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1.
Sex Transm Infect ; 81(6): 463-6, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16326847

RESUMO

OBJECTIVES: Mycoplasma genitalium has been associated with cervicitis, endometritis, and tubal factor infertility. Because the ability of this bacterium to ascend and infect the fallopian tube remains undefined, we performed an investigation to determine the prevalence of M genitalium in fallopian tube, endometrial, and cervical specimens from women laparoscopically diagnosed with acute salpingitis in Nairobi, Kenya. METHODS: Women presenting with pelvic inflammatory disease were laparoscopically diagnosed with salpingitis. Infection with M genitalium in genital specimens was determined by polymerase chain reaction (PCR). RESULTS: Of 123 subjects with acute salpingitis, M genitalium was detected by PCR in the cervix and/or endometrium in nine (7%) participants, and in a single fallopian tube specimen. In addition, those infected with M genitalium were more often HIV infected than women not infected by M genitalium (seven of nine (78%) v 42 of 114 (37%), p<0.03). CONCLUSIONS: M genitalium is able to ascend into the fallopian tube, but its association with tubal pathology requires further investigation.


Assuntos
Laparoscopia/métodos , Infecções por Mycoplasma/diagnóstico , Mycoplasma genitalium/isolamento & purificação , Salpingite/diagnóstico , Doença Aguda , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Reação em Cadeia da Polimerase/métodos , Estudos Prospectivos , Salpingite/microbiologia
2.
Am J Med ; 108(4): 290-5, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11014721

RESUMO

PURPOSE: To describe a nosocomial outbreak of Legionella micdadei pneumonia in transplant patients and to characterize the source of the outbreak and the control measures utilized. SUBJECTS AND METHODS: We performed retrospective Legionella micdadei serologic testing to enhance case finding in transplant patients with pneumonia that lacked a documented microbial etiology, as well as prospective environmental surveillance of water sites and testing for Legionella in clinical specimens. RESULTS: During a 3-month period, 12 cases of Legionella micdadei pneumonia were identified either by culture or serologic testing among 38 renal and cardiac transplant patients. Legionella micdadei isolates from hot water sources were found by pulsed-field gel electrophoresis to have a DNA banding pattern that was identical to the isolates from the first 3 culture-positive cases and from 2 cases that occurred 16 months later. CONCLUSIONS: Hospitals caring for organ transplant recipients and other immunosuppressed patients must be aware of the possibility of environmental sources of outbreaks of Legionella infection. A first-line screen with the Legionella urine antigen test will identify Legionella pneumophila serogroup 1. However, specific cultures in outbreak situations should be considered to identify other Legionella pneumophila serotypes and the nonpneumophila Legionella species.


Assuntos
Surtos de Doenças , Transplante de Coração , Controle de Infecções/métodos , Transplante de Rim , Legionella/isolamento & purificação , Doença dos Legionários/epidemiologia , Complicações Pós-Operatórias/microbiologia , Adulto , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , Surtos de Doenças/prevenção & controle , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Legionella/genética , Doença dos Legionários/microbiologia , Doença dos Legionários/prevenção & controle , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Cidade de Nova Iorque/epidemiologia , Complicações Pós-Operatórias/epidemiologia , Estudos Retrospectivos
3.
J Clin Microbiol ; 38(9): 3341-8, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10970381

RESUMO

Accurate antimicrobial susceptibility testing is vital for patient care and surveillance of emerging antimicrobial resistance. The National Committee for Clinical Laboratory Standards (NCCLS) outlines generally agreed upon guidelines for reliable and reproducible results. In January 1997 we surveyed 320 laboratories participating in the New York State Clinical Evaluation Program for General Bacteriology proficiency testing. Our survey addressed compliance with NCCLS susceptibility testing guidelines for bacterial species designated a problem (Staphylococcus aureus and Enterococcus species) or fastidious (Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria gonorrhoeae) organism. Specifically, we assessed compliance with guidelines for inoculum preparation, medium choice, number of disks per plate, and incubation conditions for disk diffusion tests. We also included length of incubation for S. aureus and Enterococcus species. We found overall compliance with the five characteristics listed above in 80 of 153 responding laboratories (50.6%) for S. aureus and 72 of 151 (47.7%) laboratories for Enterococcus species. The most common problem was an incubation time shortened to less than 24 h. Overall compliance with the first four characteristics was reported by 92 of 221 (41.6%) laboratories for S. pneumoniae, 49 of 163 (30.1%) laboratories for H. influenzae, and 11 of 77 (14.3%) laboratories for N. gonorrhoeae. Laboratories varied from NCCLS guidelines by placing an excess number of disks per plate. Laboratories also reported using alternative media for Enterococcus species, N. gonorrhoeae, and H. influenzae. This study demonstrates a need for education among clinical laboratories to increase compliance with NCCLS guidelines.


Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Infecções Bacterianas/microbiologia , Laboratórios/normas , Testes de Sensibilidade Microbiana/normas , Guias como Assunto , Humanos , Testes de Sensibilidade Microbiana/métodos , New York , Reprodutibilidade dos Testes
4.
J Vet Diagn Invest ; 8(2): 186-95, 1996 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8744740

RESUMO

Aerotolerant organisms resembling Campylobacter, now designated as Arcobacter, have been described from aborted farm animals and from cases of human enteritis worldwide. The goals of this study were 1) to attempt to recover Arcobacter spp. from cases of porcine abortion, 2) to characterize these isolates by phenotype and ribotype, and 3) to compare the usefulness of ribotype and phenotype patterns for identifying Arcobacter butzleri and the DNA hybridization groups 1A and 1B of A. cryaerophilus. Isolates of Arcobacter spp. from North Carolina and Iowa were recovered from porcine tissues. In Iowa, Arcobacter spp. were recovered from 43% (13/30) of porcine abortion cases evaluated. Isolations were made from placenta (44%), kidney (44%), and stomach contents (12%), which were the only tissues examined. The most reliable biochemical tests for A. butzleri included growth in 1% glycine and in 1.5% NaCl, weak catalase activity, and resistance to cadmium chloride. Arcobacter cryaerophilus strains were characterized by strong catalase activity and sensitivity to cadmium chloride. The DNA hybridization groups 1A and 1B of A. cryaerophilus could not be distinguished by biochemical tests. This represents the first description of A. cryaerophilus DNA group 1A in animals within the United States.


Assuntos
Aborto Animal/microbiologia , Infecções por Campylobacter/veterinária , Campylobacter/classificação , Campylobacter/isolamento & purificação , Enterite/veterinária , Doenças dos Suínos , Animais , Campylobacter/genética , Infecções por Campylobacter/diagnóstico , Cromossomos Bacterianos , Enterite/diagnóstico , Feminino , Humanos , Fenótipo , Placenta/microbiologia , Gravidez , RNA Ribossômico/genética , RNA Ribossômico/isolamento & purificação , Suínos
5.
J Clin Microbiol ; 33(11): 2940-7, 1995 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8576350

RESUMO

By DNA-DNA hybridization, we classified 26 human strains, 4 dog and cat strains, and 4 hamster strains putatively identified as Helicobacter cinaedi as well as 2 human strains and 2 animal strains of Helicobacter fennelliae. All but one human strain belonged to the same hybridization group as the type strain of H. cinaedi. The animal strains also appeared to belong to this hybridization group. Both human strains of H. fennelliae were shown to be H. fennelliae by DNA-DNA hybridization, but both animal strains were less than 15% related to the type strain. All strains were also characterized by plasmid profiles and ribotyping. Plasmids were found in 23% of the human strains, 100% of the hamster strains, and 33% of the dog and cat strains. Human strains were essentially identical by ribotyping, but were clearly differentiated from the hamster and dog and cat strains. Some strains may be difficult to culture on primary isolation; we found that our strains grew well on anaerobic CDC agar, brucella agar, and tryptic soy agar II. Our H. cinaedi and H. fennelliae strains differed from those previously described because some were resistant to cephalothin: some H. cinaedi strains were also resistant to nalidixic acid. All isolates were also characterized by antimicrobial susceptibility testing. We found that human strains of H. cinaedi were more resistant to clindamycin and erythromycin than were animal isolates; 19% of the human strains were resistant to ciprofloxacin. Therefore, we recommend that antimicrobial susceptibility results be obtained before initiating therapy for H. cinaedi and H. fennelliae infections.


Assuntos
Técnicas de Tipagem Bacteriana , Helicobacter/classificação , Animais , Gatos , Cricetinae , Sondas de DNA , DNA Bacteriano , DNA Ribossômico , Cães , Genótipo , Helicobacter/genética , Helicobacter/crescimento & desenvolvimento , Helicobacter/metabolismo , Infecções por Helicobacter/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Fenótipo , Plasmídeos , Especificidade da Espécie
6.
Lab Anim Sci ; 44(6): 579-83, 1994 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-7534848

RESUMO

Heat-stable (HS, O-antigen) and heat-labile (HL) serotyping are the most common methods used to type Campylobacter jejuni and C. coli for epidemiologic purposes. In this study, we conducted RRNA analysis to differentiate strains of C. jejuni and C. coli that had been serotyped by use of the passive hemagglutination (heat-stable) and slide agglutination (heat-labile) methods. Ribotyping of isolates within HS and HL serotypes revealed further discrimination of strains. Four ribotypes were identified by Pvu II and Pst I digests of eight HS serotype-34 isolates. Ribotyping also differentiated strains within HL serotypes. Ribotyping also was conducted on 10 representative isolates of C. jejuni and C. coli isolated from an infant macaque. The eight ribotypes confirmed previous results of serotyping and other phenotypic analyses, which indicated that the infant was repeatedly reinfected with different strains of C. jejuni and C. coli. Results of the study indicated that ribotyping is a sensitive molecular marker for distinguishing strains of C. jejuni and C. coli. Furthermore, some isolates with similar ribotype patterns had variability in their HS and HL serotypes.


Assuntos
Campylobacter coli/genética , Campylobacter jejuni/genética , Macaca nemestrina/microbiologia , RNA Bacteriano/análise , RNA Ribossômico/análise , Animais , Campylobacter coli/classificação , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/classificação , Campylobacter jejuni/isolamento & purificação , Desoxirribonucleases de Sítio Específico do Tipo II , Temperatura Alta , Antígenos O , Polissacarídeos Bacterianos/análise , Sorotipagem
7.
Ann Intern Med ; 121(2): 90-3, 1994 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-8017741

RESUMO

OBJECTIVE: To define the clinical spectrum of illness associated with Helicobacter cinaedi infection in the United States and to determine associated epidemiologic risk factors and optimal laboratory methods for recovery of H. cinaedi. DESIGN: A retrospective epidemiologic study of 23 patients with H. cinaedi-associated illness. PATIENTS: 23 patients with H. cinaedi infection identified between January 1982 and August 1990. Most isolates (22 of 23) were from blood; one was from stool. RESULTS: Ages ranged from 24 to 84 years (mean, 44 years). Eighty-three percent of patients were men; 17% were women. Clinical and laboratory data were obtained from 21 patients. Eighteen patients were febrile (15 required hospitalization); cellulitis was reported in 9 patients. Sixty percent were immunocompromised; 45% were reported to be seropositive for human immunodeficiency virus (HIV). For bacteremic patients, positive blood cultures were detected by a slightly elevated growth index in an automated blood culture system; many hospital laboratories had difficulty isolating the organism. CONCLUSIONS: Helicobacter cinaedi appears to cause recurrent cellulitis with fever and bacteremia in immunocompromised hosts. Blood cultures from immunocompromised patients with these symptoms may need special handling to isolate H. cinaedi.


Assuntos
Bacteriemia/epidemiologia , Celulite (Flegmão)/epidemiologia , Infecções por Helicobacter/epidemiologia , Helicobacter/isolamento & purificação , Hospedeiro Imunocomprometido , Adulto , Idoso , Idoso de 80 Anos ou mais , Bacteriemia/tratamento farmacológico , Bacteriemia/microbiologia , Celulite (Flegmão)/tratamento farmacológico , Celulite (Flegmão)/microbiologia , Feminino , Soropositividade para HIV , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Estados Unidos/epidemiologia
8.
Mol Cell Probes ; 8(2): 109-15, 1994 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-7935508

RESUMO

Restriction fragment length polymorphisms of ribosomal DNA (ribotyping) of many bacterial species has been useful for both epidemiologic subtyping and species identification. However, the use of ribotyping has been confined to major research and reference laboratories due to two factors: (a) the procedure must be carefully optimized for each organism one wishes to investigate and (b) most currently available protocols use hazardous chemicals or radioisotopes. The purpose of this study is to suggest an overall scheme that a clinical or research microbiologist could apply to ribotyping of any organism. In general, we recommend using a guanidium extraction method for DNA extraction, careful optimization of restriction conditions, and hybridization with non-radioactive digoxigenin-labelled probes; these procedures do not use hazardous chemicals or radioisotopes.


Assuntos
Campylobacter/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Helicobacter/genética , Southern Blotting , Campylobacter/classificação , DNA Bacteriano/análise , DNA Ribossômico/análise , Digoxigenina , Helicobacter/classificação , Técnicas Microbiológicas , Polimorfismo de Fragmento de Restrição
9.
J Infect Dis ; 168(3): 564-70, 1993 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8394859

RESUMO

In East Africa, bacteremia is more common in hospitalized human immunodeficiency virus (HIV) type 1-positive than -negative patients. In 1991, blood cultures and clinical and laboratory data were obtained from 319 patients in Ivory Coast, where both HIV-1 and -2 infections occur. Forty-three bacterial, 10 mycobacterial, and 8 fungal pathogens were isolated from blood of 54 patients (17%). Pathogens isolated significantly (P < or = .05) more frequently from HIV-positive than -negative patients were nonmycobacterial bacteria, particularly Salmonella enteritidis; mycobacteria, particularly Mycobacterium tuberculosis-Mycobacterium bovis; and yeast or fungus. HIV-1 or -2 positivity was associated with a 3-fold increased risk for septicemia (P < .02). HIV-positive patients with fever or with lymphocyte counts < 1000 were more likely to be septicemic than those without these characteristics. Mortality increased significantly with HIV positivity (40% vs. 14%, P < .001) and, among HIV-positive patients, with having pathogens isolated from blood (63% vs. 33%, P < .001).


Assuntos
Soropositividade para HIV/complicações , HIV-1 , HIV-2 , Sepse/complicações , Adulto , Bactérias/isolamento & purificação , Côte d'Ivoire/epidemiologia , Resistência Microbiana a Medicamentos , Feminino , Fungos/isolamento & purificação , Humanos , Masculino , Prognóstico , Sepse/microbiologia , Sepse/mortalidade
10.
Infect Immun ; 61(5): 2220-3, 1993 May.
Artigo em Inglês | MEDLINE | ID: mdl-8478115

RESUMO

After DNA hybridization identified an isolate from an ill rhesus macaque (Macaca mulatta) as Arcobacter (Campylobacter) butzleri, we initiated a study to determine whether A. butzleri was associated with diarrheal disease in nonhuman primates at the Yerkes Primate Research Center. By using Campy-CVA medium incubated at 35 degrees C, 15 A. butzleri isolates were obtained from 14 macaques; 7 macaques were coinfected with Campylobacter coli and Campylobacter jejuni. A. butzleri was not isolated from normal feces, despite the fact that feces from 76 macaques were cultured at necropsy. Histologic evaluation of colonic specimens from three macaques from which A. butzleri had been isolated showed mild to moderately severe chronic, active colitis. Ribotype analysis of the 15 A. butzleri isolates revealed nine different strains; these data suggest that A. butzleri may be endemic in this primate population and that a point source of infection is unlikely. This is the first report of the presence of A. butzleri in juvenile and adult macaques with diarrhea, and it may present an opportunity to study the pathogenesis of this organism, which appears to be associated with persistent diarrhea in humans.


Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter/patogenicidade , Diarreia/veterinária , Macaca/microbiologia , Animais , Campylobacter/genética , DNA Ribossômico/genética , Diarreia/microbiologia , Feminino , Masculino
11.
J Clin Microbiol ; 30(11): 3024-7, 1992 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1452677

RESUMO

Studies were conducted to characterize 18 isolates of Campylobacter spp. that could not be identified as either Campylobacter jejuni or C. coli. The isolates were cultured from specimens from 13 of 18 infant nonhuman primates during a prospective epidemiologic study reported previously. Phenotypic tests, DNA hybridization, and analysis of DNA coding for rRNA identified the isolates as C. butzleri (seven isolates), C. hyointestinalis (seven isolates), and C. fetus subsp. fetus or C. fetus subsp. fetus-like organisms (four isolates). Ribotype and polyacrylamide gel electrophoresis patterns indicated that there was heterogeneity among the isolates of C. butzleri and C. fetus subsp. fetus-like organisms.


Assuntos
Infecções por Campylobacter/veterinária , Doenças dos Macacos/microbiologia , Criação de Animais Domésticos , Animais , Proteínas de Bactérias/isolamento & purificação , Campylobacter/classificação , Campylobacter/genética , Campylobacter/isolamento & purificação , Infecções por Campylobacter/microbiologia , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , DNA Ribossômico/genética , DNA Ribossômico/isolamento & purificação , Macaca nemestrina
12.
Antimicrob Agents Chemother ; 36(4): 717-22, 1992 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-1503434

RESUMO

We evaluated the in vitro activities of 22 antimicrobial agents against 78 human and animal isolates belonging to two aerotolerant Campylobacter species, C. cryaerophila and C. butzleri, using a broth microdilution technique. An additional 10 antimicrobial agents were included at concentrations found in selective Campylobacter media. Strains of C. cryaerophila belonged to two DNA hybridization groups: DNA hybridization group 1A, which includes the type strain of C. cryaerophila, and DNA hybridization group 1B. The aminoglycosides, fluoroquinolones, and one tetracycline (minocycline) demonstrated the most activity against all DNA hybridization groups (C. cryaerophila DNA groups 1A and 1B and C. butzleri). Most isolates were resistant to cephalosporin antibiotics, with the exception of cefotaxime, and were variably susceptible to trimethoprim-sulfamethoxazole. C. cryaerophila DNA hybridization group 1A isolates were generally susceptible to the tetracyclines, chloramphenicol, nalidixic acid, azithromycin, erythromycin, and roxithromycin and moderately susceptible to clindamycin, trimethoprim-sulfamethoxazole, ampicillin, and ampicillin-sulbactam. The MICs of tetracyclines were higher for C. butzleri and C. cryaerophila DNA hybridization group 1B isolates than for C. cryaerophila DNA hybridization group 1A isolates, but most strains were still susceptible to doxycycline and tetracycline; all isolates were susceptible to minocycline. C. butzleri and C. cryaerophila DNA hybridization group 1B isolates were generally resistant to the macrolide antibiotics (including erythromycin), chloramphenicol, clindamycin, nalidixic acid, ampicillin, and trimethoprim-sulfamethoxazole. Differences in antimicrobial susceptibility between aerotolerant Campylobacter species and more common Campylobacter species, e.g., C. jejuni, suggest that different treatment strategies may be necessary. Strains of all three DNA hybridization groups of aerotolerant Campylobacter isolates were susceptible to colistin, polymyxin B, and rifampin at concentrations commonly used in selective media. These results suggest that primary isolation methods for Campylobacter species may need to be modified to include aerotolerant Campylobacter strains.


Assuntos
Antibacterianos/farmacologia , Campylobacter/efeitos dos fármacos , Animais , Campylobacter/genética , Humanos , Técnicas In Vitro , Testes de Sensibilidade Microbiana , Hibridização de Ácido Nucleico
13.
J Clin Microbiol ; 30(2): 362-9, 1992 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-1371517

RESUMO

Analysis of DNA restriction fragment length polymorphisms of rRNA genes (ribotyping) was employed to assist in the epidemiologic investigation of the emergence and spread of ciprofloxacin-resistant Staphylococcus aureus at the Atlanta VA Medical Center because many isolates of interest were nontypeable by phages and harbored few plasmids useful as strain markers. Chromosomal DNAs of selected S. aureus isolates were digested initially with 20 different restriction enzymes. EcoRI appeared to give the best discrimination of hybridization banding patterns (ribotypes) and was used with all study isolates. Overall, 15 different ribotypes were seen among the 50 S. aureus isolates studied (7 ribotypes among 13 methicillin-susceptible S. aureus [MSSA] isolates and 9 ribotypes among 37 methicillin-resistant S. aureus [MRSA] isolates). Seven of eight ciprofloxacin-resistant MSSA (CR-MSSA) patient isolates had identical antibiograms, were nontypeable by phages, and had a single 22-MDa plasmid. Six of these seven CR-MSSA isolates had an identical ribotype pattern. Ribotyping distinguished this CR-MSSA strain or clone from MRSA and other MSSA isolates, including nontypeable isolates that contained a 22-MDa plasmid. Five ciprofloxacin-susceptible MSSA isolates studied had five ribotypes; one pattern was identical to the CR-MSSA clone. Twenty-three CR-MRSA isolates recovered from the Atlanta VA Medical Center had four different ribotypes. Ribotyping proved to be a useful molecular epidemiologic tool in the study of S. aureus because it differentiated isolates which were indistinguishable by more traditional methods. In addition, this technique demonstrated that at our institution, ciprofloxacin resistance emerged in multiple strains of MRSA, as opposed to primarily a single strain or clone of MSSA.


Assuntos
Técnicas de Tipagem Bacteriana , Staphylococcus aureus/classificação , Bacteriófagos/classificação , Ciprofloxacina/farmacologia , DNA Bacteriano/genética , Resistência Microbiana a Medicamentos , Métodos Epidemiológicos , Estudos de Avaliação como Assunto , Genes Bacterianos , Humanos , Meticilina/farmacologia , Plasmídeos , Polimorfismo de Fragmento de Restrição , RNA Bacteriano/genética , RNA Ribossômico/genética , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética
14.
J Clin Microbiol ; 29(11): 2368-74, 1991 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1723068

RESUMO

Yersinia enterocolitica is a major enteric pathogen associated with a wide variety of clinical and immunologic manifestations, including transfusion-associated disease, from which there is a high mortality. Although previously rare in the United States, in the late 1980s Y. enterocolitica O:3 emerged as the predominant serotype in the United States, as it has been in Canada, Europe, and Japan. Epidemiologic investigation of this serogroup has been hampered by the limited availability of a phage typing system and the fact that Y. enterocolitica harbors few plasmids that are useful as strain markers. We therefore analyzed whole-cell DNA restriction fragment length polymorphisms of rRNA genes (ribotyping) to study a group of 61 (50 human, 11 porcine) Y. enterocolitica isolates. Initially, 20 different restriction enzymes were used: NciI appeared to give the best discrimination of hybridization banding patterns (ribotypes) within Y. enterocolitica O:3. Ribotyping distinguished seven clones among all the study isolates and four clones within Y. enterocolitica O:3 (53 isolates studied) and clearly differentiated Y. enterocolitica O:3 from Y. enterocolitica O:9; O:1,2,3; O:20; and O:5,27. Most serogroup O:3 isolates belonged to two clones, ribotypes I and II, including 23 of 24 Y. enterocolitica O:3 (13 human, 11 porcine chitterling) isolates recovered from a recent outbreak of Y. enterocolitica in children in Atlanta associated with chitterling preparation and 3 transfusion-associated O:3 isolates from the United States. Y. enterocolitica O:3 ribotypes I and II were also isolated in Japan, ribotypes II and IV were isolated in Belgium, and ribotype I was isolated in Canada. Ribotype patterns I and II corresponded to phage types 9b and 8, respectively. Ribotyping was able to distinguish individual strains of Y. enterocolitica O:3, but suggests that a limited number of clones have disseminated within the United States and globally. The finding of identical ribotype patterns in chitterling and human specimens from the Atlanta outbreak supports epidemiologic evidence that swine were the source of infection and major reservoir for Y. enterocolitica O:3.


Assuntos
DNA Bacteriano/genética , Yersiniose/epidemiologia , Yersinia enterocolitica , Animais , Técnicas de Tipagem Bacteriana , Surtos de Doenças , Reservatórios de Doenças , Métodos Epidemiológicos , Genes Bacterianos , Humanos , Polimorfismo de Fragmento de Restrição , RNA Bacteriano/genética , RNA Ribossômico/genética , Sorotipagem , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/microbiologia , Estados Unidos/epidemiologia , Yersiniose/microbiologia , Yersiniose/veterinária , Yersinia enterocolitica/classificação , Yersinia enterocolitica/genética , Yersinia enterocolitica/isolamento & purificação
15.
J Clin Microbiol ; 29(11): 2380-4, 1991 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1723069

RESUMO

Shigella sonnei is the most frequent cause of shigellosis in the United States. Epidemiologic studies of this organism have been hampered by the lack of adequate typing procedures. Ribosomal DNA analysis (ribotyping), a method which analyzes restriction fragment length polymorphisms in the chromosomal genes that encode rRNA, has recently been shown to be useful for microbial species identification and subtyping. To determine whether ribotyping could be used to distinguish between S. sonnei isolates, we conducted Southern hybridization studies on isolates from 16 different geographic locations and from four recent outbreaks. S. sonnei genomic DNA fragments generated following digestion with SalI hybridized with Escherichia coli 16S and 23S rRNAs to produce six distinct patterns; strains with patterns 1, 2, and 3 were each further subdivided into two additional patterns by using PvuII, SmaI, and SstI, respectively. Epidemiologically related strains had identical patterns. Ribotyping appears to be a useful tool for epidemiologic studies of shigellosis caused by S. sonnei.


Assuntos
RNA Bacteriano/genética , RNA Ribossômico/genética , Shigella sonnei/classificação , Shigella sonnei/genética , Técnicas de Tipagem Bacteriana , Surtos de Doenças , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/microbiologia , Humanos , Óperon , Polimorfismo de Fragmento de Restrição , Shigella sonnei/isolamento & purificação , Especificidade da Espécie , Estados Unidos
16.
J Clin Microbiol ; 29(8): 1670-6, 1991 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-1684797

RESUMO

Whole-cell chromosomal digests of 84 strains of aerotolerant Campylobacter (AC) were examined by using PvuII restriction fragment length polymorphisms of rRNA genes followed by hybridization with Escherichia coli 16S and 23S rRNA (ribotyping). The AC strains belonged to Campylobacter cryaerophila (n = 13) and a newly defined species, "C. butzleri" (n = 64). Strains of C. cryaerophila belonged to two hybridization groups: DNA group 1A (including the type strain of C. cryaerophila) and DNA group 1B (J. A. Kiehlbauch, D. J. Brenner, M. A. Nicholson, C. N. Baker, C. M. Patton, A. G. Steigerwalt, and I. K. Wachsmuth, J. Clin. Microbiol. 29:376-385, 1991). Six AC strains not classified as C. cryaerophila or "C. butzleri" were also included. All 35 sporadic human and animal isolates of "C. butzleri" sent to the Centers for Disease Control for identification showed different ribotype patterns. However, most "C. butzleri" strains contained common bands at approximately 3.0, 6.2, 12.0, and 15.0 kb; the 3.0-kb band was present in all but four strains. An additional 23 strains of "C. butzleri," isolated as part of special studies, contained the 3.0-kb band. Thus, on the basis of visual identification of the 3.0-kb band, 94% of available strains were correctly identified as "C. butzleri." Ribotyping demonstrated that C. cryaerophila strains (DNA groups 1A and 1B) were different from C. butzleri strains. All C. cryaerophila strains demonstrated a common ribosomal DNA restriction fragment of 3.2 kb; DNA group 1B strains contained an additional common band at 2.6 kb. Ribotyping patterns of AC species were easily distinguished from patterns of other Campylobacter, Helicobacter, and Wolinella species.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Campylobacter/genética , DNA Ribossômico/genética , Polimorfismo de Fragmento de Restrição , RNA Ribossômico/genética , Southern Blotting , Hibridização de Ácido Nucleico , Sondas RNA , RNA Ribossômico 16S/genética , RNA Ribossômico 5S/genética
17.
J Infect Dis ; 163(5): 1062-7, 1991 May.
Artigo em Inglês | MEDLINE | ID: mdl-2019754

RESUMO

Campylobacter species were isolated from 93 (15%) of 631 Thai children with diarrhea using the membrane filter technique on nonselective blood agar incubated at 37 degrees C. Campylobacter jejuni was isolated from 62 (10%), Campylobacter coli from 14 (2%), and atypical campylobacters from 17 (3%). The 17 atypical strains were first characterized biochemically and by dot blot DNA hybridization. Catalase-negative strains also were characterized by DNA hybridization and ribotype pattern. One strain was a catalase-negative "Campylobacter upsaliensis" and another was a nitrate-negative Campylobacter jejuni doylei. Fifteen isolates were aerotolerant strains most closely resembling Campylobacter cryaerophila or "C. upsaliensis" by dot hybridization. These aerotolerant strains, designated group 2 ("Campylobacter butzleri"), had ribotypes distinct from C. cryaerophila and have previously been shown to be related by DNA hybridization at the species level to the group 2 aerotolerant Campylobacter type strain (D2686). Group 2 aerotolerant Campylobacter were the atypical Campylobacter species most frequently isolated from Thai children with diarrhea.


Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter/isolamento & purificação , Diarreia/microbiologia , Campylobacter/classificação , Campylobacter/genética , Pré-Escolar , DNA Bacteriano/análise , Fezes/microbiologia , Humanos , Lactente , Hibridização de Ácido Nucleico , Fenótipo , Tailândia
18.
J Clin Microbiol ; 29(4): 680-8, 1991 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-1890168

RESUMO

We compared four phenotypic and six genotypic methods for distinguishing Campylobacter jejuni strains from animals and humans involved in four epidemics. Based on a comparison with epidemiologic data, the methods that correctly identified all strains in three milkborne outbreaks and one waterborne outbreak were heat-stable and heat-labile serotyping; multilocus enzyme electrophoresis (MEE); DNA restriction endonuclease analysis with BglII, XhoI, PvuII, or PstI; and Southern blot and hybridization of PvuII- and PstI-digested DNA with Escherichia coli 16S and 23S rRNA (ribotyping). Biotyping, phage typing, plasmid analysis, and probing of BglII and XhoI DNA digests with C. jejuni 16S rRNA genes failed to correctly separate one or more strains. MEE, restriction endonuclease analysis, and ribotyping were the most sensitive methods and identified nine types among the 22 strains. These methods were also capable of further distinguishing strains within the same serotype. Data from MEE were also analyzed to calculate genetic relatedness among strains. Serotyping was the most discriminating phenotypic method, with eight and seven types distinguished by the heat-stable and heat-labile methods, respectively. MEE and ribotyping had several advantages over the other methods because they measure relatively stable and significant chromosomal differences and are applicable to other species and genera. These methods, however, are complex and not easily quantified; they are currently limited to specialized laboratories. When antisera are available, serotyping appears to be an effective and more practical approach to the identification of epidemic-related strains.


Assuntos
Técnicas de Tipagem Bacteriana , Infecções por Campylobacter/epidemiologia , Campylobacter jejuni/classificação , Campylobacter/classificação , Animais , Aves , Southern Blotting , Campylobacter/genética , Campylobacter jejuni/genética , Surtos de Doenças , Eletroforese , Genótipo , Humanos , Lactente , Leite/microbiologia , Fenótipo , Filogenia , Plasmídeos , Reprodutibilidade dos Testes , Mapeamento por Restrição , Sensibilidade e Especificidade , Especificidade da Espécie , Estados Unidos/epidemiologia
19.
J Infect Dis ; 163(2): 406-9, 1991 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-1671055

RESUMO

During 1988 the number of Shigella dysenteriae type 1 infections reported in the United States increased fivefold. To determine if recent isolates from Mexico were related to those that caused epidemics of dysentery worldwide, Southern hybridization analysis was done with Shiga toxin and ribosomal RNA gene probes. Western hemisphere and Eastern Hemisphere strains differed by the size of a single EcoRI fragment carrying the Shiga toxin genes. Three ribosomal DNA (rDNA) patterns were observed, which correlated with the strain's continental origin for 81 of 83 isolates tested. Together the Shiga toxin and rDNA probe results indicated that recent Mexican isolates were chromosomally similar to earlier Central American isolates and distinct from Asian and African strains. This suggests there has been no significant exchange of organisms between continents in recent decades and that the 1988 outbreak in Mexico was caused by strains present in Central America since at least 1962.


Assuntos
Disenteria Bacilar/epidemiologia , Shigella dysenteriae/classificação , África/epidemiologia , Antibacterianos/farmacologia , Ásia/epidemiologia , Toxinas Bacterianas/genética , América Central/epidemiologia , DNA Ribossômico/análise , Surtos de Doenças , Disenteria Bacilar/microbiologia , Humanos , México/epidemiologia , Plasmídeos , Polimorfismo de Fragmento de Restrição , RNA Ribossômico/análise , Toxinas Shiga , Shigella dysenteriae/efeitos dos fármacos , Shigella dysenteriae/genética , Viagem , Estados Unidos/epidemiologia
20.
J Clin Microbiol ; 29(2): 376-85, 1991 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-2007646

RESUMO

Seventy-eight aerotolerant Campylobacter isolates were characterized phenotypically and by DNA hybridization (hydroxyapatite method at 50 and 65 degrees C). Two DNA relatedness groups were found. (i) Sixty-four strains belonged to aerotolerant Campylobacter DNA hybridization group 2. These organisms were isolated from humans, primarily with diarrheal illness, and animals on several continents. Strains were aerotolerant at 30 and 36 degrees C and catalase negative or weakly catalase positive, grew in media containing glycine and on MacConkey agar, were susceptible to nalidixic acid, and were resistant to cephalothin. The name Campylobacter butzleri sp. nov. is proposed for this group. (ii) DNA hybridization group 1 consisted of the type strain of Campylobacter cryaerophila and 13 additional strains isolated from 10 animals outside the United States and from three humans within the United States. This group was genetically diverse; five strains were closely related to the type strain of C. cryaerophila (DNA hybridization group 1A), and eight strains were more closely related to one another (DNA hybridization group 1B). Strains in DNA hybridization group 1B were phenotypically diverse, with two of eight strains resembling C. cryaerophila. The seven strains from DNA hybridization groups 1A and 1B which resembled C. cryaerophila and the C. cryaerophila type strain were aerotolerant only at 30 degrees C and catalase positive, did not grow in glycine or on MacConkey agar, were generally susceptible to nalidixic acid, and were resistant to cephalothin. The remaining six strains of DNA hybridization group 1B phenotypically resembled C. butzleri; however, they were generally catalase positive and susceptible to nalidixic acid and cephalothin. DNA hybridization group 1B is not designated as a separate species at this time since it cannot, with certainty, be separated genetically from C. cryaerophila or phenotypically from C. butzleri.


Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter/isolamento & purificação , Diarreia/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Campylobacter/classificação , Campylobacter/genética , Infecções por Campylobacter/veterinária , DNA Bacteriano/genética , Diarreia/veterinária , Genótipo , Humanos , Hibridização de Ácido Nucleico , Fenótipo , Especificidade da Espécie
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