RESUMEN
Twenty-eight febrile Sri Lankan patients with undiagnosed fever for 7 days after hospital admission, who responded to empirical treatment with doxycycline, were retrospectively investigated using microimmunofluorescence assay to verify whether they had rickettsial infection. Eleven (39%) patients were confirmed as having spotted fever group rickettsioses and 10 (36%) as having Orientia tsutsugamushi. Seven were negative for all tests. This suggests that greater use of doxycycline appears justified for patients with undiagnosed fever in settings where rickettsial diseases are endemic or re-emerging with inadequate diagnostic facilities.
Asunto(s)
Antibacterianos/uso terapéutico , Doxiciclina/uso terapéutico , Fiebre de Origen Desconocido/microbiología , Infecciones por Rickettsia/complicaciones , Rickettsia/aislamiento & purificación , Adulto , Femenino , Fiebre de Origen Desconocido/tratamiento farmacológico , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Orientia tsutsugamushi/aislamiento & purificación , Infecciones por Rickettsia/diagnóstico , Infecciones por Rickettsia/tratamiento farmacológico , Tifus por Ácaros/diagnóstico , Sri Lanka , Adulto JovenAsunto(s)
Anaplasma/aislamiento & purificación , Ehrlichia/aislamiento & purificación , Garrapatas/microbiología , Animales , Técnicas Bacteriológicas/métodos , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , ADN Ribosómico/química , ADN Ribosómico/genética , Perros/parasitología , Caballos/parasitología , Panamá , Perisodáctilos/parasitología , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Prevalencia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Rickettsia conorii, the etiologic agent of Mediterranean spotted fever is widely distributed in Southern Europe, the Middle East, Africa, India and the Caspian region. In the Mediterranean region, the brown dog tick, Rhipicephalus sanguineus, is the recognized vector of R. conorii. To study tick-pathogen relationships and pathogenesis of infection caused in model animals by the bite of an infected tick, we attempted to establish a laboratory colony of Rh. sanguineus persistently infected with R. conorii. Rhipicephalus sanguineus ticks of North American and Mediterranean origin were exposed to R. conorii isolates of African (R. conorii conorii strain Malish) and Mediterranean (R. conorii israelensis strain ISTT) origin. Feeding of ticks upon infected mice and dogs, intra-hemocoel inoculation, and submersion in suspensions of purified rickettsiae were used to introduce the pathogen into uninfected ticks. Feeding success, molting success and the longevity of molted ticks were measured to assess the effects of R. conorii on the survival of Rh. sanguineus. In concordance with previously published results, Rh. sanguineus larvae and nymphs from both North American and Mediterranean colonies exposed to R. conorii conorii Malish experienced high mortality during feeding and molting or immediately after. The prevalence of infection in surviving ticks did not exceed 5%. On the other hand, exposure to ISTT strain had lesser effect on tick survival and resulted in 35-66% prevalence of infection. Rh. sanguineus of Mediterranean origin were more susceptible to infection with either strain of R. conorii than those from North America. Previous experimental studies had demonstrated transovarial and transstadial transmission of R. conorii in Rh. sanguineus; however, our data suggest that different strains of R. conorii may employ different means of maintenance in nature. The vertebrate host may be a more important reservoir than previously thought, or co-feeding transmission between different generations of ticks may obviate or lessen the requirement for transovarial maintenance of R. conorii.
Asunto(s)
Rhipicephalus sanguineus/microbiología , Rickettsia conorii/fisiología , Animales , Vectores Artrópodos/microbiología , Fiebre Botonosa/microbiología , Fiebre Botonosa/transmisión , Reservorios de Enfermedades/microbiología , Perros , Larva/microbiología , Ratones , Ninfa/microbiología , Conejos , Rickettsia conorii/aislamiento & purificaciónRESUMEN
Erythema nodosum and aseptic arthritis are recognized associations of rickettsial infections. However, they usually present with a febrile illness rather than with severe arthritis. We report three patients who presented with incapacitating major joint arthritis and one who presented with severe spondyloarthropathy in addition to major joint arthritis due to serologically confirmed Orientia tsutsugamushi and Rickettsia conorii infections. All of them had erythema nodosum and low-grade fever. They had rapid clinical response to doxycycline.
Asunto(s)
Artritis/diagnóstico , Eritema Nudoso/diagnóstico , Infecciones por Rickettsia/diagnóstico , Adulto , Antibacterianos/uso terapéutico , Anticuerpos Antibacterianos/sangre , Artritis/tratamiento farmacológico , Artritis/microbiología , Diagnóstico Diferencial , Doxiciclina/uso terapéutico , Eritema Nudoso/tratamiento farmacológico , Eritema Nudoso/microbiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Orientia tsutsugamushi/inmunología , Orientia tsutsugamushi/aislamiento & purificación , Infecciones por Rickettsia/tratamiento farmacológico , Infecciones por Rickettsia/microbiología , Rickettsia conorii/inmunología , Rickettsia conorii/aislamiento & purificación , Resultado del Tratamiento , Adulto JovenAsunto(s)
Interacciones Huésped-Patógeno , Rhipicephalus sanguineus/microbiología , Rickettsia conorii/patogenicidad , Animales , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Chlorocebus aethiops , Técnicas de Cocultivo , ADN Bacteriano/genética , Perros , Larva/microbiología , Reacción en Cadena de la Polimerasa , Análisis de Supervivencia , Células VeroRESUMEN
BACKGROUND: Rickettsial infections are re-emerging. A study of the geographical distribution of rickettsial infections, their clinical manifestations, and their complications would facilitate early diagnosis. METHODS: Thirty-one selected patients from the Western Province of Sri Lanka were studied for rickettsial species, clinical manifestations, and complications. RESULTS: Of 31 patients with possible rickettsioses, 29 (94%) fell into the categories of confirmed, presumptive, or exposed cases of acute rickettsial infections (scrub typhus was diagnosed in 19 (66%), spotted fever group in eight (28%)). Early acute infection or past exposure was suggested in two (7%) cases; cross-reactivity of antigens or past exposure to one or more species was suggested in nine (31%). Seventeen out of 19 (89%) patients with scrub typhus had eschars. Nine out of 29 (32%) patients had a discrete erythematous papular rash: seven caused by spotted fever group, two by scrub typhus. Severe complications were pneumonitis in eight (28%), myocarditis in five (17%), deafness in four (14%), and tinnitus in two (7%). The mean duration of illness before onset of complications was 12.0 (SD 1.4) days. All patients except one made a good clinical recovery with doxycycline or a combination of doxycycline and chloramphenicol. CONCLUSIONS: In a region representing the low country wet zone of Sri Lanka, the main rickettsial agent seems to be Orientia tsutsugamushi. Delay in diagnosis may result in complications. All species responded well to current treatment.
Asunto(s)
Enfermedades Transmisibles Emergentes/epidemiología , Enfermedades Transmisibles Emergentes/microbiología , Infecciones por Rickettsia/epidemiología , Rickettsieae/aislamiento & purificación , Tifus por Ácaros/epidemiología , Adulto , Anciano , Anticuerpos Antibacterianos/aislamiento & purificación , Eritema/epidemiología , Eritema/etiología , Femenino , Hospitales , Humanos , Masculino , Persona de Mediana Edad , Miocarditis/epidemiología , Miocarditis/etiología , Orientia tsutsugamushi/aislamiento & purificación , Neumonía por Rickettsiaceae/epidemiología , Neumonía por Rickettsiaceae/etiología , Infecciones por Rickettsia/sangre , Infecciones por Rickettsia/complicaciones , Tifus por Ácaros/sangre , Tifus por Ácaros/complicaciones , Sri Lanka/epidemiología , Acúfeno/epidemiología , Acúfeno/etiología , Resultado del TratamientoRESUMEN
The human body louse, Pediculus humanus humanus (L.), and the human head louse, Pediculus humanus capitis, belong to the hemimetabolous order Phthiraptera. The body louse is the primary vector that transmits the bacterial agents of louse-borne relapsing fever, trench fever, and epidemic typhus. The genomes of the bacterial causative agents of several of these aforementioned diseases have been sequenced. Thus, determining the body louse genome will enhance studies of host-vector-pathogen interactions. Although not important as a major disease vector, head lice are of major social concern. Resistance to traditional pesticides used to control head and body lice have developed. It is imperative that new molecular targets be discovered for the development of novel compounds to control these insects. No complete genome sequence exists for a hemimetabolous insect species primarily because hemimetabolous insects often have large (2000 Mb) to very large (up to 16,300 Mb) genomes. Fortuitously, we determined that the human body louse has one of the smallest genome sizes known in insects, suggesting it may be a suitable choice as a minimal hemimetabolous genome in which many genes have been eliminated during its adaptation to human parasitism. Because many louse species infest birds and mammals, the body louse genome-sequencing project will facilitate studies of their comparative genomics. A 6-8X coverage of the body louse genome, plus sequenced expressed sequence tags, should provide the entomological, evolutionary biology, medical, and public health communities with useful genetic information.
Asunto(s)
Genoma/genética , Genómica/métodos , Pediculus/genética , Animales , Análisis de Secuencia de ADNRESUMEN
This study describes preliminary results of an investigation of RMSF in Arizona associated with the brown dog tick, Rhipicephalus sanguineus. High numbers of dogs and heavy infestations of ticks created a situation leading to human disease.
Asunto(s)
Rhipicephalus sanguineus/microbiología , Fiebre Maculosa de las Montañas Rocosas/epidemiología , Animales , Arizona/epidemiología , Dermacentor/microbiología , Humanos , IncidenciaRESUMEN
We describe 6 patients with scrub typhus who presented with acute hearing loss, a forgotten complication of this reemerging disease. They were admitted with fever of 10-14 days' duration and had clinical evidence of deafness and pneumonitis. Five patients had eschars, which prompted the diagnosis of typhus fever and led to early institution of treatment. Deafness has been described as a clue to the diagnosis of scrub typhus; awareness of this symptom facilitated early diagnosis in 4 of 5 patients who recovered. Acute hearing loss or hearing impairment in a febrile patient should arouse strong suspicion of scrub typhus.
Asunto(s)
Enfermedades Transmisibles Emergentes/complicaciones , Pérdida Auditiva/microbiología , Tifus por Ácaros/complicaciones , Enfermedad Aguda , Femenino , Humanos , Persona de Mediana EdadRESUMEN
Eleven isolates of spotted fever group rickettsiae from the blood of patients or ixodid ticks from North and South America were characterized. All isolates were identified as Rickettsia rickettsii using restriction fragment length polymorphism analysis of a 532-bp rOmpA gene fragment obtained by PCR. The ability of the R. rickettsii isolates to elicit cytopathic effects and parameters of oxidative injury were examined in cultured human EA.hy 926 endothelial cells. Cytopathic effects were determined by direct observation of infected cultures, by measuring the release of cytoplasmic lactate dehydrogenase (LDH), and by determination of intracellular pools of peroxide and reduced glutathione. Four biotypes of R. rickettsii were defined. Group I included two highly cytopathic isolates from Montana, Bitterroot and Sheila Smith, and three isolates from Maryland, North Carolina, and Brazil. These isolates rapidly damaged cells, released large amounts of cytoplasmic LDH, caused accumulation of intracellular peroxide, and depleted intracellular pools of reduced glutathione. Group II contained three isolates, two from Montana, Hlp#2 and Lost Horse Canyon, and an isolate from Colombia, which were similar to group I but caused either lower responses in LDH release or smaller changes in intracellular peroxide levels. The group III isolates, Sawtooth from Montana and 84JG from North Carolina, caused lower cellular injury by all measures. Group IV isolate Price T from Montana was the least cytopathic and caused minimal alterations of all parameters measured. Understanding the molecular basis for the varied cellular injury caused by different isolates of R. rickettsii may contribute to improved treatment of Rocky Mountain spotted fever and to the rapid identification of those isolates which are more likely to cause fulminant disease.
Asunto(s)
Rickettsia rickettsii/patogenicidad , Animales , Línea Celular , Chlorocebus aethiops , Endotelio/citología , Glutatión/metabolismo , Humanos , Líquido Intracelular/metabolismo , L-Lactato Deshidrogenasa/metabolismo , Peróxidos/metabolismo , Rickettsia rickettsii/aislamiento & purificación , Células VeroRESUMEN
The variable 56-kDa major outer membrane protein of Orientia tsutsugamushi is the immunodominant antigen in human scrub typhus infections. We developed a rapid immunochromatographic flow assay (RFA) for the detection of immunoglobulin M (IgM) and IgG antibodies to O. tsutsugamushi. The RFA employs a truncated recombinant 56-kDa protein from the Karp strain as the antigen. The performance of the RFA was evaluated with a panel of 321 sera (serial bleedings of 85 individuals suspected of scrub typhus) which were collected in the Pescadore Islands, Taiwan, from 1976 to 1977. Among these 85 individuals, IgM tests were negative for 7 cases by both RFA and indirect fluorescence assay (IFA) using Karp whole-cell antigen. In 29 cases specific responses were detected by the RFA earlier than by IFA, 44 cases had the same detection time, and 5 cases were detected earlier by IFA than by RFA. For IgG responses, 4 individuals were negative with both methods, 37 cases exhibited earlier detection by RFA than IFA, 42 cases were detected at the same time, and 2 cases were detected earlier by IFA than by RFA. The sensitivities of RFA detection of antibody in sera from confirmed cases were 74 and 86% for IgM and IgG, respectively. When IgM and IgG results were combined, the sensitivity was 89%. A panel of 78 individual sera collected from patients with no evidence of scrub typhus was used to evaluate the specificity of the RFA. The specificities of the RFA were 99% for IgM and 97% for IgG. The sensitivities of IFA were 53 and 73% for IgM and IgG, respectively, and were 78% when the results of IgM and IgG were combined. The RFA test was significantly better than the IFA test for the early detection of antibody to scrub typhus in primary infections, while both tests were equally sensitive with reinfected individuals.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/inmunología , Inmunoensayo/métodos , Orientia tsutsugamushi/aislamiento & purificación , Tiras Reactivas , Tifus por Ácaros/diagnóstico , Especificidad de Anticuerpos , Antígenos Bacterianos/inmunología , Humanos , Epítopos Inmunodominantes/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Orientia tsutsugamushi/inmunología , Proteínas Recombinantes/inmunología , Sensibilidad y EspecificidadRESUMEN
The genera Anaplasma, Ehrlichia, Cowdria, Neorickettsia and Wolbachia encompass a group of obligate intracellular bacteria that reside in vacuoles of eukaryotic cells and were previously placed in taxa based upon morphological, ecological, epidemiological and clinical characteristics. Recent genetic analyses of 16S rRNA genes, groESL and surface protein genes have indicated that the existing taxa designations are flawed. All 16S rRNA gene and groESL sequences deposited in GenBank prior to 2000 and selected sequences deposited thereafter were aligned and phylogenetic trees and bootstrap values were calculated using the neighbour-joining method and compared with trees generated with maximum-probability, maximum-likelihood, majority-rule consensus and parsimony methods. Supported by bootstrap probabilities of at least 54%, 16S rRNA gene comparisons consistently clustered to yield four distinct clades characterized roughly as Anaplasma (including the Ehrlichia phagocytophila group, Ehrlichia platys and Ehrlichia bovis) with a minimum of 96.1% similarity, Ehrlichia (including Cowdria ruminantium) with a minimum of 97.7% similarity, Wolbachia with a minimum of 95.6% similarity and Neorickettsia (including Ehrlichia sennetsu and Ehrlichia risticii) with a minimum of 94.9% similarity. Maximum similarity between clades ranged from 87.1 to 94.9%. Insufficient differences existed among E. phagocytophila, Ehrlichia equi and the human granulocytic ehrlichiosis (HGE) agent to support separate species designations, and this group was at least 98.2% similar to any Anaplasma species. These 16S rRNA gene analyses are strongly supported by similar groESL clades, as well as biological and antigenic characteristics. It is proposed that all members of the tribes Ehrlichieae and Wolbachieae be transferred to the family Anaplasmataceae and that the tribe structure of the family Rickettsiaceae be eliminated. The genus Anaplasma should be emended to include Anaplasma (Ehrlichia) phagocytophila comb. nov. (which also encompasses the former E. equi and the HGE agent), Anaplasma (Ehrlichia) bovis comb. nov. and Anaplasma (Ehrlichia) platys comb. nov., the genus Ehrlichia should be emended to include Ehrlichia (Cowdria) ruminantium comb. nov. and the genus Neorickettsia should be emended to include Neorickettsia (Ehrlichia) risticii comb. nov. and Neorickettsia (Ehrlichia) sennetsu comb. nov.
Asunto(s)
Anaplasmataceae/clasificación , Proteínas Bacterianas/genética , Chaperoninas/genética , ARN Ribosómico 16S/genética , Rickettsiaceae/clasificación , Anaplasma/clasificación , Anaplasma/genética , Anaplasmataceae/genética , Animales , ADN Ribosómico/genética , Ehrlichia/clasificación , Ehrlichia/genética , Ehrlichia ruminantium/clasificación , Ehrlichia ruminantium/genética , Humanos , Datos de Secuencia Molecular , Filogenia , Rickettsiaceae/genética , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
The 56-kDa major outer membrane protein antigen of Orientia tsutsugamuchi is the immunodominant antigen in human scrub typhus (ST) infections. An enzyme-linked immunosorbent assay (ELISA) using a recombinant 56-kDa protein (r56) to detect specific immunoglobulin M (IgM) produced in ST infections was developed, and its performance was evaluated using sera from patients with active ST (n = 59), spotted fever (SF) (n = 31), and murine typhus (MT) (n = 6) and from those without rickettsial infection (n = 52). The r56 ELISA was compared to an ELISA using native whole cell lysate of O. tsutsugamushi Karp or O. tsutsugamushi Gilliam as antigens. The performance of the assays using r56 was similar to that of those using native antigens. Using indirect immunoperoxidase (IIP) as the reference test, sensitivities were 86, 88, and 88% while specificities were 84, 90, and 87% in the three assays. Furthermore, cross-reactivity in confirmed cases of SF and MT was low (5.4, 2.7, and 2.7% respectively). The additional use of IgG in the r56 ELISA gave improved performance (sensitivity, 80%; specificity, 96%; cross-reactivity in SF and MT, 2.7%). The detection of high levels of IgG in some IgM-negative patients illustrates the importance of including a test for IgG in the detection of secondary or reactivated infections, since many of these patients were from regions in Thailand where these infections are endemic.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Antígenos de Superficie/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Orientia tsutsugamushi/inmunología , Tifus por Ácaros/diagnóstico , Antígenos de Superficie/genética , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/inmunología , Reacciones Cruzadas , Estudios de Evaluación como Asunto , Humanos , Técnicas para Inmunoenzimas , Juego de Reactivos para Diagnóstico , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Infecciones por Rickettsia/diagnóstico , Infecciones por Rickettsia/inmunología , Tifus por Ácaros/inmunología , Sensibilidad y EspecificidadRESUMEN
Heat shock proteins (Hsp) of four Rickettsia species, three Bartonella species, two Ehrlichia species, Orientia tsutsugamushi and seventeen other eubacterial species were characterized by the enhanced chemiluminescence Western blotting (WB) technique with antibodies raised against recombinant Hsp from Escherichia coli and purified GroES from R. typhi. Although E. coli DnaK and GroEL have epitopes that are highly conserved among the homologous proteins found in Rickettsia, Ehrlichia, O. tsutsugamushi, Bartonella and other Proteobacteria, anti-E. coli DnaK and GroEL monoclonal antibodies (Dasch et al. (1990) Ann. N.Y. Acad. Sci. 590, 352-369) recognize less conserved epitopes. In contrast, epitopes on E. coli DnaJ, GrpE and GroES are much less conserved since anti-E. coli DnaJ, GrpE and GroES polyclonal antibodies did not recognize DnaJ, GrpE or GroES homologues in Rickettsia, Bartonella, Orientia, Ehrlichia and Legionella. Polyclonal antiserum prepared against GroES from R. typhi reacted strongly with purified 10 kDa GroES peptide from Rickettsia and Bartonella, and strongly bound to proteins of varying electrophoretic mobility from Wolbachia, Legionella, Proteus and Shigella flexneri and more weakly to other GroES homologues including that found in E. coli. Consequently, commercially available anti-DnaJ, anti-GrpE and anti-GroES polyclonal antibodies and anti-DnaK monoclonal antibody raised against their respective recombinant E. coli Hsp are not suitable for detection and identification of homologues of these proteins in a wide range of eubacteria.
Asunto(s)
Alphaproteobacteria/química , Bacterias/química , Western Blotting , Proteínas de Choque Térmico/química , Alphaproteobacteria/crecimiento & desarrollo , Animales , Anticuerpos Antibacterianos/inmunología , Anticuerpos Monoclonales/inmunología , Bacterias/crecimiento & desarrollo , Reacciones Cruzadas , Electroforesis en Gel de Poliacrilamida , Proteínas de Choque Térmico/inmunología , Respuesta al Choque Térmico , Ratones , ConejosRESUMEN
The variable 56-kDa major outer membrane protein of Orientia tsutsugamushi is the immunodominant antigen in human scrub typhus infections. The gene encoding this protein from Karp strain was cloned into the expression vector pET11a. The recombinant protein (r56) was expressed as a truncated nonfusion protein (amino acids 80 to 456 of the open reading frame) which formed an inclusion body when expressed in Escherichia coli BL21. Refolded r56 was purified and compared to purified whole-cell lysate of the Karp strain of O. tsutsugamushi by immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) for reactivity with rabbit sera prepared against eight antigenic prototypes of O. tsutsugamushi as well as several other species of Rickettsiales and nonrickettsial antigens. Refolded r56 exhibited broad reactivity with the rabbit antisera against the Orientia prototypes, and the ELISA reactions with the r56 and Karp whole-cell lysate antigens correlated well (r = 0.81, n = 22, sensitivity compared to that of standard ELISA of 91%). Refolded r56 did not react with most antisera against other rickettsial species or control antigens (specificity = 92%, n = 13) using a positive cutoff value determined with eight uninfected rabbit sera. Refolded r56 was evaluated further by ELISA, using 128 sera obtained from patients with suspected scrub typhus from Korat, Thailand, and 74 serum specimens from healthy Thai soldiers. By using the indirect immunoperoxidase assay as the reference assay, the recombinant antigen exhibited a sensitivity and specificity of 93% or greater for detection of both IgG and IgM in the ELISA at 1:400 serum dilution. These results strongly suggest that purified r56 is a suitable candidate for replacing the density gradient-purified, rickettsia-derived, whole-cell antigen currently used in the commercial dipstick assay available in the United States.
Asunto(s)
Antígenos Bacterianos/química , Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Orientia tsutsugamushi/genética , Orientia tsutsugamushi/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Antibacterianos/sangre , Antígenos Bacterianos/aislamiento & purificación , Proteínas de la Membrana Bacteriana Externa/química , Secuencia de Bases , Estudios de Casos y Controles , Cartilla de ADN/genética , Ensayo de Inmunoadsorción Enzimática/estadística & datos numéricos , Expresión Génica , Genes Bacterianos , Humanos , Técnicas para Inmunoenzimas/estadística & datos numéricos , Datos de Secuencia Molecular , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/inmunología , Pliegue de Proteína , Conejos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Tifus por Ácaros/diagnóstico , Tifus por Ácaros/inmunología , Sensibilidad y EspecificidadRESUMEN
The cytoplasmic 120 kDa antigen genes of 9 isolates of Rickettsia conorii (RC), 12 isolates of R. africae (RA), and 3 isolates of Israeli tick typhus rickettsiae (ISTT) were compared for restriction fragment length polymorphisms (RFLP) present in portions of the open reading frame amplified by polymerase chain reaction (PCR). Initially, DNAs from 13 species or serotypes of spotted fever group rickettsiae were used to select restriction enzymes (RE) that detected RFLP in gene fragments amplified with primer pairs 483WF/1514R and 764F/3409R. Among the R. conorii complex isolates, Dpn II gave RFLP differentiating all three serotypes. Unique RE patterns were obtained for RC with Bsr I and Hinf I, for RA with Mwo I, Pst I and Ssp I, and for ISTT with Hpa II. While RFLP typing of the 120 kDa gene permitted rapid separation of R. conorii complex isolates into three groups corresponding to the RC, RA, and ISTT rOmp serotypes, additional intragroup genetic variation was also detected in all three serotypes.
Asunto(s)
Polimorfismo de Longitud del Fragmento de Restricción , Infecciones por Rickettsia/microbiología , Rickettsia/genética , Animales , Secuencia de Bases , Chlorocebus aethiops , Cartilla de ADN , ADN Bacteriano/aislamiento & purificación , Variación Genética , Humanos , Israel , Células L , Ratones , Reacción en Cadena de la Polimerasa , Mapeo Restrictivo , Rickettsia/clasificación , Rickettsia/aislamiento & purificación , Células VeroRESUMEN
BACKGROUND: The purpose of this study was to determine whether infective Orientia tsutsugamushi, the etiologic agent of scrub typhus, could survive normal blood banking processing and storage procedures. STUDY DESIGN AND METHODS: Mononuclear cells isolated from whole blood by density gradient centrifugation were inoculated with O. tsutsugamushi, Karp strain. Infection of the mononuclear cells was confirmed by Giemsa stain, direct fluorescent antibody assay, and polymerase chain reaction using primers specific for the groESL operon of O. tsutsugamushi. The quantity of rickettsial particles in each preparation was determined by direct counts from the Giemsa-stained preparations. Infected mononuclear cells were returned to their respective aliquots of packed red blood cells, which were then either stored at 4 degrees C or glycerolized and frozen at -70 degrees C. RESULTS: Rickettsiae survived up to 10 days (but not 30 days) of refrigerated storage and 45 days of frozen storage, as determined by inoculation of mice with 0.5-mL aliquots of the blood components. Infection of the mice was determined by illness, death, direct fluorescent antibody assay of peritoneal smears, polymerase chain reaction of blood, and enzyme-linked immunosorbent assay detection of antibodies in plasma. CONCLUSION: Because the quantity of rickettsiae injected into the mice was comparable to the quantity reported in the literature for human blood during natural infections, scrub typhus could present a risk in blood collected from donors in endemic areas. This may especially be true, because people can be rickettsemic before illness, after successful antibiotic treatment, and chronically after resolution of disease.
Asunto(s)
Transfusión de Eritrocitos/efectos adversos , Eritrocitos/microbiología , Orientia tsutsugamushi/aislamiento & purificación , Tifus por Ácaros/transmisión , Animales , Bancos de Sangre/normas , Humanos , Ratones , Orientia tsutsugamushi/fisiologíaRESUMEN
The lipopolysaccharides (LPSs) isolated from typhus group (TG) rickettsiae Rickettsia typhi and Rickettsia prowazekii were characterized by chemical analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining. LPSs from two species of TG rickettsiae contained glucose, 3-deoxy-D-manno-octulosonic acid, glucosamine, quinovosamine, phosphate, and fatty acids (beta-hydroxylmyristic acid and heneicosanoic acid) but not heptose. The O-polysaccharides of these LPSs were composed of glucose, glucosamine, quinovosamine, and phosphorylated hexosamine. Resolution of these LPSs by their apparent molecular masses by SDS-PAGE showed that they have a common ladder-like pattern. Based on the results of chemical composition and SDS-PAGE pattern, we suggest that these LPSs act as group-specific antigens. Furthermore, glucosamine, quinovosamine, and phosphorylated hexosamine were also found in the O-polysaccharide of the LPS from Proteus vulgaris OX19 used in the Weil-Felix test, suggesting that they may represent the antigens common to LPSs from TG rickettsiae and P. vulgaris OX19.