Capsular Polysaccharide Antigen Research Articles

Typing of human isolates of Streptococcus agalactiae (group B streptococcus, GBS) strains from Zimbabwe.

Serotyping and genotyping are important tools in epidemiological studies of group B streptococcal (GBS) infections, which are important diseases in man, particularly in newborns. In the present study, 241 GBS isolates from Zimbabwe, comprising 124 carrier isolates from pregnant women and 117 isolates from patients hospitalised for various diseases, were serotyped. Antibodies specific for the capsular polysaccharide antigens (CPAs) Ia, Ib and II-V and antibodies specific for the surface-localised proteins, c(alpha), c(beta), R1, R3 and R4 were used for serotyping. Strains of the CPA types Ia (17%), III (47.7%) and V (23.2%) predominated. Of the various protein antigens, c(alpha) and R4 were expressed with highest frequency, c(alpha) by 100% of the CPA type Ia strains and R4 by 92% of the CPA type III strains. The R3 protein occurred frequently (24%), especially in type V strains (84%). A total of 25 serovariants was detected in the strain collection with the variants Ia/c(alpha) (16%), III/R4 (43.5%) and V/c(alpha), R3 (14.1%) occurring with the highest frequency. Serotype and subtype distribution of the carrier isolates were essentially similar to those of the disease-associated isolates. Genomic heterogeneity was demonstrated by pulsed-field gel electrophoresis of type III/R4 and type V/c(alpha), R3 isolates, but to a much lesser extent than recorded with Norwegian strains. These results demonstrate that many variants of GBS occur in the Zimbabwean population. The data obtained may assist in the formulation of a possible future GBS vaccine for Zimbabwe and perhaps for other African countries.

Clinical and immunological evaluation of patients with mild IgG1 deficiency.

Serum IgG subclass concentrations were determined in patients visiting, the pulmonology out-patient clinic with chronic respiratory tract problems. A total of 24 patients with a serum IgG1 concentration < 4.9 g/l (i.e. below the reference range) and normal values for IgG2, IgM and IgA were included. Patients with a selective IgG1 deficiency were vaccinated with a 23-valent pneumococcal polysaccharide vaccine. There were nine patients with a poor antibody response to pneumococcal capsular polysaccharide antigens. Responsiveness to protein antigens was intact in all patients. Patients with pneumonia showed a significantly lower anti-polysaccharide response in the IgG2 subclass than patients without pneumonia. Patients with recurrent sinusitis showed a significantly lower response in the IgA isotype after vaccination with pneumococcal polysaccharide vaccine compared with non-sinusitis patients. It can be concluded that patients with recurrent sinopulmonary infections and a mild IgG1 subclass deficiency have an impaired IgG1 anti-polysaccharide response, which can extend to decreased IgG2 and IgA anti-polysaccharide responses.

Humoral immune responses following experimental infection of goats with Mycoplasma capricolum subsp. capripneumoniae

Goats housed in microbiologically secure facilities were experimentally endobronchially infected with Mycoplasma capricolum subsp. capripneumoniae (Mccp), causal agent of contagious caprine pleuropneumonia (CCPP). The animals were monitored over an 8-week period post-infection (p.i.). Elevated temperatures were observed 2–7 days p.i., reaching a maximum of 41.5°C in one animal (1884). By 8 weeks p.i. the infection was successfully cleared, with no Mccp being recovered from the lungs, serum or nasal passages. Mccp was not isolated from serum throughout the experiment, either directly by culture or indirectly via polymerase chain reaction (PCR). Humoral immune responses against Mccp capsular polysaccharide (CPS) were generally poor when measured by ELISA. CPS antigen was present in the serum of all infected animals early in the infection (day 14 p.i.), although in one animal (1855) CPS antigen persisted throughout. This was the only animal to exhibit a serious cough (day 5–19 p.i.). Successful diagnosis of CCPP was achieved using two different types of latex agglutination test (CPS antibody and CPS antigen detection test), immunoblotting and a blocking ELISA, although the latter lacked sensitivity until later in the infection (35–40 days p.i.). Only a single animal (1855) was detected positive using the current complement fixation test (CFT). Strong immune responses to protein antigens were detected by IgG and IgM immunoblotting from the first time point at day 14 p.i. IgM immunodominant bands of 220, 85, 62 and 40 kDa were observed in the 3 infected animals and from CFT-positive CCPP field sera. Band intensity gradually diminished throughout the experiment. IgG immunodominant bands of 108, 70, 62, 44, 40 and 23 kDa were shared between experimentally-infected and field sera, with band intensity either remaining unchanged or increasing from day 14 p.i. These bands were not present using pre-infection sera. Of the diagnostic tests used, only the CPS antibody detection latex agglutination test and IgG immunoblotting gave positive diagnoses throughout the entire period post-infection (days 14–53 p.i.).

Detection of Streptococcus pneumoniae Antigen by a Rapid Immunochromatographic Assay in Urine Samples

Evaluation of a newly available rapid (15 min) immunochromatographic membrane test (ICT) to detect Streptococcus pneumoniae in urine samples, in order to assess its utility in the diagnosis of bacteremic and nonbacteremic pneumococcal pneumonia. Retrospective study. We studied urine samples from 51 patients with bacteremic and nonbacteremic pneumonia due to S pneumoniae diagnosed by blood culture and pneumococcal polysaccharide capsular antigen detection by counterimmunoelectrophoresis in urine samples, 16 patients with probable pneumococcal pneumonia, 71 patients with nonpneumococcal pneumonia, and 16 patients with pneumonia but no pathogen identified. Urine samples were collected and frozen at - 20 degrees C until used. The ICT test was performed following the instructions of the manufacturer. S. pneumoniae antigen was detected in 41 of 51 patients with pneumococcal pneumonia (80.4%); results were positive in 23 of 28 bacteremic cases (82.1%) and in 18 of 23 nonbacteremic cases (78.3%). From patients with a diagnosis of presumptive pneumococcal pneumonia, antigen was detected in seven urine samples (43.7%) and also in one case of the 16 patients with pneumonia but no pathogen identified. The specificity of the ICT test was 97.2%. The ICT assay is a valuable tool for the diagnosis of pneumococcal pneumonia, especially for the nonbacteremic cases.

Influence of Bispecific Antibodies on the In Vitro Bactericidal Activity of Bovine Neutrophils Against Staphylococcus aureus

We conducted the following study to determine if bispecific antibodies enhance the bactericidal activity of bovine neutrophils. Bispecific antibodies were synthesized by chemically crosslinking bovine neutrophil monoclonal antibodies to Staphylococcus aureus 305 capsule polysaccharide monoclonal antibodies. The efficiency of chemically coupling monoclonal antibody monomers was approximately 50% for each bispecific antibody produced. Monoclonal antibodies against neutrophils enhanced the respiratory burst activity of neutrophils by 2.3- to 2.5-fold. To determine the influence of bispecific antibodies on neutrophil function, S. aureus 305 was preincubated with various concentrations of bispecific antibodies and neutrophils were then added to the opsonized bacteria at different bacteria to neutrophil ratios. The bactericidal activity of neutrophils was expressed as a percentage reduction in colony-forming units in test cultures compared with the number of colony-forming units in control test cultures that did not contain bispecific antibodies or neutrophils. The addition of bispecific antibodies to test cultures increased the bactericidal activity of neutrophils. A reduction in colony-forming units as a function of increasing the S. aureus 305 to neutrophils ratio was observed in both the absence and presence of bispecific antibodies. However, a greater reduction was observed in the presence of bispecific antibodies. Increasing concentrations of bispecific antibodies enhanced the bactericidal activity of neutrophils at a constant S. aureus 305 to neutrophil ratio of 1:500. The results indicate that bispecific antibodies that recognize both S. aureus 305 capsular polysaccharide and neutrophil antigens potentiate the bactericidal activity of neutrophils.

Prophylactic and therapeutic efficacy of antibodies to a capsular polysaccharide shared among vancomycin-sensitive and -resistant enterococci.

Enterococci are important nosocomial pathogens that are increasingly difficult to treat due to intrinsic and acquired resistance to antibiotics, including vancomycin. A recently described capsular polysaccharide (CP) isolated from Enterococcus faecalis 12030 was used to evaluate the potential efficacy of active or passive immunotherapy regimens as adjunctive treatments. Evaluation of protective efficacy was carried out in immunocompetent mice challenged intravenously (i.v.) with live enterococci. In nonimmune mice, i.v. inoculations resulted in high levels of bacteria in kidneys, spleens, and livers 5 days after challenge. Mice immunized with four 10-microg doses of CP antigen/mouse were protected against challenge with the homologous E. faecalis strain. High-titer opsonic immunoglobulin G was also induced by immunizing rabbits with the purified CP, and passive transfer of this antiserum to mice produced significantly lower bacterial counts in organs than did normal rabbit serum or sterile saline. Antibodies to the polysaccharide isolated from E. faecalis 12030 were protective against Enterococcus faecalis OG1RF and against two serologically related, vancomycin-resistant Enterococcus faecium clinical isolates. Antibodies to this CP antigen were also effective as a therapeutic reagent in mice when passive therapy was initiated 48 h after live bacterial challenge. These data indicate that CP antigens from enterococci are potential targets of protective antibodies and that these antibodies may be useful for prophylaxis and treatment of enterococcal infections.

Prevalence, capsular type distribution, anthropometric and obstetric factors of group B Streptococcus (Streptococcus agalactiae) colonization in pregnancy.

To establish the prevalence, serotype distribution, anthropometry and obstetric factors of Group B Streptococcus (GBS) colonization in pregnant women. Cross sectional survey. Chinhoyi General Hospital. 206 pregnant women attending the antenatal clinic at Chinhoyi General Hospital were systematically randomly sampled. All the isolates were serotyped on the basis of capsular polysaccharide (CHO) antigen designated, Ia, Ib, II, III, IV and V. 65 (31.6%) were carriers of GBS. The serotypes found were, type III (41.8%), type V (37.4%), type Ia (11.0%), type IV (3.3%), type Ib (3.3%) type II (1.0%) and 2.0% of the isolates were non-typable. All isolates were sensitive to penicillin and resistant to gentamycin. Colonization was more common in women with parity 0 to 2 (4.6%) and age group 20 to 24 years (43.1%). There was some evidence (p = 0.063) to suggest that GBS was more often isolated from the vagina (12.6%) than from the rectum (6.3%). There was a high prevalence of GBS colonization among pregnant women in Chinhoyi. Types III and V were the most common serotypes found.

Capsular polysaccharide and O-specific antigen divergently modulate pulmonary neutrophil influx in an Escherichia coli model of gram-negative pneumonitis in rats.

Enteric gram-negative bacilli cause a severe, often life-threatening pneumonia. An improved understanding of the pathogenesis of this infection may lead to improved treatment. Nearly all of the responsible gram-negative bacilli possess capsular polysaccharides and/or an O-specific antigen as part of their lipopolysaccharide (LPS). We hypothesized that these surface polysaccharides may modulate the pulmonary host response. To investigate this, a rat pneumonitis model was used, and pulmonary neutrophil influx, a critical aspect of host defense, was measured. To assess for the effect of the capsule and O-specific antigen on this host response, three proven, isogenic derivatives that are deficient in capsular polysaccharide alone (CP9.137), the O-specific antigen moiety of the LPS alone (CP921), and both the capsular polysaccharide and O-specific antigen (CP923), as well as their wild-type parent (CP9), were used as challenge strains at various intratracheal challenge inocula (CI). Total lung myeloperoxidase (MPO), a surrogate marker for neutrophils, was measured for 15 h post-bacterial challenge. To determine the effect of capsule and the O-specific antigen on the measured MPO levels, a mathematical model was developed and used to describe the MPO levels as a function of time for each CI of each of the four strains. The results from this analysis demonstrated that in the absence of the K54 capsule, 80.7 times the CI is necessary to achieve the same maximum MPO level relative to K54 positive strains (P < 0.0001). In contrast, a diametric effect was observed in the absence of the O-specific antigen, where 0.13 times the CI was necessary to achieve the same maximum MPO level relative to O4-positive strains (P = 0.0032). No interactive effect was observed between the capsule and the O-specific antigen. These findings demonstrate that these surface polysaccharides modulate pulmonary neutrophil influx and suggest that the K54 capsular polysaccharide is a proinflammatory mediator and that the O4-specific antigen attenuates the proinflammatory response. If these speculations are substantiated, an understanding of how the capsule and the O-specific antigen modulate host response could have significant therapeutic implications. The potential use of biologic modulators directed against the host response, as well as approaches based on inactivating bacterial components (e.g., surface polysaccharides) in attempts to modify sepsis syndromes, could be developed.

The immunostimulating activities of anti-tumor polysaccharide from K1 capsular (polysaccharide) antigen isolated from Klebsiella pneumoniae

We have previously shown that K1 capsular polysaccharide antigen (K1CPS) of Klebsiella exhibits anti-tumor activities. In the present study, we examined the effect of K1CPS on cytotoxic effector cells. We found that K1CPS could activate many cytotoxic effector cells including alloreactive cytotoxic T cells and tumor-infiltrating lymphocytes (TILs). Moreover, K1CPS could increase the anti-tumor activity of lymphokine-activated killer (LAK) cells, both in vitro and in vivo. The i.p. injection of K1CPS in low dose could enhance the LAK cytotoxicity and the effect was further potentiated by coculture of LAK cells with K1CPS and low concentration of murine rIL-2 in vitro. The phenotypic characterization revealed that K1CPS might contribute to the increase in CD3+ LAK cell subpopulation by its in vivo priming effect. In addition, the K1CPS-treated LAK cells were able to inhibit the growth of WEHI-164 tumor cells in vivo in Winn-type inhibition assay. Subcutaneous (s.c.) and intraperitoneal (i.p.) adoptive infusion of LAK cells (splenocytes from K1CPS-treated WEHI-164-bearing mice cultured with K1CPS-plus-rIL-2) into WEHI-164 sarcoma-bearing mice could slightly cause regression in terms of tumor diameter, and more significantly in sarcoma weight.

Effects of adjuvants on the immune response of staphylococcal alpha toxin and capsular polysaccharide (CPS) in rabbit.

This study was performed to isolate a vaccine strain of S. aureus from clinical or subclinical mastitis and to choose the most optimal adjuvant for immune response of alpha toxin and capsular polysaccharide (CPS) of field strain. Of thirty strains of S. aureus isolated from milk of clinical or subclinical mastitis, V112 strain isolated from milk of gangrenous mastitis was used in this vaccine. Twenty one of rabbits were allocated into 5 groups based on adjuvants and immunized twice every 2 weeks for 8 weeks. This vaccine was composed of alpha toxin (10 hemolytic units) and formalinized whole cells (1 x 10(11) cells/ml. Five rabbits received PBS solution as a control group. The highest antibody titers against alpha toxin and CPS were observed in dextran sulfate- and aluminium hydroxide-adjuvant group at 8 weeks after immunization, respectively. These results of the study showed that one adjuvant could not induce strong and long-term immune response of alpha toxin and CPS antigens. Therefore, the use of combined adjuvants in subunit vaccine may be useful and feasible.

Influence of the culture conditions on extracellular lyase activity related to K5 polysaccharide

The K5 capsular polysaccharide antigen of some Escherichia coli strains is the non-sulphated precursor in heparin biosynthesis. It is composed by two components, 16 000 and 1500 Da, whose ratio depends on the activity of the extracellular form of a lyase synthesized by the same K5 producer strain. The lyase activity on the K5 chain size was greatly influenced by the medium composition employed for the lyase production. The control of lyase activity results in defined ratios of the two components in the K5 polysaccharide that is suitable for the semisynthetic production of heparin-like molecules.

Detection of pneumolysin in sputum.

Western blot detection of the species-specific pneumococcal product, pneumolysin (SPN), was shown to be almost as sensitive as PCR for the non-cultural detection of pneumococci in 27 Streptococcus pneumoniae culture-positive sputa from patients stated to have chest infections. Both techniques were considerably more sensitive than counter-current immuno-electrophoresis for pneumococcal capsular polysaccharide antigens (CPS-CIE) on the same specimens. Sensitivities for PCR, SPN-immunoblotting and CPS-CIE were 100%, 85% and 67%, respectively. In 11 S. pneumoniae culture-negative sputa taken from patients receiving antibiotics, but with proven recent pneumococcal infection, PCR and SPN-blot were positive in six (in two of which CPS-CIE was also positive), PCR alone was positive in one and SPN-blot alone was positive in one. In 11 S. pneumoniae culture-negative samples from patients not receiving antibiotics, all three tests were negative in eight, PCR was positive in three (in one of which CPS-CIE was also positive), but SPN-blot was negative in all 11. In 16 S. pneumoniae culture-negative samples from patients receiving antibiotics and with no known recent pneumococcal infections, one or more non-cultural test was positive in 11. Although further evaluation is required to assess the significance of pneumolysin detection in relation to carriage and infection and to devise a more suitable test format, these preliminary studies suggest that pneumolysin detection is a promising new approach to the non-cultural diagnosis of pneumococcal chest infection.

Synthesis and preclinical evaluation of glycoconjugate vaccines against group B Streptococcus types VI and VIII.

Group B Streptococcus (GBS) types VI and VIII are prevalent among serotypes isolated from pregnant women in Japan. Maternal vaccination with a safe and effective GBS vaccine has been proposed as a rational approach to prevent neonatal GBS disease. Because antibody specific for the capsular polysaccharide (CPS) antigens of GBS is protective, vaccines were developed with purified type VI and VIII CPS coupled to tetanus toxoid. In rabbits the newly synthesized conjugate vaccines elicited high-titered, type-specific antibody that was opsonically active in vitro. Moreover, litters born to mice actively vaccinated with the conjugate vaccines, in contrast to uncoupled CPS or saline, were protected against an ordinarily lethal challenge of GBS of homologous serotype. GBS types VI and VIII conjugate vaccines of the design presented may be important components of a multivalent GBS vaccine for use in regions where these serotypes predominate.

Crystallization and preliminary X-ray diffraction analysis of antigen-binding fragments which are specific for antigenic conformations of sialic acid homopolymers.

Meningococcal meningitis is a severe childhood disease which often results in significant disability or death. Two major etiological agents of meningitis are the group B meningococci and capsular type K1 E. coli. The virulence of these organisms is attributable to structural mimicry between their common alpha(2-8)-polysialic acid capsular polysaccharide and human tissue antigens, which allows the bacteria to evade immune surveillance. There is currently no effective vaccine to protect against this infection. It has been demonstrated that the capsular polysaccharide of the bacteria can adopt a unique 'antigenic conformation'. This antigenic conformation has formed the basis for the development of an N-propionylated polysialic acid vaccine. Immunization trials in mice with this vaccine show the production of two groups of antibodies, of which only N-propionylated polysialic acid-specific were protective. Knowledge of the structure of the antigen-binding site which recognizes the protective epitope is essential to determining the antigenic conformation of the polysaccharides, and is a critical aspect in understanding and improving the action of potential vaccines. The antigen-binding fragments (Fab) of one protective (13D9) and one non-protective (6B9) monoclonal antibody specific for the capsular polysaccharides of group B meningococci have been crystallized and have undergone preliminary X-ray diffraction analysis. Both crystals are observed to scatter X-rays to approximately 1.7 A resolution at the A1 station at the Cornell High-Energy Synchrotron Source. 13D9 has an orthorhombic unit cell with a = 41.8, b = 102.3, c = 134.7 A, with space group P212121. Fab 6B9 has an orthorhombic unit cell with a = 89.6, b = 132.0 and c = 36.9 A, with space group P21212.

Re-evaluation of the serotypes of Serratia marcescens and separation into two schemes based on lipopolysaccharide (O) and capsular polysaccharide (K) antigens.

Chemical and serological analysis has revealed that many of the 29 O serotype reference strains of Serratia marcescens contain both neutral and acidic polysaccharides which correspond to LPS O antigens and capsular K antigens, respectively. New O and K antigen typing schemes have therefore been devised, based on the known chemical structures of the surface polysaccharides of the organism. These schemes were designed to allow the specific detection of these antigens on unknown strains using ELISAs. O antigens were detected using whole cells cultured in broth then autoclaved to remove capsular material, while K antigens were detected using formolized whole cells which had been cultured on glycerol agar to enhance capsule production. After testing with the 29 reference strains as well as 423 distinct clinical strains, it was apparent that different aspects of chemical structure were associated with different degrees of serological reactivity and the typing schemes were modified further to accommodate this. In general, the O antigen repeating unit structures were chemically simple with di- or trisaccharide backbones. Serological specificity was often provided solely by the presence or absence of an O-acetyl substituent, or a change in the linkage between two sugar residues. Five of the O serotypes in the new scheme were represented by 12 of the 29 reference strains, while three reference strains lacked O antigens altogether, resulting in the elimination of 10 of the original O types. In contrast, the K antigen repeating unit structures were more complex and chemically diverse, having at least four sugar residues. Three K types were each seen in two reference strains while 12 of the 29 reference strains were acapsular. Thus, the resulting schemes contain 19 O types and 14 K types and allow the definitive serotype identification of S. marcescens.

Detection of human serum antibody to encapsulated strains of Staphylococcus aureus by enzyme-linked immunosorbent assay inhibition test

A specific and rapid enzyme-linked immunosorbent assay (ELISA) inhibition test was employed for detection of immunoglobulins to Staphylococcus aureus (S. aureus) capsular polysaccharide in human serum. Capsular polysaccharide antigens obtained from Smith diffuse (capsular type 2), Reynolds (capsular type 5), or Becker (capsular type 8) strains of S. aureus were added to microplates coated with these strains. Seventy-four patients with open fractures (31 serum samples from those with staphylococcal infections, 10 serum samples from those with non-staphylococcal infections, and 33 serum samples from the non-infected group) and 28 serum samples from healthy controls were then added. The plates were incubated at 37°C for 2 h and the ELISA was performed. The ELISA inhibition assay showed remarkable inhibition with the capsular type 2, 5, and 8 polysaccharides in the 33 serum samples from the non-infected group and in the 28 serum samples from the healthy controls, but low inhibition was observed with the 31 sera with staphylococcal infections. Positive immunoglobulin (Ig)G and IgM titers showed marked inhibition with this assay, but IgA titer were not seen in any samples. These results indicate that the quantitation of human serum antibody against S. aureus capsular polysaccharide by the ELISA inhibition assay is useful for the demonstration of protective activities against S. aureus.

Cervical Secretions in Pregnant Women Colonized Rectally with Group B Streptococci have High Levels of Antibodies to Serotype III Polysaccharide Capsular Antigen and Protein R

Group B streptococci (GBS) colonizing the female genital tract will often infect newborn infants during delivery. In 200 pregnant women studied, 14% were colonized with GBS in the cervix, 12% in the rectum, and 9% in both cervix and rectum. We have previously reported that antibody levels to GBS serotypes Ia, II, and III in sera and cervical secretions were increased in women colonized in the rectum and/or cervix, when analyzed by a whole-cell ELISA. Here, we report the levels of antibodies to GBS serotype III capsular polysaccharide antigen (CPS III) and to protein antigen R4, which are present in most GBS III strains. Compared to culture-negative women, the group of women colonized rectally had markedly elevated levels of immunoglobulin (Ig)A and IgG antibodies in cervical secretions to both CPS III and protein R4 (P < 0.01 and P < 0.001, respectively). In sera, the corresponding differences between culture-negative and culture-positive women were less pronounced, or not present. In contrast to antibody levels to whole-cell GBS, antibody levels to CPS III and protein R4 in cervical secretions were not significantly increased in women colonized only in the cervix, except that IgA antibodies to protein R4 were slightly elevated (P < 0.05). These findings suggest that capsular type-specific polysaccharides and protein R4 in a mucosal vaccine might induce protective antibodies against GBS colonization of the uterine cervix.

Comparison of IgG IgGl lgG2 immune responses to pneumococcal polysaccharide in atopic nonatopic children

The levels of naturally occurring IgG, IgGl and lgG2 antibodies against polyvalent pneumococcal capsular polysaccharide antigen (Pneumovax®) were compared between atopic and nonatopic children with different ages, 6–12 months and 1, 2, 3, 4, 5–9 and 10–15 years, by enzyme-linked immunosorbent assay. Children with asthma, atopic dermatitis, food allergy or a combination of these, and those having serum IgE levels exceeding 50 IU/mL at 6–12 months old and 100 IU/mL at more than 1-year-old were included as atopic groups. Asymptomatic children whose serum IgE levels were less than these atopic standards and those not having detectable IgE antibodies to Dermatophagoides farinae comprised the nonatopic groups. Geometric mean levels of IgG and IgG 1 antibodies against pneumococcal antigen increased steadily with age, and that of IgG2 antibodies was low until 3 years of age and then gradually increased age-dependently up to 15 years of age. The levels of IgG antibody as well as IgG 1 and IgG 2 antibodies were not significantly different between atopic and nonatopic children in any age group. This suggests that the immune response to the most common bacterial pathogen in the respiratory tract does not influence atopic status.

Low sensitivity of counter-current immuno-electrophoresis for serodiagnosis of typhoid fever.

Counter-current immuno-electrophoresis was evaluated as a diagnostic test for the serodiagnosis of typhoid fever with somatic (O), flagellar (H) and capsular polysaccharide (Vi) antigens of Salmonella typhi on the sera of patients who were blood culture positive (confirmed typhoid cases) or had high Widal agglutination titres, > or = 320, (presumptive typhoid cases). Of the 37 sera from confirmed cases, 30% showed positivity with O antigen, 24% with H antigens and 51% with Vi antigen. In patients with a presumptive diagnosis, 45% were positive for O antibody, 27% for flagellar antibody and 52% for Vi antibody. When all three antigens were combined the reactivity to any of the antigens was found to be 59% in confirmed typhoid cases, 79% in presumptive typhoid cases and 93% in patients who were simultaneously positive by blood culture and Widal agglutination. However, none of the sera from 45 controls gave a positive precipitation reaction with any of the antigens. It is concluded that counter-current immuno-electrophoresis is a rapid test with low sensitivity and high specificity with Vi antigen, a panel of antigens being most effective, and is, therefore, recommended for rapid diagnosis of typhoid fever.

Streptococcus agalactiae beta gene and gene product variations.

Streptococcus agalactiae (group B streptococci; GBS) are serotyped on the basis of the capsular polysaccharide antigens and subtyped on the basis of the strain-variable and surface-localised c proteins c alpha, c beta, and R proteins. This study compared c beta protein detection and the polymerase chain reaction (PCR) for beta gene detection, by examining 50 clinical GBS strains. The c beta protein was detected by antibody-based immunofluorescence in a GBS whole-cell assay and Western blotting by probing with the anti-c beta antibody or human IgA. Absorption experiments were performed to test for surface-anchoring of c beta; and bacterial supernates were examined to test for c beta production. Primers for the PCR target regions resulted in a 620-bp product that included beta gene-encoding IgA-binding domains. The results demonstrated four categories of GBS with respect to the beta gene and the c beta protein: (1) strains (16 of 50) that harboured the beta gene and regularly expressed normal surface-localised c beta with a M(r) of 120 kDa; (2) strains (5 of 50) that harboured the gene but did not express the protein; (3) strains (2 of 50) that harboured the gene but expressed a c beta that was not surface-localised and had reduced M(r); (4) strains (27 of 50) without beta gene and c beta expression. One strain amongst the third group generated a PCR product of 1330 bp. These results demonstrate considerable strain variability of the beta gene of GBS and of its product the c beta protein.