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1.
Clin Infect Dis ; 29(2): 426-8, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10476753

RESUMO

The clinical characteristics of 26 patients with community-acquired pneumonia due to Chlamydia pneumoniae as the only identified pathogen who required hospitalization were evaluated. Most patients (18) had reinfection based on serological results. The mean age of the patients was 55 years (38 years, patients with primary infection; 63 years, patients with reinfection), and the gender representation was equal. Generally, illness was mild and associated with limited temperature elevation and nonspecific symptoms. The presence of comorbid illnesses and the requirement of supplemental oxygen therapy were the most common criteria for hospital admission.


Assuntos
Infecções por Chlamydia/microbiologia , Infecções por Chlamydia/fisiopatologia , Chlamydophila pneumoniae , Infecções Comunitárias Adquiridas/microbiologia , Pneumonia/fisiopatologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antibacterianos/sangue , Infecções por Chlamydia/sangue , Infecções por Chlamydia/imunologia , Chlamydophila pneumoniae/imunologia , Infecções Comunitárias Adquiridas/sangue , Infecções Comunitárias Adquiridas/imunologia , Infecções Comunitárias Adquiridas/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pneumonia/sangue , Pneumonia/imunologia , Pneumonia/microbiologia
2.
Clin Infect Dis ; 26(6): 1425-9, 1998 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9636874

RESUMO

Avian chlamydiosis was detected in a shipment of > 700 pet birds from a Florida bird distributor that were sold to nine Atlanta-area pet stores in August 1995. Respiratory illness among persons who had recently acquired birds from this shipment was reported to local public health officials. The attack rate of acute respiratory illness was 10.7% among persons in households exposed to birds from the implicated flock vs. 1.8% among control households (odds ratio, 6.60; 95% confidence interval, 1.39-31.2). Illness and serological evidence of infection in the absence of symptoms were more common among persons in households with recently purchased birds that were sick or that had died and among persons who had had direct contact with the birds. Clinical psittacosis or serological evidence of Chlamydia psittaci infection was found in 30.7% of households with birds from the infected flock. Mild illnesses and asymptomatic infections in exposed persons were unusual features of this outbreak.


Assuntos
Doenças das Aves/parasitologia , Aves/parasitologia , Chlamydophila psittaci , Surtos de Doenças , Psitacose/etiologia , Zoonoses/parasitologia , Animais , Georgia , Humanos
3.
J Clin Lab Anal ; 12(1): 47-53, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9484669

RESUMO

We evaluated multiple procedures for extracting chlamydial DNA from specimens for detection in PCR tests. Commercial kits and an in-house method were tested for their sensitivity and utility. Quantifiable chlamydial elementary bodies (EB) were used for spiking buffy coats from EDTA-collected blood. EBs of Chlamydia pneumoniae at 2,500 and 25 EB/ml were used as specimens for DNA extraction using seven different procedures. These included either columns (3 procedures), centrifugation (1), glass (1), or patented extraction matrices (2), coupled with either alcohol precipitation (6) or heat-detergent treatment (1). Five procedures required 10-40 minutes manipulation; two required 2-5 hours. PCR results for DNA extracts using chlamydial 16S genus primers were generally more intensely positive with denser bands on electrophoresis gels for the higher concentrations of EB (up to 4+ for stained product on gels) than was PCR with lower EB concentrations (up to 2+). Further, the incidence of procedures with positive results was: 5 of 7 for chlamydial genus primers with 5 EB vs. 6 of 7 with 500 EB. Maximal sensitivity for one of the extractions was in the range of 2.5-5.0 EB/ml of test specimen with 4 of 5 replicates being positive with EB controls or extracts. Extracts were stable up to 2+ weeks at 4 degrees C and were effective in multiplexing with fluorescent-tagged primers. Taking into consideration the time factor and sensitivities, the two procedures with extraction matrices are favored for routine laboratory use.


Assuntos
Chlamydia/genética , DNA Bacteriano/isolamento & purificação , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade
4.
J Clin Microbiol ; 35(8): 2043-6, 1997 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9230378

RESUMO

We developed a nested, multiplex PCR for simultaneous detection of three species of chlamydiae in human and avian specimens. The PCR was designed to increase sensitivity and to circumvent inhibitors of PCR present in clinical specimens. The target sequence was the 16S rRNA gene. The first-step PCR was genus specific, and the second-step PCR was multiplexed (i.e., had multiple primer sets in the same tube) and could discriminate among Chlamydia pneumoniae, Chlamydia psittaci, and Chlamydia trachomatis on the basis of the molecular weight of the amplicon. The limit of detection of each of the two PCR steps was 5 inclusion-forming units. We used PCR and serologic evidence during outbreaks of psittacosis to infer that C. psittaci had been transmitted from birds purchased in pet stores to humans. We also used this method to test both live and dead birds from pet stores for infection with C. psittaci. Compared with culture, the application of PCR to avian specimens increased the rate of C. psittaci detection.


Assuntos
Chlamydia/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Psitacose/genética , RNA Bacteriano/análise , RNA Ribossômico 16S/análise , Animais , Aves , Chlamydia/classificação , Chlamydia/genética , Chlamydia trachomatis/classificação , Chlamydia trachomatis/isolamento & purificação , Chlamydophila pneumoniae/classificação , Chlamydophila pneumoniae/isolamento & purificação , Chlamydophila psittaci/classificação , Chlamydophila psittaci/isolamento & purificação , Fezes/microbiologia , Psitacose/diagnóstico , Psitacose/epidemiologia
5.
J Clin Microbiol ; 32(10): 2417-21, 1994 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7814477

RESUMO

The serodiagnosis of human psittacosis was considerably improved by a microimmunofluorescence (MIF) assay that uses selected strains of Chlamydia psittaci, C. pneumoniae, and C. trachomatis as antigens. The 78 patients examined in the study were clinically diagnosed as having psittacosis on the basis of compatible clinical symptoms following exposure to sick birds. The conventional complement fixation (CF) test identified 36 patients, or 46% (36 of 78) of the total, as positive. Antibody responses to C. psittaci were demonstrated by the MIF test in all 36 CF-positive patients. The MIF test also detected antibody responses to C. psittaci in 12 patients (15% of the total) whose sera were negative or anticomplementary in the CF test. Seven patients, or 9% (7 of 78) of the total, were identified by the MIF test as having C. pneumoniae infections. About 30% of the study patients (23 of 78) showed no serologic evidence of either C. psittaci or C. pneumoniae infection by both the CF and the MIF tests. Four distinctive serologic reaction patterns were observed in the study patients. Recognition of these reaction patterns and judicious corroboration of serologic responses to the chlamydial species by the MIF test with epidemiologic and clinical information will increase the efficiency and accuracy of serodiagnosis for human psittacosis.


Assuntos
Anticorpos Antibacterianos/sangue , Chlamydophila psittaci/imunologia , Psitacose/diagnóstico , Adolescente , Adulto , Idoso , Criança , Testes de Fixação de Complemento , Imunofluorescência , Humanos , Pessoa de Meia-Idade
6.
Radiology ; 192(3): 819-24, 1994 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8058954

RESUMO

PURPOSE: To report the spectrum of radiographic findings associated with a new respiratory pathogen: Chlamydia pneumoniae (TWAR strain). MATERIALS AND METHODS: Radiographs of 55 adult patients hospitalized with serologic evidence of C pneumoniae were retrospectively reviewed. RESULTS: On the basis of serologic criteria, two types of acute respiratory infection are possible: primary (first exposure) infections and recurrent, acute infection in a previously exposed individual. In the primary group, alveolar opacities (65%) with a unilateral distribution (71%) were most common at admission. Cavitary disease and hilar or mediastinal lymphadenopathy were uncommon. Small to medium-sized pleural effusions were common in both primary and recurrent groups during hospitalization. Also, both groups tended to progress to bilateral, mixed, interstitial and alveolar changes during the course of infection. CONCLUSION: Different radiographic patterns exist for the two types of acute C pneumoniae infection.


Assuntos
Infecções por Chlamydia/diagnóstico por imagem , Chlamydophila pneumoniae , Infecções Respiratórias/diagnóstico por imagem , Doença Aguda , Chlamydophila pneumoniae/classificação , Feminino , Humanos , Pulmão/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Radiografia , Recidiva , Estudos Retrospectivos
7.
J Clin Microbiol ; 30(7): 1625-30, 1992 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-1629316

RESUMO

Two established cell lines, H 292 and HEp-2, originating from the human respiratory tract, were found to be significantly more efficient and practical than the currently used HeLa 229 cells for growth of Chlamydia pneumoniae. Six strains of C. pneumoniae recently isolated from patients with respiratory ailments were used as test cultures. The H 292 and HEp-2 cells yielded much higher inclusion counts for all the test strains than did HeLa 229 cells. When they were compared with each other, H 292 cells yielded more inclusions than did HEp-2 cells, and the differences were statistically significant in 10 of 18 test sets. A simple system with these two cell lines appeared to be very efficient for culturing C. pneumoniae. It does not require treatment of tissue cells with DEAE-dextran before infection, and it may eliminate the need for serial subpassages of specimens to increase culture sensitivity. Monolayers of these cells remained intact and viable in the Chlamydia growth medium so that reinfection could take place, resulting in greatly increased inclusion counts for specimens containing few infectious units. This system may make it more practical for laboratories to culture for C. pneumoniae for treatment of infections and outbreak intervention and will facilitate studies on this recently recognized pathogen.


Assuntos
Linhagem Celular/microbiologia , Chlamydophila pneumoniae/crescimento & desenvolvimento , Anticorpos Monoclonais , DEAE-Dextrano/farmacologia , Humanos , Sistema Respiratório/microbiologia
8.
J Clin Microbiol ; 28(5): 1062-5, 1990 May.
Artigo em Inglês | MEDLINE | ID: mdl-2351723

RESUMO

Two major antigens of Bordetella pertussis, filamentous hemagglutinin (FHA) and pertussis toxin (PT), were efficiently purified from culture filtrate by exploiting their relative hydrophobicities and differences in affinity to sialic acid-containing protein. High yields of FHA (40 to 80 mg/liter) and PT (8 to 16 mg/liter) were first produced by growing the bacteria in the modified CL medium. The FHA and PT in the culture filtrate were adsorbed onto butyl-Sepharose by hydrophobic interaction at appropriately high ionic strength. Elution of the antigens was effected by decreasing their hydrophobicities with a buffer of low ionic strength. FHA was then separated from PT with an affinity column of fetuin-Sepharose. The fraction passing through the column contained purified FHA, and the fetuin-bound PT was eluted with buffered MgCl2. The FHA and PT purified by these steps were electrophoretically and serologically identical to the reference purified FHA and PT preparations. Approximately 16 to 32 mg of purified FHA and 4 to 8 mg of purified PT were obtained from 1 liter of culture filtrate. The described procedure for making FHA and PT antigens from B. pertussis for serologic and immunologic use is very simple, efficient, and reproducible.


Assuntos
Bordetella pertussis/análise , Hemaglutininas/isolamento & purificação , Toxina Pertussis , Fatores de Virulência de Bordetella/isolamento & purificação , Antígenos de Bactérias/isolamento & purificação , Bordetella pertussis/imunologia , Cromatografia de Afinidade , Concentração Osmolar
9.
J Clin Microbiol ; 27(12): 2805-10, 1989 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-2556434

RESUMO

A procedure that is sufficiently simple and economical for use in clinical and public health laboratories for producing and purifying filamentous hemagglutinin (FHA) and determining antibodies to this major antigen of Bordetella pertussis in serum is described. High yields of FHA (40 to 80 mg/liter) were obtained in the supernatant by cultivating B. pertussis in modified CL medium. The FHA antigen was separated from pertussis toxin (PT) and other antigens by chromatography on hydroxylapatite. Removal of residual PT activity in the FHA fraction was effected by affinity absorption of PT with Fetuin immobilized to Sepharose 4B. The FHA was used as the antigen for determining titers of immunoglobulin G (IgG), IgA, and IgM to FHA in sera of patients with pertussis by an improved enzyme-linked immunosorbent assay. Development of the interfering background color commonly observed in conventional FHA enzyme-linked immunosorbent assay procedures was eliminated by washing the reaction wells with a buffer of high ionic strength before adding the peroxidase conjugates. In the absence of nonspecific background color, the reaction endpoints were easy to read. The FHA prepared by the procedure described was identical to a reference preparation of purified FHA in sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles and serological specificity assays. High yields of FHA were obtained from all four strains of B. pertussis tested in this study, indicating that the procedure for enhanced production of FHA may be generally applicable to other strains of B. pertussis. Results from tests of 50 serum specimens with clinical information on pertussis for FHA and PT antibodies by the assay procedures described exemplified the usefulness and caveats of serodiagnosis for pertussis.


Assuntos
Anticorpos Antibacterianos/análise , Bordetella pertussis/imunologia , Hemaglutininas/isolamento & purificação , Toxina Pertussis , Fatores de Virulência de Bordetella/imunologia , Coqueluche/diagnóstico , Bordetella pertussis/crescimento & desenvolvimento , Meios de Cultura , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Hemaglutininas/imunologia , Humanos , Imunoglobulina A/análise , Imunoglobulina G/análise , Imunoglobulina M/análise
10.
J Clin Microbiol ; 26(7): 1316-20, 1988 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-2900845

RESUMO

A new, practical method for determining antibody to pertussis toxin (PT) by enzyme-linked immunosorbent assay for public health and clinical laboratories is possible because of recent advances in understanding the pathobiology of Bordetella pertussis and the physicochemical properties of PT. The new approach does not require the use of highly purified PT antigen, which is difficult and expensive for most laboratories to obtain. Moreover, it employs only reagents that are commercially readily available. The method combines the purification of PT antigen and an assay for PT antibody into one process. It depends on (i) growth of B. pertussis in a simple, defined medium to obtain a PT-rich supernatant with little contamination of cellular antigens; (ii) a simple, one-step concentration of PT in the culture supernatant with Affi-Gel Blue (Bio-Rad Laboratories, Richmond, Calif.); and (iii) specific adsorption of PT as the test antigen to microtiter wells coated with fetuin for the enzyme-linked immunosorbent assay. The procedure is sensitive and specific for PT antibody. It is technically simple, reproducible, and can be performed in a modestly equipped laboratory.


Assuntos
Anticorpos Antibacterianos/análise , Toxina Pertussis , Fatores de Virulência de Bordetella/imunologia , Animais , Antígenos de Bactérias/isolamento & purificação , Bordetella pertussis/crescimento & desenvolvimento , Bordetella pertussis/imunologia , Cromatografia de Afinidade , Meios de Cultura , Ensaio de Imunoadsorção Enzimática , Humanos , Soros Imunes/imunologia , Imunoglobulina A/análise , Imunoglobulina G/análise , Imunoglobulina M/análise , Valor Preditivo dos Testes , Coelhos
11.
J Clin Microbiol ; 23(3): 407-10, 1986 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-3754264

RESUMO

Strains of Campylobacter jejuni and Campylobacter coli were characterized and grouped by their distinct reaction patterns with lectins. Heating of the Campylobacter cultures to 100 degrees C and holding for 30 to 60 min greatly enhanced their reactivity with lectins and permitted the grouping of all but 3 of 155 cultures tested in this study without interference of autoagglutination and other nonspecific activities. The lectin reaction patterns of the heated cultures were stable and reproducible. They were strain specific and independent of the heat-stable antigenic types. The lectin-reactive sites of C. jejuni and C. coli may be useful as additional markers for strain characterization. Based on these observations, a simple slide agglutination procedure is described for differentiating strains of C. jejuni and C. coli by their interaction with a selected group of commercially available lectins.


Assuntos
Campylobacter fetus/classificação , Campylobacter/classificação , Lectinas/farmacologia , Lectinas de Plantas , Aglutinação , Testes de Aglutinação , Campylobacter/metabolismo , Campylobacter fetus/metabolismo , Temperatura Alta , Aglutinina de Amendoim , Receptores de N-Acetilglucosamina , Aglutininas do Germe de Trigo
12.
J Clin Microbiol ; 22(1): 134-5, 1985 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-2410445

RESUMO

Lectins and blood group antibodies were used to probe the surface structures of Campylobacter jejuni and Campylobacter coli. Of the 29 strains tested, there were distinct reaction patterns. The lectin-reactive and blood group antibody-reactive sites on the bacterial surface were distinguishable from the heat-stable (lipopolysaccharide) antigenic determinants. The interactions were strain specific. The reactive sites were stable with respect to culture media and passage and may be useful as additional markers for strain characterization.


Assuntos
Antígenos de Bactérias/análise , Antígenos de Superfície/análise , Antígenos de Grupos Sanguíneos/imunologia , Campylobacter fetus/imunologia , Campylobacter/imunologia , Lectinas , Testes de Aglutinação , Campylobacter/classificação , Campylobacter fetus/classificação , Epitopos , Lectinas/imunologia , Lipopolissacarídeos , Fito-Hemaglutininas , Proteínas de Plantas , Especificidade da Espécie
13.
J Clin Microbiol ; 21(5): 702-7, 1985 May.
Artigo em Inglês | MEDLINE | ID: mdl-3998098

RESUMO

A coagglutination system has been devised for typing heat-stable and heat-labile antigens of Campylobacter jejuni and C. coli. The use of protein A-positive Staphylococcus aureus cells carrying Campylobacter sp. serotype antibody and the treatment of Campylobacter sp. cells with DNase in the antigen suspension permitted rapid and specific coagglutination of rough (autoagglutinable) as well as smooth cultures. Cells of S. aureus were sensitized with Campylobacter sp. serotype antisera. Four to five types of sensitized S. aureus cells were pooled. A strain of Campylobacter sp. was first tested with the pools and then typed with the individual reagents of the reactive pool. After the described procedures, 68 serotype strains tested blindly as unknowns were correctly typed according to their heat-stable or heat-labile antigens. The two most commonly used typing schemes which are based separately on the heat-stable or the heat-labile antigens as assayed by passive hemagglutination and slide agglutination, respectively, can be utilized simultaneously in the coagglutination system for strain characterization. The coagglutination system is simple, yields results rapidly, conserves typing reagents, and offers the flexibility of formulating the pools of reagents according to the experimental design or the prevalence of serotypes in a geographic location. It should be a practical system for the typing of Campylobacter spp. in public health or clinical laboratories.


Assuntos
Antígenos de Bactérias/análise , Campylobacter fetus/imunologia , Campylobacter/imunologia , Testes de Aglutinação , Animais , Campylobacter/classificação , Temperatura Alta , Coelhos , Sorotipagem
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