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Serotyping of Pasteurella multocida isolated from swine lungs collected at slaughter.

We serotyped 222 Pasteurella multocida strains isolated from swine lungs at slaughter. Capsular serotypes A and D were determined by the hyaluronidase sensitivity and acriflavin agglutination tests, respectively. Somatic antigens were determined by gel diffusion against standard antisera. Capsular serotype A was found in 97.3% of the strains and serotype D in the remaining 2.7%. The primary somatic antigen most commonly found was type 3 (86.0%). Type 5 was also very common (88.7%), but was usually a secondary antigen. The most common overall serotype was A:3(5) (39.2%). Other common serotypes were: A:3(4,5,12) (12.2%); A:3(4,5) (11.2%); A:3(5,12) (10.4%); A:3 (6.8%); and A:5 (6.8%). Type D strains had a similar distribution of serotypes, but with a higher prevalence of D:5 (33.3%) and D:3(5) (33.3%).

Animals↗

Investigation of a waterborne outbreak of Campylobacter jejuni enteritis with a serotyping scheme based on thermostable antigens.

Serotyping of 11 human and 2 water isolates of Campylobacter jejuni associated with a waterborne outbreak revealed two serotypes among the human isolates. One of these (serotype 58) was a new serotype and was added to the serotyping scheme. Serotypes were defined by using extracted thermostable antigens and passive hemagglutination titrations of both unabsorbed and cross-absorbed antisera. Two water isolates of the same serotype as six human isolates provided evidence to link a contaminated water supply to the outbreak.

Antigens, Bacterial↗

New serotypes of Treponema hyodysenteriae.

Three isolates of pathogenic Treponema hyodysenteriae, B8044, B6933, and Ack 300/8, were serotyped by double immunodiffusion using lipopolysaccharide antigen and rabbit antiserum. The precipitation reactions of these three isolates were then compared with precipitin reactions from the reference strains of serotypes 1 through 4. The data indicated that B8044, B6933, and Ack 300/8 share antigenic determinants with either serotype 1 or 2 or both. However, when the cross-reacting antibodies were removed by adsorption, all reactions were found to be monospecific. Based on these results, three new serotypes of pathogenic T. hyodysenteriae are proposed: B8044 serotype 5, B6933 serotype 6, and Ack 300/8 serotype 7.

Animals↗

Occurrence and distribution of serotypes of the Arizona subgroup of Salmonella strains in the United States from 1967 to 1976.

The Salmonella Arizona subgroup contains gram-negative enteric bacteria that are closely related to other salmonellae biochemically, serologically, and genetically. Although the Arizona subgroup may be isolated from a wide variety of nonhuman and human sources, the arizonae are uncommonly recognized as human pathogens, and surprisingly little is known about their epidemiology. From 1967 through 1976, the Centers for Disease Control received 858 Arizona subgroup cultures from human and nonhuman sources representing 143 different serotypes in 33 somatic groups; several serotypes had not been previously reported. The 374 cultures from humans represent 71 different serotypes; extraintestinal isolates were present in 31 (44%) serotypes. Compared with data from a previous 20 years of surveillance, the proportion of Arizona subgroup strains isolated from stools, blood, and other sites was remarkably stable, but several serotypes showed marked changes in their frequency of isolation. In total, the ratio of extraintestinal to intestinal isolates was 0.37, but marked serotype-specific variation was noted, suggesting differences in virulence associated with serotype.

Amphibians↗

Rotavirus epidemiology in Vellore, south India: group, subgroup, serotype, and electrophoretype.

Rotaviruses were detected in 163 of 916 (17.8%) specimens collected from children under 3 years of age with gastroenteritis in Vellore, South India, between August 1983 and July 1985. Rotaviruses were detected throughout the study period, with a peak prevalence in December to February (winter) and June to August (southwest monsoon season). A total of 117 rotavirus strains were tested for subgroup, serotype, and rotavirus double-stranded RNA electrophoretic migration pattern; 24.8% of the strains were subgroup I, 69.2% were subgroup II, and 6.0% were neither subgroup I nor subgroup II. Subgroup I and II strains were circulating concurrently throughout the study. Of the 117 rotavirus strains, 32 (27.4%) were serotyped; 15 were serotype 1, 3 were serotype 2, 2 were serotype 3, and 12 were serotype 4. Three serotypes were circulating concurrently during the periods of peak rotavirus prevalence. In 100 of the 117 strains (85.4%) an RNA pattern was detected. One unusual subgroup I group A rotavirus with a long migration pattern and four atypical rotaviruses serologically related to group C were also detected.

Child↗

Subdivision of O serotypes of Pseudomonas aeruginosa with monoclonal antibodies.

Sixteen murine monoclonal antibodies (MAbs) against serotypes O2, O5, and O16 (serogroup II) and subtypes O2b and O5d of Pseudomonas aeruginosa were evaluated by agglutination and enzyme-linked immunosorbent assay. Six MAbs that exhibited different specificities were compared with absorbed rabbit O-type antisera for the serotyping of 55 clinical isolates of serogroup II. There was good agreement between the antibodies for strains of serotypes O2 and O2b, but MAbs revealed reproducible differences between strains that were indistinguishable with rabbit antisera. The greater serotype specificity of MAbs was illustrated by the fact that only 5 of 20 strains which were agglutinated by rabbit antiserum O16 reacted with the MAb to that serotype. One antibody, M89, that reacted with 27 of 55 serogroup II strains, apparently bound to core lipopolysaccharide epitopes. Three MAbs to the frequent serotype O6 identified six subtypes, one of which accounted for over half of the clinical strains, while two subtypes were represented by single strains only. Overall, MAbs provided a greater discrimination between strains of P. aeruginosa of the same serotype than did absorbed polyclonal antisera.

Agglutination Tests↗

Cross-reactions between Actinobacillus (Haemophilus) pleuropneumoniae strains of serotypes 1 and 9.

Actinobacillus pleuropneumoniae strains of serotypes 1 and 9 were studied for their serological properties by means of agglutination, coagglutination (CoA), indirect hemagglutination (IHA), Co-IHA, ring precipitation (RP), and immunodiffusion (ID) tests. Particulate and soluble antigens of unheated and heat-treated bacterial cells were used in various serological tests. Agglutination, CoA, and RP tests demonstrated common antigens between strains of serotypes 1 and 9. Quantitative estimation of serotype-specific antigenic activity by CoA, RP, and ID tests proved useful in differentiating strains of serotypes 1 and 9. IHA and Co-IHA tests using sheep erythrocytes sensitized with unheated or heat-treated whole-cell saline extract and the ID test using boiled whole-cell saline extract as antigen distinguished the strains of serotypes 1 and 9. In studies of absorption of rabbit antisera with heterologous whole-cell antigens there was no absorption of antibodies in tube agglutination and IHA tests, suggesting that serotype 1 and 9 strains belong to two distinct serogroups. It appears that the cross-reactivity between serotype 1 and 9 strains could be due to common epitopes associated with cell wall antigens.

Actinobacillus↗

Geographic distribution, frequency, and specimen source of Mycobacterium avium complex serotypes isolated from patients with acquired immunodeficiency syndrome.

Isolates of Mycobacterium avium complex from 727 patients with acquired immunodeficiency syndrome (AIDS) were submitted by medical centers across the United States to the Centers for Disease Control for serotyping. We were able to type 630 (87%) of these isolates by our seroagglutination procedure. Almost all typeable isolates were M. avium (serotypes 1 to 6 and 8 to 11). Blood was the major specimen source for both M. avium and the nontypeable isolates. M. intracellulare serotypes made up only 3% of all isolates from AIDS patients, with sputum being the major specimen source. More than 50% of the isolates originated from either New York or California, with serotype 4 being isolated most frequently in New York and serotype 8 appearing most frequently in California. AIDS patients in Los Angeles had a significantly higher isolation frequency for serotype 8 and a significantly lower one for serotype 4 in comparison with patients in either San Francisco or New York City.

Acquired Immunodeficiency Syndrome↗

Serotype 3 human rotavirus strains with subgroup I specificity.

During an epidemiological study on human rotavirus (HRV) infections in Italy, three subgroup I strains not associated with serotype 2 reactivity were detected. All three strains were serotype 3, each with a distinct RNA pattern showing fast-moving tenth and eleventh segments (long electropherotype). Following successful adaptation to growth in cell cultures, the serotype 3 strains (MZ58, PCP5, and PA710) were further characterized by neutralization and by RNA-RNA (Northern blot) hybridization. Antiserum to reference HRV strain YO (subgroup II, serotype 3), as well as a monoclonal antibody to VP7 of YO neutralized, at comparable titers, the homologous virus, the three unusual HRV strains, and two reference simian strains (SA11 and RRV-2, both subgroup I, serotype 3), whereas SA11 antiserum and a monoclonal antibody to VP7 of SA11 neutralized simian strains more efficiently. However, antiserum to PCP5 neutralized the three unusual isolates and the simian strains at significantly higher titers than it did with reference strain YO. With 32P-labeled RNA from MZ58 as a probe, a high degree of homology was detected by Northern blot hybridization with strains PCP5, PA710, SA11, and UK (bovine rotavirus) at the level of several segments and with strain YO only at the level of genes 7 to 9. Conversely, labeled RNA of strain YO hybridized extensively with Wa (subgroup II, serotype 1 HRV strain) but only at the level of genes 7 to 9 with MZ58, PCP5, PA710, SA11, and UK. Finally, the labeled SA11 probe hybridized at the level of RNA segments 1 to 3 and 6 to 11 to the three unusual strains. These findings suggest that the unusual subgroup I, serotype 3, strains isolated from humans are more likely to be animal rotaviruses rather than natural reassortants between different HRV strains.

Blotting, Northern↗

Epidemiological survey of human rotavirus serotypes and electropherotypes in young children admitted to two children's hospitals in northeast London from 1984 to 1990.

A retrospective and prospective survey was carried out to determine the relative frequency of rotavirus serotypes infecting children with diarrhea or vomiting or both who were admitted to the Hospitals for Sick Children in London during a 6-year period from 1984 to 1990. The results were compared with data for the same period from a study in Birmingham, United Kingdom. The serotype of rotaviruses infecting 1,019 children was ascertained by enzyme immunoassay with VP7-specific monoclonal antibodies. In London, serotype G1 accounted for 60% of the cases, serotype G4 accounted for 24%, serotype G2 accounted for 11%, G3 accounted for 3%, and coinfections accounted for 2%. Considerable differences in the relative prevalence of serotypes were seen when data from London and Birmingham were compared. A major shift from serotype G1 to G4 was observed in London in the 1989 to 1990 season, and a lesser shift was seen in Birmingham. Examination of the electrophoretic profiles of 611 rotaviruses from London showed that there were at least 108 different profiles. Continuous variation occurred throughout the 6-year period, and the same electropherotype never recurred once it had disappeared from the population. None of the electrophoretic profiles were characteristic of group B or group C rotaviruses. There was no evidence that any strain of rotavirus had become endemic in either of the children's hospitals in London.

Child, Preschool↗

Neutralizing antibodies to heterologous animal rotavirus serotypes 5, 6, 7, and 10 in sera from Ecuadorian children.

Serum samples from 870 Ecuadorian children who underwent natural rotavirus exposure were tested for neutralizing serum antibody to heterologous animal rotavirus (RV) serotypes. Six percent of the sera neutralized porcine RV OSU (serotype 5), 10% neutralized bovine RV NCDV (serotype 6), 4% neutralized avian RV Ch-2 (serotype 7), and 8% neutralized bovine RV V1005 (serotype 10). Neutralization was defined as a 90% reduction in infectious virus at a 1:100 serum dilution. The prevalence of antibody to all four heterotypic viruses increased with the age of the children and the number of human RV serotypes neutralized, but prevalences did not differ significantly between children from rural and urban areas of Ecuador. No serum sample that specifically neutralized bovine RV NCDV was identified. We inferred from the seroepidemiological analysis that human RVs contain immunorecessive neutralization epitopes that can stimulate cross-neutralizing antibody to heterotypic animal RVs. This occurs increasingly with age and with the number of human serotypes recognized by a child's neutralizing antibody. Thus, it appears that a broadened immune response to the heterotypic strains occurs with repetitive RV infections.

Animals↗

Evaluation of commercial antisera for serotyping heat-labile antigens of Campylobacter jejuni and Campylobacter coli.

Commercial antisera for serotyping 22 heat-labile antigens of Campylobacter jejuni and Campylobacter coli were evaluated by using 66 isolates from human and nonhuman sources. Test results were compared with results of tests using antisera produced at the Centers for Disease Control (CDC), Atlanta, Ga. All strains (three isolates of each of the 22 serotypes) were typeable with the CDC antisera. Of 66 test strains, 39 (59%) were typed as the same serotype with both sets of antisera. Twenty-four strains (36%), including two heat-labile serotype reference strains, were nonreactive with the commercial antisera, and three strains (4.5%) were typed as serotypes different from those obtained with CDC antisera. Five of the 22 commercial antisera correctly serotyped all homologous strains. Our study indicated that two polyvalent antiserum pools, 7 unabsorbed antisera, and 16 absorbed monovalent antisera are weak and need modification to enhance their antibody titers. Further studies are necessary to explain the antigenic change to a different serotype in three strains.

Animals↗

Detection of rotavirus serotypes G1, G2, G3, and G11 in feces of diarrheic calves by using polymerase chain reaction-derived cDNA probes.

On the basis of antigenic variability in the VP7 outer capsid glycoprotein, at least 14 G serotypes exist for group A rotaviruses. Serotypic diversity exists among bovine rotaviruses (BRV), with serotypes G1, G6, G8, and G10 reported for cattle. Although G1 and G8 rotaviruses were originally described for humans, the recent isolation of G6 and G10 rotaviruses from humans further emphasizes the serotypic similarity between human and bovine rotaviruses and the possible zoonotic potential of rotaviruses. Results of our previous studies have indicated that more than 24% of BRV-positive field samples from diarrheic calves were nonreactive with cDNA probes or monoclonal antibodies to serotypes G6, G8, and G10. In this study, cDNA probes were prepared by polymerase chain reaction amplification of the hyperdivergent regions of the VP7 genes (nucleotides 51 to 392) from human (G1, G2, and G3) and porcine (G4, G5, and G11) rotaviruses. These probes were used in a dot blot hybridization assay to further characterize the G types of 59 BRV strains (fecal samples from diarrheic calves in Ohio, Nebraska, Washington, and South Dakota) that were nonreactive with cDNA probes to G6, G8, and G10. Rotaviruses belonging to serotypes G1 (n = 7), G2 (n = 1), G3 (n = 2), and G11 (n = 3) were identified among the BRV field samples. The BRV associated with these G types accounted for 22% of the samples tested; the other 78% of these samples remained untypeable with these probes. To our knowledge, this is the first report in the United States of the identification among BRV isolates of rotavirus serotypes G1, G2, G3, and G11.

Animals↗

Characterization of endemic Shigella flexneri strains in Somalia: antimicrobial resistance, plasmid profiles, and serotype correlation.

One hundred twelve Shigella flexneri strain isolated from children with diarrheal disease in Somalia in 1983, 1984, 1988, and 1989 were analyzed for serotype, plasmid profile, and genetic location of antimicrobial resistance determinants. The prevalent serotypes were 4 (46% of the isolates), 1b (16%), 2a (16%), 3a (12%), and 6 (8%). Each serotype was associated with a characteristic predominant plasmid profile, whereas no specific correlation between antimicrobial resistance patterns and single serotypes was found. All but three of the strains were resistant at least to ampicillin, chloramphenicol, spectinomycin, and tetracycline. Of these resistant strains, 41 were resistant to sulfonamide and streptomycin and 14 were resistant to trimethoprim or trimethoprim and kanamycin. The genes for resistance to ampicillin, chloramphenicol, spectinomycin, and tetracycline formed a linkage group located on the chromosome of the strains of all serotypes. The genes for resistance to sulfonamide and streptomycin were located on a 6.3-kb plasmid in strains of serotypes 1b, 2a, and 4. Conjugative trimethoprim or trimethoprim and kanamycin resistance plasmids with lengths of 80 to 110 kb were present in strains of serotypes 1b, 2a, 3a, and 4. The systematic presence of a chromosomal component in this uncommon genetic plasmid-chromosome configuration may play a role in the emergence of increased genetic stability of resistance patterns in S. flexneri.

Child↗

Rotavirus serotypes causing severe acute diarrhea in young children in six Australian cities, 1989 to 1992.

Rotavirus VP7 serotypes were identified in stools from 72.9% (1,302/1,784) of hospitalized Australian children in six cities (1989 to 1992) and comprised 1,088 (83.6%) serotype G1 isolates, 84 (6.4%) serotype G2 isolates, 64 (4.9%) serotype G3 isolates, 49 (3.8%) serotype G4 isolates, and 17 (1.3%) isolates of mixed serotypes. The most densely populated cities yielded the greatest diversity of serotypes.

Acute Disease↗

Evaluation of a novel serotyping system for hepatitis C virus: strong correlation with standard genotyping methodologies.

Direct sequencing and analysis of viral genomes are definitive methods for identifying various hepatitis C virus (HCV) genotypes. However, HCV genome sequencing methods are cumbersome and unsuitable for analyzing large numbers of clinical samples. We have developed a convenient, reliable, and reproducible RIBA strip immunoblot assay system for determining HCV serotype. Briefly, the assay consists of an immunoblot strip on which there are five lanes of immobilized serotype-specific HCV peptides from the nonstructural (NS-4) and core regions of the genomes of HCV types 1,2, and 3. HCV serotype is deduced by determining the greatest intensity of reactivity to the NS-4 serotype-specific HCV peptide band in relation to the intensity of the human immunoglobulin G internal control bands on each strip. HCV core peptide reactivity is used only in the absence of NS-4 reactivity. We used this assay to successfully serotype a high percentage of sera from well-documented HCV-infected patients. Our serotyping results correlated 99% with the findings from the standard restriction fragment length polymorphism genotyping methods. Less than 5% of the serum samples were untypeable. For a selected group of alpha interferon-treated patients we observed that the nonresponders (76.2%) and a majority of the responders who relapsed (72.2%) had type 2 HCV infection. A small population (n= 8) of complete responders was split 3:4:1 as type 1, type 2, and type 3, respectively. Our data indicate that this new serotyping assay has the potential to be a highly specific and reliable method for typing of HCV infection in patients.

Genotype↗

Analysis of HL and O serotypes of Campylobacter strains by the flagellin gene typing system.

We recently developed a molecular typing system for Campylobacter jejuni and Campylobacter coli based on restriction fragment length polymorphism analysis of the flagellin gene,flaA (I.Nachamkin, K. Bohachick, and C.M. Patton, J. Clin. Microbiol. 31:1531-1536, 1993). We extended the typing system to 83 flagellin types (designated flaA-1,flaA-2, etc.) on the basis of analysis of 404 isolates of C. jejuni and C. coli including common serotypes isolated in the United States, a selection of less common serotypes, and serotype reference strains. Of the 295 strains previously shown to belong to common HL and O serotypes (C. M. Patton, M.A. Nicholson, S.M. Ostroff, A.A. Ries, I.K. Wachsmuth, and R.V. Tauxe, J. Clin. Microbiol. 31:1525-1530, 1993), six flaA types accounted for 53.6% of strains as follows: flaA-1, 21.7%; flaA-7, 14.9%; flaA-27, 5.1%; flaA-49, 4.4%; flaA-13, 3.7%; and flaA-21, 3.7%. Seventy-five percent of the strains were within 15 flaA types, 90% were within 30 flaA types, and all 295 strains were contained within 52 flaA types. Within each HL or O serotype, there usually were multiple flaA types. For 12 common HL serotypes and 7 common O serotypes, more than 50% of these isolates were a single flaA type. A database was developed by using commercially available restriction fragment length polymorphism analysis software (ProRFLP; DNA ProScan, Inc., Nashville, Tenn.) that should allow other investigators to perform typing with this system.

Animals↗

DNA fingerprinting and serotyping of Campylobacter jejuni isolates from epidemic outbreaks.

The aim of the present investigation was to compare DNA fingerprinting and serotyping (heat-stabile and heat-labile antigens) of isolates from epidemic outbreaks as well as of solitary isolates. Campylobacter jejuni isolates from two epidemic outbreaks in Sweden, one milkborne (35 isolates) and one waterborne (17 isolates), and one waterborne outbreak in Norway (11 isolates), as well as 30 solitary isolates from Swedish patients with gastroenteritis, were analyzed. A total of 93 isolates were analyzed. In the waterborne outbreak in Norway, only one serotype with one DNA pattern was found. In the milkborne outbreak in Sweden, two serotypes (HS2:HL4 and HSNT:HL4) with two different DNA patterns were found. The isolates from the waterborne outbreak in Sweden were different serotypes. For two isolates of the same serotype, different DNA patterns were seen. This was also recorded for isolates from solitary cases. It was concluded that serotyping is a useful tool in most epidemiological situations but sometimes lacks sufficient discriminatory power. DNA fingerprinting can add valuable epidemiological information to that supplied by serotyping and can in some situations provide sufficient epidemiological information when used alone.

Animals↗