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Detection of IgM antibody against region IV flagellin of Salmonella paratyphi A.

Salmonella paratyphi A is a pathogenic bacterium that causes paratyphoid fever. The current laboratory diagnostic techniques are unsatisfactory. To improve diagnosis, a plasmid (pSK-8E) encoding phase 1 flagellin gene nucleotide position 452-890 from S. paratyphi A has been constructed. The recombinant protein expressed from the plasmid has been used to develop an indirect ELISA for IgM antibody detection. Sera from patients with hemoculture positive for S. paratyphi A, S. typhi, other gram-positive and gram-negative bacteria, and dengue hemorrhagic fever as well as from healthy control subjects were tested. Sensitivity, specificity, positive and negative predictive values of the test were 56.9%, 98.8%, 90.6% and 92.1%, respectively. Since the sensitivity was low, the explanation for this result was investigated. It was found that the sensitivity of the test could be increased to 83.3% if the sera were obtained 9-12 days after onset of fever. The sera obtained earlier or later gave only 33.3% and 66.6% sensitivity, respectively. This result suggests that the IgM antibody detection assay which we have developed is a valuable tool for diagnosis of S. paratyphi A infection when the serum samples are taken at the appropriate time.

Antibodies, Monoclonal↗

Identification of a new site for ferrichrome transport by comparison of the FhuA proteins of Escherichia coli, Salmonella paratyphi B, Salmonella typhimurium, and Pantoea agglomerans.

The fhuA genes of Salmonella paratyphi B, Salmonella typhimurium, and Pantoea agglomerans were sequenced and compared with the known fhuA sequence of Escherichia coli. The highly similar FhuA proteins displayed the largest difference in the predicted gating loop, which in E. coli controls the permeability of the FhuA channel and serves as the principal binding site for the phages T1, T5, and phi80. All the FhuA proteins contained the region in the gating loops required in E. coli for ferrichrome and albomycin transport. The three subdomains required for phage binding were contained in the gating loop of S. paratyphi B which is infected by the E. coli phages, whereas two of the subdomains were deleted in S. typhimurium and P. agglomerans which are resistant to the E. coli phages. Small deletions in a surface loop adjacent to the gating loop, residues 236 to 243 and 236 to 248, inactivated E. coli FhuA with regard to transport of ferrichrome and albomycin, but sensitivity to T1 and T5 was fully retained and sensitivity to phi80 and colicin M was reduced 10-fold. Full-size FhuA hybrid proteins of S. paratyphi B and S. typhimurium displayed S. paratyphi B FhuA activity when the hybrids contained two-thirds of either the N- or the C-terminal portions of S. paratyphi B and displayed S. typhimurium FhuA activity to phage ES18 when the hybrid contained two-thirds of the N-terminal region of the S. typhimurium FhuA. The central segment of the S. paratyphi B FhuA flanked on both sides by S. typhimurium FhuA regions conferred full sensitivity only to phage T5. The data support the essential role of the gating loop for the transport of ferrichrome and albomycin, identified an additional loop for ferrichrome and albomycin uptake, and suggest that several segments and their proper conformation, determined by the entire FhuA protein, contribute to the multiple FhuA activities.

Amino Acid Sequence↗

Proposal of Salmonella paratyphi sp. nov., nom. rev. and request for an opinion to conserve the specific epithet paratyphi in the binary combination Salmonella paratyphi as nomen epitheton conservandum.

We propose Salmonella paratyphi sp. nov., nom. rev., by raising Salmonella choleraesuis subsp. choleraesuis serovar Paratyphi A to species status and request an Opinion to include the specific epithet paratyphi in the binary combination of Salmonella paratyphi in the list of epitheta specifica conservanda.

Salmonella↗

The pathogenicity of strains of Salmonella paratyphi B and Salmonella java.

AIMS: To relate the diseases caused by strains of Salmonella paratyphi B and S. java to pathogenic mechanisms expressed by these bacteria for the purpose of organism discrimination. METHODS AND RESULTS: Epidemiological data relating to cases of disease caused by strains of S. paratyphi B and S. java, isolated over a 10-year period, were analysed with respect to patients' symptoms, particularly those involving enteric fever. Strains of S. paratyphi B and S. java were also examined for a range of known pathogenic mechanisms. Infection with S. paratyphi B involved pyrexia in 12.5% of patients compared with 2.2% of patients infected with S. java. These organisms could not be differentiated based on the pathogenic properties examined. CONCLUSIONS: Strains of S. paratyphi B appear not to be a major cause of enteric fever but primarily a cause of gastroenteritis, in common with S. java. Both organisms express similar pathogenic mechanisms, and strains of S. java are probably d-tartrate utilizing variants of S. paratyphi B. SIGNIFICANCE AND IMPACT OF THE STUDY: Strains of S. paratyphi B are very closely related organisms, primarily causing gastroenteritis. From this study it would appear that strains of S. paratyphi B are not a major cause of enteric fever.

Adolescent↗

Utilization of d-tartaric acid by Salmonella paratyphi B and Salmonella java: comparison of anaerobic plate test, lead acetate test and turbidity test.

d-Tartrate dehydrase of Salmonella java is an oxygen-sensitive enzyme active in cultures incubated under the poorly aerated conditions of static culture but not in fully aerated shaken cultures nor on plates incubated aerobically. On plates of d-tartrate minimal agar incubated anaerobically the enzyme or the degradation products of d-tartrate are exported from d-tartrate-positive cells and are available to d-tartrate-negative bacteria. This may give misleading growth results when d-tartrate-positive and d-tartrate-negative strains are tested for growth on the same plate of d-tartrate minimal agar. The lead-acetate test terminated at 24 h, the 24 h turbidity test and the ability to grow on d-tartrate minimal agar within 48 h differentiated 53 S. paratyphi B strains that were negative in each of the three tests from 76 S. java that were positive in each of the tests. An intermediate group of eight strains utilized d-tartrate in Difco bacto-peptone water to give a positive lead acetate reaction at 2 days, were stimulated to a varying degree by d-tartrate in Oxoid peptone water within the same period of incubation and grew poorly on d-tartrate minimal agar. These latter strains may be deficient in a permease controlling uptake of d-tartrate or export of d-tartrate dehydrase. Inability to utilize d-tartrate is unlikely to be the single character accountable for the reputed enhanced pathogenicity of S. paratyphi B when compared with S. java. Indications for the existence of an enzyme, complementary to and mutually exclusive with d-tartrate dehydrase, that has a positive correlation with pathogenicity are discussed.

Anaerobiosis↗

Salmonella paratyphi A is more genetically homogeneous than Salmonella typhi, as indicated by pulsed-field gel electrophoresis.

We analyzed 18 Salmonella Paratyphi A and 12 Salmonella Typhi isolates from domestic and imported cases in Aichi, Japan, using pulsed-field gel electrophoresis. Paratyphoid fever cases have increased and outbreaks of Salmonella Paratyphi A occasionally occur in Japan, but S. Paratyphi A has not been extensively analyzed. Our study suggests significant genetic homogeneity among Salmonella Paratyphi A belonging to different phage types, which is in contrast to the genetic heterogeneity of Salmonella Typhi. These results suggest that a limited number of clones are responsible for paratyphoid fever.

Bacteriophage Typing↗

Immunochemical relations of salmonella paratyphi C with the salmonellae of group B.

Agar-gel precipitations of proteins from S. paratyphi C, S. paratyphi B and S. typhimurium against homologous and heterologous antibacterial sera, prepared in rabbits, demonstrated a strong relatedness between these species belonging to different serogroups. The findings explain and substantiate previous experiments in which high cross-protections were obtained in groups of mice immunized with proteins from these species and subsequently infected with S. typhimurium or its "in vivo" related S. paratyphi B and S. paratyphi C.

Agglutination↗

Genotypic typing and phylogenetic analysis of Salmonella paratyphi B and S. java with IS200.

Salmonella paratyphi B and Salmonella java are biovars of common serotype 1,4,[5],12:b:1,2 which respectively cause human paratyphoid fever and gastroenteritis. In order to define genotypes and phylogenetic relationships in this group, we examined representative strains for restriction fragment length polymorphisms (RFLPs) in and around the 16S ribosomal RNA (rrn) genes, and the five to eleven insertion sites of the Salmonella-specific DNA insertion sequence IS200. One of four 16S rrn profiles was predominant, and was shared by the majority of strains, irrespective of their designation as S. paratyphi B or S. java. On the other hand, thirteen unique IS200 profiles were found and this technique was able to distinguish, for the first time, distinct genotypes for S. paratyphi B and S. java. One of the S. paratyphi B profiles, Spj-IP1.0, represented a globally-distributed clone. Greater diversity was detected within IS200 profiles of S. java than within those of S. paratyphi B. IS200 profiles described a phylogenetic complex in which strains of both biovars could be placed. They constituted reproducible molecular fingerprints, which could be compared in a band-matching database suitable for molecular epidemiological typing.

Bacterial Typing Techniques↗