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Serotype-specific immunoglobulin G antibody responses to pneumococcal polysaccharide vaccine in children with sickle cell anemia: effects of continued penicillin prophylaxis.

OBJECTIVES: (1) To determine serotype-specific IgG antibody responses to reimmunization with pneumococcal polysaccharide vaccine at age 5 years in children with sickle cell anemia and (2) to determine whether continued penicillin prophylaxis had any adverse effects on these responses. STUDY DESIGN: Children with sickle cell anemia, who had been treated with prophylactic penicillin for at least 2 years before their fifth birthday, were randomly selected at age 5 years to continue penicillin prophylaxis or to receive placebo treatment. These children had been immunized once or twice in early childhood with pneumococcal polysaccharide vaccine and were reimmunized at the time of randomization. RESULTS: Serotype-specific IgG antibody responses to reimmunization varied according to pneumococcal serotype but in general were mediocre or poor; the poorest response was to serotype 6B. The antibody responses were similar in subjects with continued penicillin prophylaxis or placebo treatment, and in subjects who received one or two pneumococcal vaccinations before reimmunization. The occurrence of pneumococcal bacteremia was associated with low IgG antibody concentrations to the infecting serotype. CONCLUSIONS: Reimmunization of children with sickle cell anemia who received pneumococcal polysaccharide vaccine at age 5 years induces limited production of serotype-specific IgG antibodies, regardless of previous pneumococcal vaccine history. Continued penicillin prophylaxis does not interfere with serotype-specific IgG antibody responses to reimmunization.

Adult↗

Serotypes of Escherichia coli isolated from healthy infants in Berlin, Germany and Melbourne, Australia.

The characterization of Escherichia coli strains isolated from healthy infants under one year of age with respect to O:H serotype, K1 and K5 antigens in two disparate parts of the developed world was the purpose of this investigation. A total of 450 strains were examined, 264 from Berlin and 186 from Melbourne. Of all the 220 different O:H serotypes found, 179 were only isolated once, 90 in Berlin and 89 in Melbourne. However, 30 of the 41 O:H serotypes (73.2%) found more than once were isolated in both centers. The most commonly identified serotypes were found in both centers and included O1:H-; O1:H7; O2:H2; O2:H4; O2:H7; O4:H5; O6:H-; O6:H1; O15:H1; O18:H7; O25:H1; and 075:H-. Potentially pathogenic serotypes were found in both cities. Enteropathogenic E. coli (EPEC)-associated serotypes (O18:H7; O26:H-; O44:H34; O86:H-; O128:H2) were present in 11 cases and enterohaemorrhagic E. coli (EHEC)-associated types including O26:H11; O128:H2) were present in four cases. The distributions of serotypes found were similar in the two cities, strongly suggesting the wider applicability of these results.

Berlin↗

Characterization and evaluation of some virulence markers of Listeria monocytogenes strains isolated from Brazilian cheeses using molecular, biochemical and serotyping techniques.

A total of 207 L. monocytogenes strains isolated from different types of cheeses commercialized in the city of Rio de Janeiro, Brazil, were serotyped and evaluated for their ability to produce beta-haemolysin and lecithinase and to adsorb Congo red dye. Of the 207 strains, 59.9, 27.5 and 12.6% belonged to serotypes 1/2a, 1/2b and 4b, respectively. In addition, 175 strains of L. monocytogenes produced lecithinase while strains of the other species did not. Some of the non-L. monocytogenes strains adsorbed the dye Congo red, while some L. monocytogenes did not. Statistical analysis of the results showed significant differences (P < 0.05) amongst the virulence tests and the three serotypes found. In the present study, 32 L. monocytogenes strains were also analyzed by RAPD (randomly amplified polymorphic DNA). RAPD analysis allowed the discrimination among strains of different serotypes, as well as among strains of the same serotype. It is important to emphasize that the use of more than one primer is needed for characterization of L. monocytogenes strains. With RAPD the strains were grouped into six different profiles, some of them common for strains belonging to different serotypes. The results also indicated a close genetic relationship among strains of different serotypes.

Bacteriological Techniques↗

A comparison between methods of identification and serotyping of encapsulated strains of Haemophilus influenzae.

Seven methods of serotyping of Haemophilus influenzae were evaluated. Comparing slide agglutination, staphylococcal coagglutination, latex agglutination, counterimmunoelectrophoresis, immunofluorescence, capsular swelling, and cultivation on antiserum agar the commercial coagglutination test was most reliable, most rapid, and easiest to perform. To identify all six serotypes this coagglutination test had to be combined with slide agglutination. With most methods best results were achieved by using cultures incubated at 37 degrees C for 6 h. As nonencapsulated strains often agglutinated unspecifically, selection of probably typeable strains was useful. Differentiation with help of colonial morphology and opalescent growth was facilitated by cultivation on Brain Heart Infusion (BHI) Chocolate Agar and testing of growth factor requirements on translucent BHI Agar with strips containing the growth factors V, X, and VX, respectively. In broth turbid growth was a hint for encapsulation. Nigrosin staining, a negative capsule staining, proved to be useful if specific antisera are not available. From 252 clinical isolates of H. influenzae 216 were not typeable. 36 strains could be serotyped. 27 (75%) belonged to serotype b, 6 (16.6%) were serotype e, 3 (8.3%) were serotype f. Serotype e and f were most difficult to identify. Spectrum of patients and diseases were corresponding to the findings of other authors. Less well-known infections like cellulitis (erysipelas of the cheeks) and arthritis were observed, too. Rapid identification of at least H. influenzae type b could render treatment in some cases more effective by early application of a suitable antibiotic.

Agglutination Tests↗

Frequency of Pseudomonas aeruginosa serotypes in burn wound infections and their resistance to antibiotics.

Pseudomonas aeruginosa plays a prominent role as an etiological agent involved in serious infections in burned patients. In this study P. aeruginosa infections were analyzed at the Motahari Burn Center in Tehran (from 22 December 1998 to April 1999) to estimate their frequency, antibiotic susceptibility and serotypes. One hundred and eighty-four positive cultures and 205 bacterial strains were isolated among swabs or biopsy specimens during the study period. Pseudomonas was found to be the most common (57%) followed by Acinetobacter (17%), Escherichia coli (12%), Staphylococcus aureus (8%) and other organisms (6%). The frequency of P. aeruginosa resistance to gentamicin, ceftizoxime, carbenicillin, cephalothin and ceftazidime was over 90%. The antibiotics to which P. aeruginosa was most sensitive were amikacin and tetracyclin. The "O" serotypes isolated from the 117 Pseudomona aeroginosa isolates were serotypes O:2, O:5, O:6, O:8, O:11, O:12 and O:16. The most common serotype was O:6 (20/17%) followed by O:11 (18/15%) and O:5 (14/12%). The serotype most resistant was O:16 (8%) and the most sensitive was O:8 (2%). Since treatment of infection with available antibiotics according to the results attained proved to be difficult, prevention of infection in the burned patients is considered as an appropriate means of conquering overcoming infection problems. The sum of frequencies of serotypes O:6, O:11, O:5 and O:16 was more than 60%, therefore vaccination of burn patients with polyvalent antiserum to these serotypes could possibly produce immunity in more than half of the burned patients.

Acinetobacter↗

Investigations on the species specificity of Mannheimia (Pasteurella) haemolytica serotyping.

Eleven serotypes (1, 2, 5-9, 12-14 and 16) have been demonstrated within Mannheimia haemolytica. Subsequent serotyping of 166 Mannheimia haemolytica-like strains, genetically and phenotyphically distinct from Mannheimia haemolytica, and isolated from ruminants, pigs, hares and rabbits showed that 13.2% were typeable, 19 of which were serotype 11 representing strains now being classified as M. glucosida. In addition, three strains belonged to serotypes 6, 9 and 16, respectively. Additionally, the serotyping results of 98 (P.) haemolytica-like isolates from non-ruminant sources collected by the UK Veterinary Investigation Centres during the period 1982-1996 were investigated. None of these isolates have been kept, making further genetic characterization impossible. Among these isolates, 25.5% were typeable representing serotypes 1, 2, 3, 5, 6, 9, 10, 13 and 15. Substantial evidence has been reported indicating that M. haemolytica-like isolates from non-ruminant sources represent species different from M. haemolytica. The present investigation underlines that serotyping does not represent a reliable method for the identification of M. haemolytica or M. glucosida. These observations emphasize that extended phenotypic and genetic characterization is necessary for the proper identification of these organisms.

Animals↗

Identification of Shigella serotypes by restriction of amplified O-antigen gene cluster.

Due to the scarcity of distinctive biochemical reactions for differentiation of Shigella-Escherichia coli, antigenic analysis has long been used for identification and typing of Shigella isolates. Nevertheless, several intra- and interspecific cross-reactions have been reported to disturb serotyping assays. Shigella serotyping is also occasionally affected by the transition from the smooth (S) form to the rough (R) form. Thus, there is a need for the development of novel robust and discriminating methods for Shigella identification and typing. Characteristically, all genes specifically involved in O-antigen synthesis are clustered in E. coli, Shigella, and Salmonella. Published oligonucleotide sequences complementary to JUMPstart and gene gnd, the conserved flanking sequences upstream and downstream of O-antigen gene clusters, were used to amplify the O-antigen gene cluster of representative strains of each Shigella serotype. A unique, amplified fragment was generally observed for each serotype (size ranging from 6 kbp to 17 kbp). Clearly identifiable and reproducible patterns were obtained for each serotype after MboII digestion of the products, except for S. boydii 12 which showed two distinct patterns, and S. flexneri serotypes 1 to 5 and X and Y which showed a single pattern. A database was built with the Taxotron package allowing automated identification of clinical Shigella isolates to all known serotypes.

Bacterial Proteins↗

Comparison of hepatitis C virus serotyping and genotyping in French patients.

BACKGROUND: Hepatitis C virus (HCV) serotyping has been proposed as an alternative assay to determine the respective genotype as it is more rapid, simple and less expensive than polymerase chain reaction (PCR) based typing methods. OBJECTIVES: A serotyping assay was compared with a genotyping assay to determine the infecting hepatitis C virus type in chronically HCV infected patients eligible for interferon therapy. STUDY DESIGN: An enzyme immunoassay (HC01, Murex) was tested to identify HCV types 1, 2 and 3 specific antibodies in 134 PCR-positive sera from chronically infected patients which had been previously genotyped by a reverse hybridization assay (INNO-LiPA HCV I, Innogenetics). Respectively nine and seven sera were from HIV-seropositive and hemodialysis patients. Unreactive sera and those with discrepant results were retested by a new version (HC02) extended to types 4, 5 and 6. RESULTS: The distribution frequency of HCV genotypes was subtype 1a, 16.4%; subtype 1b, 46.3%; subtype 2a, 7.5%; subtype 3a, 20.9%; type 4, 4.5%; type 5, 0.7%; and co-infections, 3.7%. Among all the patients, 95% were of type 1, 2 or 3. The antibody reactivities of hemodialysis (1/7; P < 0.05) and HIV-seropositive patients (4/9; P = 0.06) were lower than for the patients seen at the hepatology unit (87/118). For these latter patients, the serotyping assay was interpretable in 71% and concordant in 64% of the samples with the genotyping assay. Out of the 84 samples with interpretable results, 75 sera were correctly serotyped (89% specificity). The two mixed results obtained by serotyping did not correspond to genotype co-infections (n = 3) and reciprocally. Six discrepancies were ruled out by the new assay, but the 2 untypeable sera remained unsolved, and four out of six sera with genotype 4 were serotyped as type 5. CONCLUSIONS: Serotyping could be an attractive approach if the reactivity was improved and the subtyping possible.

Adult↗

Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers.

The distribution of the Escherichia coli attaching and effacing (eae) gene in strains of verotoxin-producing E. coli (VTEC) isolated from cattle and humans was studied. The majority of strains isolated from humans with bloody diarrhoea or HUS and cattle with severe diarrhoea were eae positive (82 and 83% respectively). In contrast, 59% of VTEC isolated from asymptomatic cattle were eae negative and of the remaining 41% that were eae positive, the majority were serotype O157. H7. The nucleotide sequence of the 3' end of the eae gene of enteropathogenic E. coli (EPEC) of serotype O55. H7 was found to be almost identical to that of serotype O157. H7. Specific primers are described which detect the eae sequences of VTEC serotypes O157. H7, O157. H-, and EPEC serotypes O55. H7 and O55. H-. The nucleotide sequence of the 3' end of the eae gene of serotype O111. H8 differed significantly from that of O157. H7. Primers were developed to specifically identify the eae sequences of VTEC serotypes O111. H- and O111. H8. We conclude that whereas the majority of VTEC associated with disease in cattle and humans possess the eae gene, the gene itself may not be necessary to produce haemorrhagic colitis and HUS. Sequence heterogeneity in the 3' end of eae alleles of VTEC permits specific identification of subsets of these organisms.

Adhesins, Bacterial↗

Acquisition and invasiveness of different serotypes of Streptococcus pneumoniae in young children.

Rates of acquisition and mean duration of nasal carriage of different serotypes of Streptococcus pneumoniae have been estimated by fitting a stochastic model to longitudinal carriage data in children from Papua New Guinea. Immunogenicity and two indices of relative invasiveness were determined for each serotype. Immunogenic serotypes were less frequently acquired and were carried for shorter periods, but no relationship between immunogenicity and invasiveness was apparent using either index of invasiveness. Frequent invasion was associated with a high acquisition rate and high frequency and prolonged duration of carriage. Carriage studies can provide a broad indication of which serotypes cause invasive disease but not the proportion of disease due to individual serotypes; some serotypes which cause invasive disease (e.g. serotype 46) are not found even in extensive carriage studies. The antibiotic resistance of carriage organisms, however, does approximate the resistance patterns of invasive organisms and thus may be used to monitor changing patterns of antimicrobial susceptibility in the community.

Antibodies, Bacterial↗

The influence of competition and vaccination on the coexistence of two pneumococcal serotypes.

Streptococcus pneumoniae (pneumococcus) is one of the most important bacterial pathogens and a leading cause of mucosal infections (e.g. otitis media) and various forms of serious diseases (e.g. pneumonia, meningitis, bacteraemia) in developing and developed countries. Based on the polysaccharide capsule, there are at least 90 different pneumococcal serotypes, which may compete with each other to colonize the nasopharynx. Newly developed protein-polysaccharide conjugated vaccines have been shown to provide protection against disease caused by the serotypes included in the vaccine, and also against colonization (carriage). It is feared that yet uncommon, but nonetheless pathogenic serotypes which have been suppressed by competition, may become more prevalent in carriage and disease after large-scale use of conjugate vaccines. In this paper, we use transmission models of pneumococcal carriage to study how competition and vaccination influence the coexistence of two serotypes. According to our results, direct (physical) competition between two pneumococcal serotypes only influences colonization if the duration of naturally acquired immunity is short. By contrast, indirect (antibody-mediated) competition is of influence only if naturally acquired immunity is long lasting. Vaccination reduces the prevalence of the target serotype--an effect that is enforced by the presence of directly competing bacteria. The emergence of a non-target serotype after vaccination is only observed if bacteria compete directly. These results emphasize the importance of studying whether bacteria compete directly or indirectly and for how long people are protected in order to assess the long-term effects of sero-competition.

Carrier State↗

Prevalence and serotyping of Porphyromonas gingivalis in an Indonesian population.

In this study, the prevalence and serotype distribution of Porphyromonas gingivalis in an Indonesian population (n=158) is described. The relationship between subgingival P. gingivalis and periodontal attachment loss was investigated. The serotype distribution and periodontal parameters were studied. Serotyping was also used to study person-to-person transmission between siblings and between spouses. Approximately 50% of the subjects had periodontal attachment loss > or =3 mm at 1 or more recorded sites. The population was divided into 2 subgroups based on the presence or absence of P. gingivalis. No differences in plaque index, pocket depth, clinical attachment loss, bleeding upon probing, % of sites with > or =3 mm clinical attachment loss and % of sites with probing pocket depth > or =5 mm, were observed between both sub-populations. All known 6 capsular serotypes were found in the study population, with the exception of the K1 type. Detection of 1 of the known P. gingivalis serotypes was not related with the amount of clinical attachment loss. In 3 out of 29 sibships with more than one member positive for P. gingivalis, an identical P. gingivalis serotype was found. None of the 15 couples in the study shared an identical P. gingivalis serotype, indicating that transmission is probably not a common phenomenon in this population.

Adolescent↗

Actinobacillus actinomycetemcomitans in Chinese adults. Serotype distribution and analysis of the leukotoxin gene promoter locus.

The aim of the present study was to examine the Actinobacillus actinomycetemcomitans carrier rate in Chinese subjects, and to determine serotype distribution, presence of the leukotoxin gene lktA and the structure of the lktA-promoter region. Subgingival microbiological samples were obtained from 31 Chinese subjects with moderate to advanced adult periodontitis, 73 young factory workers, and 81 adult residents of a rural area. Bacterial isolates phenotypically identified as A. actinomycetemcomitans were found in 116 of the 185 subjects (detection frequency over-all: 63%). Presence of the leukotoxin gene lktA was demonstrated for all 115 isolates that could be subcultured. The PCR analysis of the lktA-promoter region showed that none of these strains had the deletion in the promoter region known to enhance expression of lktA. No significant difference in the frequency of A. actinomycetemcomitans could be observed between the subjects of the 3 study groups. Analysis by logistic multiple regression indicated a homogeneous distribution of A. actinomycetemcomitans in the 3 cohorts and a lack of significant influence of subject gender or age. Serotype a was found in 21 subjects, serotype b was found in 9, serotype c in 67 and serotype e in 11 individuals. Serotype d was not detected in any subject. Nontypeable isolates, lacking serotype a, b, c, d, or e antigens, were found in 9 individuals. A high prevalence irrespective of gender, age, and cohort suggests that A. actinomycetemcomitans is a common constituent of the normal flora in the Chinese subjects of this study and suggests differences in the microbiological composition of subgingival plaque may exist for this population group as compared to north American and European populations.

Adolescent↗

Serotypes and clinical manifestations of group B streptococcal infections in western Sweden.

OBJECTIVES: To study the serotype distributions of group B streptococci (GBS) isolated from blood and cerebrospinal fluid and from the genital tract of pregnant women and to investigate any possible relation between serotype, age and clinical manifestation. METHODS: Invasive strains were collected from 1988 to 1997 and genital strains from 1995 to 1996. Strains of GBS were serotyped with coagglutination. Clinical data were obtained from hospital notes. RESULTS: A total of 144 invasive strains, 78 from neonates and infants and 66 from adults, were serotyped. The most common isolates from neonates and infants were types III (62%), Ia (18%), and V (9%). The most common isolates from adults were types III (29%), Ib (23%), V (21%) and II (15%). A majority of the adults (94%) had an underlying medical condition. The most common serotypes of the 114 strains isolated from the genital tract of pregnant women were types III (32%), V (22%), Ia (13%), Ib (13%) and II (11%). CONCLUSIONS: Serotype III was the single most frequent GBS isolate from infants and adults. Serotype V, which appeared first in 1992, was the third most frequent isolate. A vaccine containing five GBS capsular polysaccharides appears to be appropriate for the Swedish population.

Adult↗

Use of molecular subtyping in surveillance for Salmonella enterica serotype typhimurium.

BACKGROUND: Because Salmonella enterica serotype typhimurium is the most common serotype isolated from persons with salmonellosis in the United States, it is difficult to detect unusual clusters or outbreaks. To determine whether molecular subtyping could be useful in public health surveillance for S. enterica serotype typhimurium, the Minnesota Department of Health initiated the routine use of pulsed-field gel electrophoresis (PFGE) of isolates. METHODS: Beginning in 1994, all S. enterica serotype typhimurium isolates submitted by clinical laboratories to the Department of Health were subtyped by PFGE. A standard questionnaire was used to interview patients about possible sources of infection. RESULTS: From 1994 through 1998, 998 cases of infection with S. enterica serotype typhimurium were reported to the Minnesota Department of Health (4.4 cases per 100,000 person-years). PFGE was performed on 958 of the isolates (96 percent), and 174 different patterns were identified. Sixteen outbreaks with a common source were identified, accounting for 154 cases. PFGE subtyping made it possible to confirm 10 outbreaks that involved small numbers of cases in institutional settings. Of six larger, community-based outbreaks, four would probably not have been recognized without PFGE subtyping. These four outbreaks accounted for 96 of the 154 culture-confirmed outbreak cases (62 percent). Fifty-six of 209 isolates tested for antimicrobial susceptibility (27 percent) were resistant to at least five antimicrobial agents. The multidrug-resistant isolates identified had unique PFGE patterns. CONCLUSIONS: Routine molecular subtyping of S. enterica serotype typhimurium by PFGE can improve the detection of outbreaks and aid in the identification of multidrug-resistant strains. Combining routine molecular subtyping with a method of rapid communication among public health authorities can improve surveillance for S. enterica serotype typhimurium infections.

DNA, Bacterial↗

Serotypes and antimicrobial susceptibility of group B streptococcus isolated from neonates in Casablanca.

The serotypes and the levels of antibiotic resistance of 59 Streptococcus agalactiae isolates from neonates in Casablanca, from February 1992 to July 1997, were studied. Most of the isolates (86.4%) were recovered from early-onset disease. The serotype distribution was as follows: serotype III 39%; serotype Ia 32.2%; and serotype V 10.2%. All strains were susceptible to penicillin G, cefotaxime and ampicillin, whereas 1 strain was resistant to erythromycin. No high level of resistance to gentamicin was detected. A vaccine should comprise the most prevalent serotypes and also provide protection against serotype V disease. The antibiotic susceptibility patterns reported here support the recommended treatment and prophylaxis of invasive group B Streptococcal disease.

Drug Resistance, Microbial↗

Serotyping and quantitative determination of in vitro antibiotic susceptibility of Actinobacillus pleuropneumoniae strains isolated in Belgium (July 1991-August 1992).

One hundred and ninety nine Actinobacillus pleuropneumoniae isolates obtained from the respiratory tract of pigs in Belgium (1991-1992) were serotyped using slide agglutination and slide precipitation tests. Biotype 1-serotype 2 and 3 strains were isolated most frequently and represented 36% and 21% of the isolates, respectively. Serotype 9 and serotype 5B strains represented 15% and 10% of the total number of isolates, respectively, while serotypes 7, 8, 10 and 11 were isolated only sporadically. Serotypes 5B, 10 and 11 were isolated for the first time in Belgium. Biotype 2 strains represented 12.5% of the total number and all belonged to the serotype 2. The in vitro susceptibility of 138 strains was determined for ten commonly used antibiotics. For three antibiotics, ampicillin, chloramphenicol, and oxytetracycline, a bimodal distribution of strain sensitivities was observed. A total of 17 strains (12%) were considered to have acquired resistance to one or more of these antibiotics.

Actinobacillus Infections↗

Molecular analysis of the genetic variation among penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae serotypes in Canada.

Few studies have examined the genetic relatedness within and between serotypes of Streptococcus pneumoniae. Penicillin-susceptible S. pneumoniae (PSSP) isolates (n=170) and penicillin-resistant S. pneumoniae (PRSP) isolates (n=48) belonging to 13 different serotypes from across Canada were studied to further understand the molecular epidemiology of S. pneumoniae. PRSP of serotypes 6B, 14, and 19F appeared to be novel clones, differentiated by pulsed-field gel electrophoresis from the recognized international penicillin-resistant clones belonging to these serotypes. In addition, not all PRSP strains shared similar genetic backgrounds with PSSP strains of the same serotype, suggesting that in Canada, certain PRSP strains have emerged from new unique lineages of PRSP. Although there was significant heterogeneity among S. pneumoniae of different serotypes, certain serotypes (3, 7F, and 22F) appeared to be more clonally related despite having originated from different geographic regions.

Cluster Analysis↗