RESUMEN

Typhoid fever is endemic in many parts of the world and remains a major public health concern in tropical and sub-tropical developing nations, including Fiji. To address high rates of typhoid fever, the Northern Division of Fiji implemented a mass vaccination with typhoid conjugate vaccine (Vi-polysaccharide conjugated to tetanus toxoid) as a public health control measure in 2023. In this study we define the genomic epidemiology of Salmonella Typhi in the Northern Division prior to island-wide vaccination, sequencing 85% (n=419) of the total cases from the Northern and Central Divisions of Fiji that occurred in the period 2017-2019. We found elevated rates of nucleotide polymorphisms in the tviD and tviE genes (responsible for Vi-polysaccharide synthesis) relative to core genome levels within the Fiji endemic S. Typhi genotype 4.2. Expansion of these findings within a globally representative database of 12 382 S. Typhi (86 genotyphi clusters) showed evidence of convergent evolution of the same tviE mutations across the S. Typhi population, indicating that tvi selection has occurred both independently and globally. The functional impact of tvi mutations on the Vi-capsular structure and other phenotypic characteristics are not fully elucidated, yet commonly occurring tviE polymorphisms localize adjacent to predicted active site residues when overlayed against the predicted TviE protein structure. Given the central role of the Vi-polysaccharide in S. Typhi biology and vaccination, further integrated epidemiological, genomic and phenotypic surveillance is required to determine the spread and functional implications of these mutations.

Asunto(s)

Polisacáridos Bacterianos , Salmonella typhi , Fiebre Tifoidea , Salmonella typhi/genética , Fiji/epidemiología , Fiebre Tifoidea/microbiología , Fiebre Tifoidea/epidemiología , Humanos , Polisacáridos Bacterianos/genética , Heterogeneidad Genética , Vacunas Tifoides-Paratifoides/genética , Genotipo , Mutación , Polimorfismo de Nucleótido Simple , Cápsulas Bacterianas/genética

RESUMEN

The development of strategies for targeting the asymptomatic carriage of Salmonella Typhi in chronic typhoid patients has suffered owing to our basic lack of understanding of the molecular mechanisms that enable the formation of S. Typhi biofilms. Traditionally, studies have relied on cholesterol-attached biofilms formed by a closely related serovar, Typhimurium, to mimic multicellular Typhi communities formed on human gallstones. In long-term infections, S. Typhi adopts the biofilm lifestyle to persist in vivo and survive in the carrier state, ultimately leading to the spread of infections via the fecal-oral route of transmission. In the present work, we studied S. Typhi biofilms directly, applied targeted as well as genome-wide genetic approaches to uncover unique biofilm components that do not conform to the CsgD-dependent pathway as established in S. Typhimurium. We adopted a genome-wide Tn5 mutation screen in S. Typhi in gallstone-mimicking conditions and generated New Generation Sequencing libraries based on the ClickSeq technology to identify the key regulators, IraP and RpoS, and the matrix components as Sth fimbriae, Vi capsule and lipopolysaccharide. We discovered that the starvation sigma factor, RpoS, was required for the transcriptional activation of matrix-encoding genes in vitro, and for S. Typhi colonization in persistent infections in vivo, using a heterologous fish larval model. Overall, our work established a novel RpoS-driven paradigm for the formation of cholesterol-attached Typhi biofilms and emphasized the role(s) of stress signaling pathways for adaptation in chronic infections.

RESUMEN

Introduction: Vaccination with Vi capsular polysaccharide (Vi-PS) or protein-Vi typhoid conjugate vaccine (TCV) can protect adults against Salmonella Typhi infections. TCVs offer better protection than Vi-PS in infants and may offer better protection in adults. Potential reasons for why TCV may be superior in adults are not fully understood. Methods and results: Here, we immunized wild-type (WT) mice and mice deficient in IgG or IgM with Vi-PS or TCVs (Vi conjugated to tetanus toxoid or CRM197) for up to seven months, with and without subsequent challenge with Vi-expressing Salmonella Typhimurium. Unexpectedly, IgM or IgG alone were similarly able to reduce bacterial burdens in tissues, and this was observed in response to conjugated or unconjugated Vi vaccines and was independent of antibody being of high affinity. Only in the longer-term after immunization (>5 months) were differences observed in tissue bacterial burdens of mice immunized with Vi-PS or TCV. These differences related to the maintenance of antibody responses at higher levels in mice boosted with TCV, with the rate of fall in IgG titres induced to Vi-PS being greater than for TCV. Discussion: Therefore, Vi-specific IgM or IgG are independently capable of protecting from infection and any superior protection from vaccination with TCV in adults may relate to responses being able to persist better rather than from differences in the antibody isotypes induced. These findings suggest that enhancing our understanding of how responses to vaccines are maintained may inform on how to maximize protection afforded by conjugate vaccines against encapsulated pathogens such as S. Typhi.

Asunto(s)

Fiebre Tifoidea , Vacunas Tifoides-Paratifoides , Animales , Ratones , Salmonella typhi , Vacunas Conjugadas , Fiebre Tifoidea/prevención & control , Polisacáridos Bacterianos , Inmunoglobulina G , Formación de Anticuerpos , Inmunoglobulina M

RESUMEN

BACKGROUND: We aimed to assess the prevalence of Salmonella Typhi through DNA and IgM-antibody detection methods as a prelude to extended surveillance activities at sites in Ghana, Madagascar, and Ethiopia. METHODS: We performed species-specific real-time polymerase reaction (RT-PCR) to identify bacterial nucleic acid, and enzyme-linked immunosorbent assay (ELISA) for detecting HlyE/STY1498-, CdtB/STY1886-, pilL/STY4539- and Vi-antigens in blood and biopsy specimens of febrile and non-febrile subjects. We generated antigen-specific ELISA proxy cut-offs by change-point analyses, and utilized cumulative sum as detection method coupled with 1000 repetitive bootstrap analyses. We computed prevalence rates in addition to odds ratios to assess correlations between ELISA outcomes and participant characteristics. RESULTS: Definitive positive RT-PCR results were obtained from samples of febrile subjects originating from Adama Zuria/Ethiopia (1.9%, 2/104), Wolayita Sodo/Ethiopia (1.0%, 1/100), Diego/Madagascar (1.0%, 1/100), and Kintampo/Ghana (1.0%, 1/100), and from samples of non-febrile subjects from Wolayita Sodo/Ethiopia (1%, 2/201). While IgM antibodies against all antigens were identified across all sites, prevalence rates were highest at all Ethiopian sites, albeit in non-febrile populations. Significant correlations in febrile subjects aged < 15 years versus ≥ 15 years were detected for Vi (Odds Ratio (OR): 8.00, p = 0.034) in Adama Zuria/Ethiopia, STY1498 (OR: 3.21, p = 0.008), STY1886 (OR: 2.31, p = 0.054) and STY4539 (OR: 2.82, p = 0.022) in Diego/Madagascar, and STY1498 (OR: 2.45, p = 0.034) in Kintampo/Ghana. We found statistical significance in non-febrile male versus female subjects for STY1498 (OR: 1.96, p = 0.020) in Adama Zuria/Ethiopia, Vi (OR: 2.84, p = 0.048) in Diego/Madagascar, and STY4539 (OR: 0.46, p = 0.009) in Kintampo/Ghana. CONCLUSIONS: Findings indicate non-discriminatory stages of acute infections, though with site-specific differences. Immune responses among non-febrile, presumably healthy participants may mask recall and/or reporting bias leading to misclassification, or asymptomatic, subclinical infection signs induced by suppression of inflammatory responses. As most Ethiopian participants were ≥ 15 years of age and not at high-risk, the true S. Typhi burden was likely missed. Change-point analyses for generating ELISA proxy cut-offs appeared robust, though misclassification is possible. Our findings provided important information that may be useful to assess sites prior to implementing surveillance for febrile illness including Salmonella disease.

Asunto(s)

Ácidos Nucleicos , Fiebre Tifoidea , Adolescente , Distrofias Hereditarias de la Córnea , Ensayo de Inmunoadsorción Enzimática , Etiopía/epidemiología , Femenino , Fiebre/microbiología , Ghana/epidemiología , Humanos , Inmunoglobulina M , Madagascar , Masculino , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Salmonella , Salmonella typhi/genética , Fiebre Tifoidea/diagnóstico , Fiebre Tifoidea/epidemiología , Fiebre Tifoidea/microbiología

RESUMEN

Salmonella enterica serovar Typhi causes typhoid fever. It possesses a Vi antigen capsular polysaccharide coat that is important for virulence and is the basis of a current glycoconjugate vaccine. Vi antigen is also produced by environmental Bordetella isolates, while mammal-adapted Bordetella species (such as Bordetella bronchiseptica) produce a capsule of undetermined structure that cross-reacts with antibodies recognizing Vi antigen. The Vi antigen backbone is composed of poly-α-(1→4)-linked N-acetylgalactosaminuronic acid, modified with O-acetyl residues that are necessary for vaccine efficacy. Despite its biological and biotechnological importance, some central aspects of Vi antigen production are poorly understood. Here we demonstrate that TviE and TviD, two proteins encoded in the viaB (Vi antigen production) locus, interact and are the Vi antigen polymerase and O-acetyltransferase, respectively. Structural modeling and site-directed mutagenesis reveal that TviE is a GT4-family glycosyltransferase. While TviD has no identifiable homologs beyond Vi antigen systems in other bacteria, structural modeling suggests that it belongs to the large SGNH hydrolase family, which contains other O-acetyltransferases. Although TviD possesses an atypical catalytic triad, its O-acetyltransferase function was verified by antibody reactivity and 13C NMR data for tviD-mutant polysaccharide. The B. bronchiseptica genetic locus predicts a mode of synthesis distinct from classical S. enterica Vi antigen production, but which still involves TviD and TviE homologs that are both active in a reconstituted S. Typhi system. These findings provide new insight into Vi antigen production and foundational information for the glycoengineering of Vi antigen production in heterologous bacteria.

Asunto(s)

Polisacáridos Bacterianos , Salmonella typhi , Fiebre Tifoidea , Acetiltransferasas/metabolismo , Animales , Polisacáridos Bacterianos/biosíntesis , Polisacáridos Bacterianos/metabolismo , Salmonella typhi/metabolismo , Salmonella typhi/patogenicidad , Fiebre Tifoidea/microbiología , Fiebre Tifoidea/prevención & control , Virulencia

RESUMEN

The extraordinary optical properties of porphyrins have inspired their applications in various fields. Herein, we introduce iron porphyrin bio-mimicked graphene quantum dots (Fe-N-GQDs) as a novel paramagnetic and fluorescent label. The Fe-N-GQD was prepared by the mechanochemical mixing of Fe, N, and C sources followed by pyrolysis at high-temperature and next, the solvothermal treatment was performed. The Fe-N sites in graphene matrix, the structural alterations during the solvothermal treatment, the optical properties, and paramagnetic behaviour were studied using FTIR, Raman and X-ray spectroscopies, and Vibrating sample magnetometer. The structural studies revealed that under solvothermal condition, Fe-N doped graphene sheets cut into ultra-small Fe-N-GQDs containing well-dispersed particles with an average diameter of about 2.5 nm. As a result of Fe-N doping, the photoluminescence quantum yield was enhanced to 86% and strong paramagnetic behaviour was observed. Due to the rich oxygen-containing groups at Fe-N-GQDs surface, it has proper sites for bio-conjugation. The bioconjugated Fe-N-GQDs serve as donors in a prominent fluorescence resonance energy transfer system, while graphene oxide acts as an acceptor. The proposed immunosensor was successfully applied for the detection of Salmonella Typhi Vi antigen in real human serum in the concentration range from 1 pg/mL to 1 µg/mL with the detection limit of 1 pg/mL.

Asunto(s)

Técnicas Biosensibles , Grafito , Porfirinas , Puntos Cuánticos , Humanos , Inmunoensayo , Salmonella typhi

RESUMEN

The development of a nanoparticulate system for the carrier antigen is now an important tool in the vaccination process, since a smaller number of doses is necessary for effective immunization. Thus, in this work a nanoparticulate system using polymers of chitosan and poly (methacrylic acid) (CS-PMAA) to adsorb the Vi antigen of Salmonella Typhi was developed. CS-PMAA nanoparticles with different proportions of chitosan and poly (methacrylic acid) were obtained and reached sizes from 123.9 ± 2.48 to 234.9 ± 2.66 nm, and spherical shapes were seen in transmission microscopy. At pH 7.2, the nanoparticles had a cationic surface charge that contributed to the adsorption of the Vi antigen. Qualitative analyses of the isolated Vi antigen were performed using Fourier-transform infrared spectroscopy, which indicated the presence of all the characteristic bands of the capsular polysaccharide, and nuclear magnetic resonance, which showed signals for the five hydrogens and the N-acetyl and O-acetyl groups which are characteristic of the Vi antigen structure. In the adsorption kinetics study, the Vi capsular antigen, contained in a phosphate buffer solution of pH 7.2, experienced 55% adsorption on the 1-1% (CS-PMAA) nanoparticles. The adsorption kinetics results showed the ability of the nanoparticulate system to adsorb the Vi antigen.

RESUMEN

How Salmonella enterica serovar Typhi (S. Typhi), an important human pathogen, survives the stressful microenvironments inside the gastrointestinal tract and within macrophages remains poorly understood. We report here that S. Typhi has a bonafide stringent response (SR) system, which is mediated by (p)ppGpp and regulates multiple virulence-associated traits and the pathogenicity of the S. Typhi Ty2 strain. In an iron overload mouse model of S. Typhi infection, the (p)ppGpp0 (Ty2ΔRelAΔSpoT) strain showed minimal systemic spread and no mortality, as opposed to 100% death of the mice challenged with the isogenic wild-type strain. Ty2ΔRelAΔSpoT had markedly elongated morphology with incomplete septa formation and demonstrated severely attenuated motility and chemotaxis due to the loss of flagella. Absence of the Vi-polysaccharide capsule rendered the mutant strain highly susceptible to complement-mediated lysis. The phenotypes of Ty2ΔRelAΔSpoT was contributed by transcriptional repression of several genes, including fliC, tviA, and ftsZ, as found by reverse transcriptase quantitative polymerase chain reaction and gene complementation studies. Finally, Ty2ΔRelAΔSpoT had markedly reduced invasion into intestinal epithelial cells and significantly attenuated survival within macrophages. To the best of our knowledge, this was the first study that addressed SR in S. Typhi and showed that (p)ppGpp was essential for optimal pathogenic fitness of the organism.

Asunto(s)

Proteínas Bacterianas/genética , Guanosina Pentafosfato/metabolismo , Interacciones Huésped-Patógeno/genética , Salmonella typhi/genética , Salmonella typhi/patogenicidad , Fiebre Tifoidea/microbiología , Animales , Proteínas Bacterianas/metabolismo , Células CACO-2 , Modelos Animales de Enfermedad , GTP Pirofosfoquinasa/deficiencia , GTP Pirofosfoquinasa/genética , Regulación Bacteriana de la Expresión Génica , Células HT29 , Humanos , Sobrecarga de Hierro/metabolismo , Sobrecarga de Hierro/microbiología , Sobrecarga de Hierro/mortalidad , Sobrecarga de Hierro/patología , Hígado/metabolismo , Hígado/microbiología , Hígado/patología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Polisacáridos Bacterianos/deficiencia , Pirofosfatasas/deficiencia , Pirofosfatasas/genética , Células RAW 264.7 , Salmonella typhi/crecimiento & desarrollo , Salmonella typhi/metabolismo , Transducción de Señal , Bazo/metabolismo , Bazo/microbiología , Bazo/patología , Análisis de Supervivencia , Células THP-1 , Fiebre Tifoidea/metabolismo , Fiebre Tifoidea/mortalidad , Fiebre Tifoidea/patología , Virulencia

RESUMEN

BACKGROUND: Salmonella enterica serovar Dublin is a zoonotic infection that can be transmitted from cattle to humans through consumption of contaminated milk and milk products. Outbreaks of human infections by S. Dublin have been reported in several countries including high-income countries. A high proportion of S. Dublin cases in humans are associated with invasive disease and systemic illness. The genetic basis of virulence in S. Dublin is not well characterized. METHODS: Whole genome sequencing was applied to a set of clinical invasive and non-invasive S. Dublin isolates from different countries in order to characterize the putative genetic determinants involved in the virulence and invasiveness of S. Dublin in humans. RESULTS: We identified several virulence factors that form the bacterial invasome and may contribute to increasing bacterial virulence and pathogenicity including mainly Gifsy-2 prophage, two different type 6 secretion systems (T6SSs) harbored by Salmonella pathogenicity islands; SPI-6 and SPI-19 respectively and virulence genes; ggt and PagN. Although Vi antigen and the virulence plasmid have been reported previously to contribute to the virulence of S. Dublin we did not detect them in all invasive isolates indicating that they are not the main virulence determinants in S. Dublin. CONCLUSION: Several virulence factors within the genome of S. Dublin might contribute to the ability of S. Dublin to invade humans' blood but there were no genomic markers that differentiate invasive from non-invasive isolates suggesting that host immune response play a crucial role in the clinical outcome of S. Dublin infection.

Asunto(s)

Genoma Bacteriano , Salmonella enterica/genética , Salmonella enterica/patogenicidad , Animales , Bovinos , Islas Genómicas/genética , Humanos , Plásmidos , Salmonella enterica/aislamiento & purificación , Virulencia/genética , Factores de Virulencia/genética

RESUMEN

Chitosan has attracted much interest due to its special physical and chemical properties related to drug administration. Nanoparticles delivery systems from Vi Antigen are a promising approach in the struggle against typhoid fever. In this paper, we reported the obtainment and the characterization of Vi Antigen by Infrared spectroscopy as well as Molecular Modeling and Computational Chemistry studies of the Chitosan-Vi Antigen interaction through theoretical models. The results of the theoretical and experimental Infrared spectroscopy showed important bands related to N-Acetyl and O-Acetyl groups present in Vi Antigen. Important interactions related to its adsorption were observed through three-dimensional optimized structures. Two models were proposed for the Chitosan-Vi Antigen in adsorption system, one as a monomer and another as an optimized tetrasaccharide antigen. The Molecular Modeling studies presented the best conformation and binding site on the nanoparticle Chitosan-Vi Antigen in models proposed. Interactions were observed between O-Acetyl and N-Acetyl groups the Vi Antigen and hydroxy, amino and methyl groups the Chitosan.

Asunto(s)

Quitosano/química , Modelos Moleculares , Polisacáridos Bacterianos/química , Simulación del Acoplamiento Molecular , Nanopartículas , Espectroscopía Infrarroja por Transformada de Fourier , Electricidad Estática

RESUMEN

Polysaccharide capsules are surface structures that are critical for the virulence of many Gram-negative pathogenic bacteria. Salmonella enterica serovar Typhi is the etiological agent of typhoid fever. It produces a capsular polysaccharide known as "Vi antigen," which is composed of nonstoichiometrically O-acetylated α-1,4-linked N-acetylgalactosaminuronic acid residues. This glycan is a component of currently available vaccines. The genetic locus for Vi antigen production is also present in soil bacteria belonging to the genus Achromobacter Vi antigen assembly follows a widespread general strategy with a characteristic glycan export step involving an ATP-binding cassette transporter. However, Vi antigen producers lack the enzymes that build the conserved terminal glycolipid characterizing other capsules using this method. Achromobacter species possess a Vi antigen-specific depolymerase enzyme missing in S enterica Typhi, and we exploited this enzyme to isolate acylated Vi antigen termini. Mass spectrometry analysis revealed a reducing terminal N-acetylhexosamine residue modified with two ß-hydroxyl acyl chains. This terminal structure resembles one half of lipid A, the hydrophobic portion of bacterial lipopolysaccharides. The VexE protein encoded in the Vi antigen biosynthesis locus shares similarity with LpxL, an acyltransferase from lipid A biosynthesis. In the absence of VexE, Vi antigen is produced, but its physical properties are altered, its export is impaired, and a Vi capsule structure is not assembled on the cell surface. The structure of the lipidated terminus dictates a unique assembly mechanism and has potential implications in pathogenesis and vaccine production.

Asunto(s)

Aciltransferasas/metabolismo , Proteínas Bacterianas/metabolismo , Lípido A/biosíntesis , Polisacáridos Bacterianos/biosíntesis , Salmonella typhi/metabolismo , Achromobacter/genética , Achromobacter/metabolismo , Aciltransferasas/genética , Proteínas Bacterianas/genética , Ácidos Hexurónicos/metabolismo , Lípido A/genética , Polisacáridos Bacterianos/genética , Salmonella typhi/genética

RESUMEN

Zwitterionic polysaccharides (ZPS) behave like traditional T cell-dependent antigens, suggesting the design of new classes of vaccines alternative to currently used glycoconjugates and based on the artificial introduction of a zwitterionic charge motif onto the carbohydrate structure of pathogen antigens. Here we report the new synthesis and antigenic evaluation of di-/tri-saccharide fragments of Salmonella typhi Vi polysaccharide, as well as of their corresponding zwitterionic analogues. Our strategy is based on versatile intermediates enabling chain elongation either by iterative single monomer attachment or by faster and more flexible approach using disaccharide donors. The effect of structural modifications of the synthetic compounds on antigenic properties was evaluated by competitive ELISA. All the oligosaccharides were recognized by specific anti-Vi polyclonal antibodies in a concentration-dependent manner, and the introduction of a zwitterionic motif into the synthetic molecules did not prevent the binding.

Asunto(s)

Amino Azúcares/síntesis química , Disacáridos/síntesis química , Polisacáridos Bacterianos/química , Trisacáridos/síntesis química , Amino Azúcares/química , Amino Azúcares/inmunología , Animales , Anticuerpos/inmunología , Disacáridos/química , Disacáridos/inmunología , Ensayo de Inmunoadsorción Enzimática , Caballos , Polisacáridos Bacterianos/inmunología , Salmonella typhi , Relación Estructura-Actividad , Trisacáridos/química , Trisacáridos/inmunología

RESUMEN

An injectable Vi-capsular polysaccharide vaccine against typhoid fever is available but vaccine-induced immunity tends to wane over time. The phenomenon of immunotolerance or hyporesponsiveness has earlier been described for polysaccharide vaccines such as pneumococcal capsular polysaccharide vaccine and some publications also suggest a possible immunotolerance after revaccination with Vi-capsular polysaccharide vaccines. In this study, post-immunisation antibody concentrations in adult travellers first vaccinated with a Salmonella typhi Vi-capsular polysaccharide vaccine (primary vaccination group) were compared with those having received one or more vaccinations previously (multiple vaccinations group). Vaccines administered were Typherix(®) (GlaxoSmithKline), Typhim Vi(®) (Sanofi Pasteur MSD) or Hepatyrix(®) (GlaxoSmithKline). Blood samples were obtained prior to vaccination (day 0) and on day 28 (-1/+14) after vaccination. Serum Vi-Antigen IgG concentrations were measured by ELISA. Of the 85 subjects included in the per protocol data set, 45 (53%) belonged to the multiple vaccinations group. In both groups, geometric mean antibody concentrations (GMCs) were significantly higher after vaccination than before vaccination. Pre-vaccination GMCs were lower in the primary vaccination group than in the multiple vaccinations group (3.40 µg/ml versus 6.13 µg/ml, P=0.005), while there was no significant difference in the post vaccination GMCs between groups (11.34 µg/ml versus 14.58 µg/ml, P=0.4). In the multiple vaccinations group, vaccination was performed 18 to 57 months after the last vaccination (median 38 months) and there was a negative correlation between time since last vaccination and antibody concentration on day 0. In conclusion, we were not able to demonstrate a relevant immunotolerance after multiple versus primary vaccination with S. typhi Vi-capsular polysaccharide vaccines.

Asunto(s)

Anticuerpos Antibacterianos/sangre , Inmunización Secundaria , Polisacáridos Bacterianos/inmunología , Vacunas contra la Salmonella/administración & dosificación , Vacunas contra la Salmonella/inmunología , Adolescente , Adulto , Anciano , Ensayo de Inmunoadsorción Enzimática , Femenino , Voluntarios Sanos , Humanos , Tolerancia Inmunológica , Inmunoglobulina G/sangre , Memoria Inmunológica , Masculino , Persona de Mediana Edad , Viaje , Adulto Joven

RESUMEN

This experiment was conducted to assess the efficacy of typhoid vaccine newly produced by purifying Vi antigen of Salmonella typhi. With Karber method, LD50 of challenging organism (S. typhi ty2) was determined as 6.31 CFU/mouse, and then the organism was used for the study. With Probits method, ED50 of the vaccine was determined as 0.016 microgram / 0.5 ml / mouse. The ELISA titer (0.5097+/-0.0606) was 4 times in the group treated with high dose (0.25 microgram/0.5ml) as in control (0.1113+/-0.0110). Six major protein bands of 66, 55, 35, 33, 18, and 9 kd were detected in Western blot analysis with serum of a vaccine treated mouse, whereas only one weak band of about 35 kd was detected with serum of a control mouse. We concluded that typhoid vaccine produced by purifying Vi antigen of S. typhi very effectively prevent S. typhi infection in mice.

Asunto(s)

Animales , Masculino , Ratones , Anticuerpos Antibacterianos/inmunología , Antígenos Bacterianos/inmunología , Western Blotting , Ensayo de Inmunoadsorción Enzimática , Dosificación Letal Mediana , Modelos Logísticos , Ratones Endogámicos BALB C , Polisacáridos Bacterianos/inmunología , Salmonella typhi/química , Fiebre Tifoidea/inmunología , Vacunas Tifoides-Paratifoides/administración & dosificación

RESUMEN

BACKGROUND: Typhoid fever is diagnosed by culture or serological study. The confirmative diagnosis of typhoid fever is made by culture of the causative orga-nism usually from body fluids. Serological test is a supportive diagnostic tool, which is useful for early dia-gnosis. In Severance Hospital, Vi-indirect fluorescent antibody test(Vi-IFAT) using the Vi-antigen of Salmo-nella typhi has been used in the diagnosis of typhoid fever since 1989. We investigated the test results from the past 7 years, in order to clarify the sensitivity and specificity of Vi-IFAT. METHODS: A retrospective study was done on pa-tients whose chief complaint was fever and who were tested using Vi-IFAT in the Severance Hospital from 1989 to 1996. The positive value for Vi-IFAT was de- fined as 1:64 or higher. RESULTS: The sensitivity and specificity of Vi-IFAT for typhoid fever was 94.4% and 95.1%, respectively. The positive and negative predictive values were 85.7% and 98.2% respectively. Positive rates of Vi-IFAT after fever onset increased with time and 68% were positive before the first week. From the first to the second week, 89.5% were positive and after the second week, 100% were positive. CONCLUSION: Vi-IFAT is not only a valuable sero-logic test for the diagnosis of typhoid fever, but also useful in the early diagnosis of the disease.

Asunto(s)

Líquidos Corporales , Diagnóstico , Diagnóstico Precoz , Fiebre , Estudios Retrospectivos , Sensibilidad y Especificidad , Pruebas Serológicas , Fiebre Tifoidea