitis C virus in Belgium.

BACKGROUND: Given that both pathogenicity and the response to treatment are possibly associated with hepatitis C virus (HCV) serotype, it appeared sensible to establish the prevalence of the different HCV types in Belgium. MATERIALS AND METHODS: The HCV serotypes were determined in 68 HCV-RNA and anti-HCV-positive samples taken from Belgian patients and compared with the results of the genotyping assay. Possible associations with age and sex were investigated. RESULTS: Antibodies were identified in 55 (80.9%) of the 68 samples, with serotype 1 (58.8%) and serotype 3 (19.1%) showing the highest prevalence. 17 samples contained several serotypes with serotype 1 being detected in 82.4% of cases. Nine of the 11 samples undetermined by serotyping could be determined by genotyping. There was no significant difference in the distribution of HCV types with respect to gender. Compared with genotype 3 (p < 0.01) and genotypes 2 and 4 (p = 0.05), genotype 1 was detected among older patients. CONCLUSION: Our data showed a 96.0% correlation between the serotyping and genotyping assays. Genotypes 1 and 3 are the most prevalent types among Belgian patients. The data suggest that genotype 1 spread earlier than genotypes 2, 3 and 4. This corroborates previous European studies.

Adult↗

Monoclonal antibody analysis of serotype-restricted and unrestricted bluetongue viral antigenic determinants.

Twenty-one monoclonal antibodies that react with bluetongue virus (BTV) and have restricted or unrestricted serotype specificities were identified in culture supernatants of hybridomas derived from lymphocytes of mice immunized with BTV serotype 17. Hybridomas were screened and antibody specificities characterized in a solid-phase radioimmunoassay and by immunoprecipitation with radiolabeled, BTV 17-infected cell lysates. Three general serotype specificities were demonstrated by 13 antibodies that precipitated structural viral protein 9 (VP 9). One antibody precipitated VP 7, a 48,000 dalton nonstructural protein, and reacted in radioimmunoassay with 20 BTV serotypes and Epizootic Hemorrhagic Disease of Deer Virus serotype 1 (EHDV 1), EHDV 2, and Ibaraki virus, but not with uninfected cells. One serotype-specific antibody neutralized infectivity, inhibited hemagglutination by BTV 17, and precipitated VP 2 and VP 3. A second antibody, with restricted serotype specificity, precipitated VP 2 and VP 8. These results confirm those of others (Huismans and Erasmus, Onderstepoort J. Vet. Res. 48, 51-58, 1981) that BTV serotype-specific and neutralizing epitopes are associated with VP 2.

Antibodies, Monoclonal↗

The major and minor group receptor families contain all but one human rhinovirus serotype.

Previous studies have assigned 88 human rhinovirus (HRV) serotypes to major and minor receptor groups. Extension of these studies to include the remaining 14 unassigned serotypes indicated that 13 serotypes belong to the major group since their infection of HeLa cells is completely blocked by a monoclonal antibody that recognizes the major group receptor. This result indicates that the major group now accounts for 91 of the 102 known serotypes, while the minor group contains 10 serotypes. One serotype, HRV-87, appears to utilize neither the major nor minor group receptor and may represent a third receptor group. HRV-87 attachment cannot be blocked by other serotypes and displays a binding tropism similar to but distinct from minor group viruses. Unlike major and minor group serotypes, HRV-87 attachment and infection requires the presence of sialic acid on cellular receptors.

Animals↗

Serotypes of Escherichia coli strains isolated from piglets with diarrhea in swine industrial farms.

Serotypes of E. coli strains isolated from piglets, which died with symptoms of diarrhea in 9 swine industrial farms, were determined. Large numbers of serotypes (from 16 to 27) in individual farms were detected. The sets of serotypes from 9 investigated farms differed among each other significantly, depending on the farm and time of examination. It was found that more than one serotype of E. coli may exist in the pig body and contribute to the development of disease. The predominant serotypes, i.e. those comprising more than 10% of serologically determined strains, were found to exist in 6 of the investigated farms and not in the remaining ones. Among the predominant serotypes, particularly important seem to be strains with K88 antigen. For prophylaxis of piglet colibacteriosis in industrial farms in Poland two vaccines for sows are recommended: one containing the K88 antigen only and the other the following serotypes: 0149:K91,K88; 020:K57; 020:K83; O157:K88; 01:K1; 0136:K78; 024:K?; 078:K80 and 0118:K? Strains belonging to these serotypes were the most prevalent in our strain collection.

Animals↗

Serotype identification of Australian bluetongue viruses using a rapid fluorescence inhibition test.

Rapid serotyping of bluetongue virus (BTV) isolates is required to facilitate the choice of an appropriate serotype-specific vaccine in a disease situation or to improve surveillance of BTV serotype prevalence. This communication describes the development and validation of a bluetongue virus fluorescent inhibition test (BTV FIT) as a rapid method to serotype Australian BTV isolates. The BTV FIT uses virus neutralisation principles similar to those used in the rabies rapid fluorescent focus inhibition test. The BTV FIT has the ability to provide an accurate serotype identification within 24 h thereby abbreviating the serotyping process by 3-4 days relative to conventional virus neutralisation assays and making the BTV FIT comparable time-wise with the polymerase chain reaction technique. The development of the BTV FIT is described using BTV reference viruses which have been isolated in Australia, and validation of the assay by assessment of five Australian BTV isolates of unknown serotype by comparison with the plaque inhibition method. The use of the BTV FIT readily facilitated rapid and accurate serotype identification of Australian BTV reference viruses and five unknown BTV isolates with results indicating full agreement with the plaque inhibition method.

Animals↗

Phylogenetic comparison of the serotype-specific VP2 protein of bluetongue and related orbiviruses.

Regions of the VP2 gene from various bluetongue virus serotypes were sequenced and phylogenetic comparisons were performed. The sequences were characteristic for each BTV serotype and isolates of the same serotype could be grouped geographically, mimicking the topotyping characteristics of BTV VP3 gene sequences. PCR amplification and sequence analysis were used to show the close relationship between Caribbean BTV isolates and South African BTV isolates of the same serotype. Similarly, Australian BTV isolates showed a close genetic relationship with Asian BTV isolates of the same serotype. A multiple amino acid sequence alignment of fifteen BTV serotypes and other orbiviruses over a proposed major neutralization site showed this region (317 335 aa.) was highly variable and nucleotide sequences showed that BTV serotypes could be grouped into nucleotypes, or related serotypes, in broad agreement with the inter-relationships postulated by Erasmus (1990), using plaque-reduction tests.

Amino Acid Sequence↗

Immunity in pigeons against homologous and heterologous serotypes of Streptococcus bovis after infection.

Groups of 20, 20 and 10 pigeons were intravenously inoculated with 1 x 10(9) CFU of a Streptococcus bovis serotype 1, 2 and 3 strain, respectively. Groups which received the highly virulent serotypes 1 or 2 strains were treated with antibiotics starting from 2 days post inoculation in order to prevent the development of clinical signs. Pigeons inoculated with the low virulence S. bovis serotype 3 strain were left untreated. Another group of 10 non-inoculated pigeons was used as challenge control. Four weeks later, pigeons were challenged intravenously with S. bovis serotype 1. Morbidity after challenge infection was 19%, 36% and 70% in groups previously inoculated with serotypes 1, 2 and 3, respectively, whereas it was 100% in the control group. Results demonstrate that pigeons developed significant protective immunity to S. bovis serotype 1 septicaemia following infection with S. bovis serotype 1 and 2 strains but not after infection with the serotype 3 strain. Protection did not correlate with the presence of antibodies as detected in ELISA.

Animals↗

Serotypes of Streptococcus pneumoniae causing meningitis in southern India. Use of new direct latex agglutination antigen detection tests in cerebrospinal fluid.

The utility of a new latex agglutination (LA) test to directly determine serotypes of Streptococcus pneumoniae in cerebrospinal fluid (CSF) was assessed in prospective evaluation at a referral hospital in southern India. Samples from 18 ill patients with Gram-positive organisms in CSF were tested. The presence of C polysaccharide or specific serotype antigen (types 1, 3, 4, 5, 6, 12, 14, 18, 19, and 23) of S. pneumoniae was detected by slide LA test. Pneumococcal antigen was detected in 17 (94%) of 18 CSF specimens; in 14 (78%) the serotype was determined directly. Serotypes 1, 5, 6, 19, 23, 7, 10, 34, and 38 were found in these patients. A quellung test of the cultured isolates confirmed the serotypes. Information regarding the serotype distribution of pneumococci in varied geographic locations is important for the design and evaluation of pneumococcal vaccines. The slide LA tests seemed useful in detecting S. pneumoniae antigens and in determining the serotype, and have promise in simplifying the gathering of serotype data.

Adult↗

Sequential rotavirus infections: characterization of serotypes and electrophoretypes.

Few studies have characterized the rotaviruses observed in repeated infections in the same patient. Rotavirus serotypes and electrophoretypes were determined from faecal samples of 24 children hospitalized between 1984 and 1989 in a Clermont-Ferrand hospital and who had had repeated rotavirus shedding. The patients were aged between 3 days and 4 years. Fifty infections were recorded. Of the 48 serotypes obtained, 18 were serotype 1 (37.5%), 10 serotype 2 (20.8%) and 20 serotype 4 (41.7%). Serotype 3 was not found. Seven serotype 2 infections were characterized during rehospitalization. Six patients had two subsequent infections with the same serotype.

Child, Preschool↗

Preliminary study of certain serotypes, genetic and antimicrobial resistance profiles of verotoxigenic Escherichia coli (VTEC) isolated in Bosnia and Germany from cattle or pigs and their products.

The aim of this study was to gather more information on the spread of VTEC serotypes, genetic profiles and resistance patterns from pigs or pork and from cattle or beef in different areas, and to improve detection of the source of outbreaks with a wider data pool. Of 130 Escherichia coli samples isolated from a cattle slaughter house and beef retail products in Sarajevo, Bosnia and Herzegovina (BiH), seven were identified as verotoxigenic (VTEC). In comparison, 22 VTEC of 264 E. coli isolates were isolated from bovine faeces (14) and beef products (8) from Germany. Furthermore 23 VTEC of 76 isolates were identified from pig carcasses (10), faeces (9) and pork products (4) from Germany. Gene detection and serotyping were carried out in our laboratory and in the National Reference Laboratory. Antimicrobial resistance was tested with the dilution method in microtitre plates. All porcine isolates belonged to serotypes thus far not associated with human disease. Bovine VTEC were either serotypes commonly associated with human diseases (O157:H7, O103:H2, O157:H-) or rare serotypes. One serotype (O96:H19) was found only in isolates from Sarajevo. Most German VTEC, especially those of porcine origin, had only vtx2 genes, whereas all Bosnian isolates had vtx1 and vtx2 genes. The eae gene was found only in "classical" VTEC serotypes. All 52 VTEC (100%) investigated were resistant to the three sulfonamides tested; porcine isolates were mainly resistant to oxytetracycline (43%) and chlortetracycline (37%), bovine isolates mainly to trimethoprim/sulfamethoxazole and ampicillin (10% each). If sulfonamide resistances are disregarded, more than half (53.8%) of porcine VTEC were multiresistant and one-fourth (25%) of German bovine isolates, but none of the Bosnian bovine isolates. The results show the considerable spread of resistances in VTEC. These results also point out the necessity of gathering data from different geographical areas in order to be able to identify typical local variations in serotypes or gene expression and thus to trace human infections more quickly to their source.

Animals↗

Identification of the Actinobacillus pleuropneumoniae serotype using PCR based-apx genes.

Actinobacillus pleuropneumoniae (A. pleuropneumoniae) is an important swine respiratory disease pathogen, which has at least 15 serotypes. There are several techniques for the serotyping of A. pleuropneumoniae, however, these techniques are time consuming. In this study, the polymerase chain reaction (PCR) technique was developed for serotyping A. pleuropneumoniae using a set of specific primer designated for the apxI, apxII, apxIII and apxIVA genes. By this PCR typing system, 10 out of the 13 reference strains of A. pleuropneumoniae were differentiated. However, it was not possible to distinguish serotype 2 from 8, serotype 5a from 5b and serotype 9 from 11. Each serotype of A. pleuropneumoniae showed its own products patterns. The PCR typing system was further applied for typing the field isolates of A. pleuropneumoniae and compared to that using the gel immunodiffusion (GID) technique. The results from both PCR and GID techniques were in accordance. Thus, the PCR typing system may provide a rapid and useful tool for typing the serotypes of A. pleuropneumoniae.

Actinobacillus Infections↗

Passive immune cross-protection in mice produced by rabbit antisera against different serotypes of Pasteurella multocida.

Haemorrhagic septicaemia (HS) and fowl cholera (FC) are specific diseases caused by certain serotypes of Pasteurella multocida. Strains that usually cause HS in cattle and water buffalo do not produce FC in avian species, and strains that cause FC do not produce HS in cattle and water buffalo. A variety of P. multocida serotypes, including unusual strains which can cause HS in wild ruminants, were evaluated in passive immune protection studies to determine the immunological relationship between strains associated with HS and FC. Various degrees of cross-protection were seen among the strains. Antiserum against a serotype B:3,4 strain protected against strains capable of causing HS (serotypes B:1, B:2, B:3,4, B:4 and E:2) and FC (serotypes A:1, A:3 and A:5). Antiserum against an FC strain (serotype A:5) similarly protected against strains capable of causing HS and FC. Antigenic analyses indicated that cross-protection was not necessarily induced by serotype-specific capsular (beta) or somatic (gamma) antigens or major porin protein. SDS-PAGE and immunoblots of whole cell lysates of the different HS and FC strains showed many protein-staining bands with similar mobilities and antigenic activity. These cross-reactive antigenic bands occurred in the 20- to 120-kDa range. Adsorption of antiserum with a heterologous serotype removed its reactivity with most of these bands, as well as its ability to cross-protect.

Animals↗

Blocking enzyme-linked immunosorbent assay for detection of antibodies against Actinobacillus pleuropneumoniae serotype 6 in pig serum.

A blocking enzyme-linked immunosorbent assay (ELISA) detecting antibodies against Actinobacillus pleuropneumoniae (Ap) serotype 6 was developed. The blocking ELISA was based on the inhibition of a polyclonal antibody raised against Ap serotype 6. Purified lipopolysaccharide from Ap serotype 6 was used as antigen. The blocking ELISA was tested against sera from pigs experimentally infected with the 12 serotypes of Ap biotype 1. Cross-reaction with serotypes 3 and 8 but not with other serotypes was observed. The sensitivity and specificity of the test on a herd level were evaluated with sera from herds naturally infected with serotypes 2, 6, 8 or 12 and with sera from herds free of infection with any Ap serotype. The blocking ELISA showed a high herd sensitivity (1.00 (0.79-1.00)) and specificity (0.97 (0.93-0.99)).

Actinobacillus Infections↗