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PubMed1991

Plasmid analysis of Mycobacterium avium-intracellulare (MAI) isolated in the United Kingdom from patients with and without AIDS.

One hundred and forty-seven isolates (128 strains) of Mycobacterium avium-intracellulare (MAI) were screened by agarose gel electrophoresis for the presence of plasmids. Plasmids were characterised according to size and by Southern hybridisation analysis of intact and restriction endonuclease-digested DNA. Two cloned MAI plasmids, pLR7 and pLR20, were used as probes. There was no significant difference in the rate of plasmid carriage in MAI strains isolated from patients with the acquired immuno-deficiency syndrome (AIDS) and from non-AIDS patients in the UK, but a higher rate of plasmid carriage was observed in a panel of American strains from AIDS patients. Plasmids were grouped into two broad categories: small (mostly 14-30 kb) and large (greater than 150 kb). Southern blot analysis identified two distinct groups of small plasmids, the majority of which showed homology with pLR7. Plasmids from this group were significantly more common in strains of serotypes 4 and 8 which are particularly associated with AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1994

Genetic and molecular R-plasmid analysis of Enterobacteriaceae hospital strains at Children's Hospitals of the former USSR.

R-plasmids from Enterobacteriaceae clinical strains, mainly Klebsiella and Serratia, isolated at different neonatal and children's hospitals of different cities of the former USSR for 10 years, were studied for their possible influence on the bacterial host phenotype. Hospital R-plasmids of stable inheritance persisted in hospitals from 2 to 7 years and were disseminated among strains of different genera (Klebsiella, Serratia, Enterobacter) and among different units. The data showed a possibility of long-term molecular rearrangements of R-plasmids in the hospital settings and an acquisition of genetic determinants encoding enterotoxin production. A novel R-plasmid encoding cytotoxicity to HEp-2 cells involved in two nosocomial outbreaks due to K. pneumoniae strains was reported. K. pneumoniae population heterogeneity was evaluated by using the plasmid parameters of strains. Their heterogeneity of a bacterial population was significantly lower during nosocomial outbreaks than in interepidemic periods.

Base Sequence↗
PubMed1987

Rapid plasmid analysis for identification of Edwardsiella ictaluri from infected channel catfish (Ictalurus punctatus).

Eighteen different strains of Edwardsiella ictaluri isolated from infected channel catfish (Ictalurus punctatus) were screened to determine whether plasmid DNA was present. Two plasmids of 5,700 and 4,900 base pairs were identified. Restriction enzyme analysis showed that each of the strains harbored these same two plasmids. Restriction maps of the separated plasmids indicated that these plasmids were not closely related to each other. A rapid screening technique was developed that would allow the presence of these plasmids from either broth cultures or single colonies of E. ictaluri to be determined within 2 to 3 h by agarose gel electrophoresis. These results suggest that plasmid fingerprinting of E. ictaluri should become a useful tool in the presumptive identification of this bacterium from infected channel catfish.

Animals↗
PubMed1991

[Plasmid analysis of antibiotic resistant Shigella isolates].

A correlation between the multiple drug resistance patterns and the plasmid profiles given by 70 clinical isolates of Shigella sonnei and Shigella flexneri was investigated in this study. The most common plasmids were purified from different isolates by electroelution and characterized via restriction endonuclease digestions.

Anti-Bacterial Agents↗
PubMed1998

Identification and characterization of Arcobacter isolates from broilers by biochemical tests, antimicrobial resistance patterns and plasmid analysis.

A total of 89 Arcobacter butzleri isolates obtained from freshly slaughtered broilers was investigated for their biochemical capacities by using the API CAMPY system as well as additional growth and tolerance tests, for their antimicrobial resistance patterns and their plasmid content. Among these techniques, biochemical profiling and antimicrobial resistance testing proved to be the most discriminatory methods as confirmed by the calculation of discriminatory indices. These two methods might represent valuable tools for epidemiological analysis of A. butzleri isolates. Small plasmids of up to 5 kbp were detectable in only 21 of the 89 isolates while the remaining 68 A. butzleri isolates were plasmid-free.

Abattoirs↗
PubMed1980

Characterization of promoter-cloning plasmids: analysis of operon structure in the rif region of Escherichia coli and isolation of an enhanced internal promoter mutant.

Using the promotor-cloning vehicle described by An and Friesen (J. Bacteriol. 140:400-410, 1979), Escherichia coli chromosomal deoxyribonucleic acid fragments derived from the lambda drifd18 transducing phage were cloned in one of several unique restriction endonuclease sites adjacent to tetracycline(tet) genes that lack their own promotor. One of these plasmids has been used to isolate nine variants having mutations that lie in a putative internal promoter which is located between rplL and rpoB. Deoxyribonucleic acid sequence analysis revealed that, in all nine mutants, a single base change, C to T, in the ribonucleic acid polymerase recognition site led to a large increase in promoter activity. Analysis of a variety of plasmids in which tet is fused to various promoters yielded the following results: (i) rplK and rplA, genes for ribosomal protein L11 and L1, respectively, were cotranscribed from a common promoter located upstream from rplK; (ii) there was a strong promoter in the region between the rplKA operon and rplJ, the gene for ribosomal protein L10; (iii) an attenuator region was located between rplL, the gene for ribosomal protein L12, and rpoB, the gene for ribonucleic acid polymerase subunit beta; (iv) transcription terminated immediately after rpoC, the gene for ribonucleic acid polymerase subunit beta'; (v) a gene coding for unknown protein U, which is located between tufB and the rplKA operon, had its own promoter; (vi) the tufB gene was separated from all of the genes described above and had its own promoter.

Bacterial Proteins↗
PubMed1985

In vitro antimicrobial susceptibility, plasmid analysis, and serotyping of epidemic-associated Campylobacter jejuni.

Campylobacter jejuni strains from 11 outbreaks were characterized by antimicrobial susceptibility, plasmid profile, and serotyping by the methods of Lior et al. and Penner and Hennessy. All 31 strains were susceptible to erythromycin, clindamycin, chloramphenicol, kanamycin, tobramycin, streptomycin, and gentamicin. A total of 21 strains from nine outbreaks were resistant to one or more of the following antimicrobial agents: tetracycline, metronidazole, ampicillin, or carbenicillin. Of the 31 strains, 19 possessed plasmid DNA; 4 of the strains containing plasmids were sensitive to all antimicrobial agents tested. All of the strains that were resistant to tetracycline contained a 38-megadalton plasmid, and these plasmids shared common nucleic acid sequences. No other antimicrobial resistance was associated with the presence of plasmid DNA. Eight outbreaks appeared to have been caused by a single serotype, whereas in three outbreaks multiple serotypes were found. In two of the three outbreaks with multiple serotypes, plasmid profiles were also indicative of multiple strains of C. jejuni. Antimicrobial susceptibility and plasmid profile are potentially useful epidemiological markers for C. jejuni and may be used to supplement serotyping.

Animals↗
PubMed1992

Success of DNA fingerprinting after failure of biotyping, antimicrobial susceptibility testing, and plasmid analysis to reveal clonality of multiple blood and urine isolates from a patient with Escherichia coli urosepsis.

Multiple isolates of Escherichia coli from the blood and urine of a 60-year-old woman with acute pyelonephritis exhibited different biotypes, antimicrobial susceptibility patterns, and plasmid profiles, suggesting the presence of polymicrobial bacteriuria and leaving in question the origin of the bacteremia. Only after bacterial restriction endonuclease analysis of total bacterial DNA was it discovered that all isolates represented the same strain, with plasmid instability possibly accounting for the varied antimicrobial susceptibility patterns observed. We conclude that the biotype, antimicrobial susceptibility profile, and plasmid profile are sometimes inadequate to clarify the relationships between different clinical isolates of E. coli from a single patient and can lead to erroneous epidemiologic conclusions. DNA fingerprinting can resolve dilemmas these less precise techniques leave unresolved.

Anti-Bacterial Agents↗
PubMed1988

Plasmid analysis and cloning of the dichloromethane-utilization genes of Methylobacterium sp. DM4.

The dichloromethane (DCM)-utilizing facultative methylotroph Methylobacterium sp. DM4 was shown to contain three plasmids with approximate size of 120 kb, 40 kb and 8 kb. Curing experiments suggested that the DCM-utilization character was correlated with the possession of an intact 120 kb plasmid. The DCM-utilization genes were cloned on the broad-host-range vector pVK100. Plasmid pME1510, a recombinant plasmid carrying a 21 kb HindIII fragment complemented DCM-utilization-negative derivatives of Methylobacterium sp. DM4 and conferred the DCM-utilization-positive phenotype to a number of Gram-negative methylotrophic bacteria. In Southern hybridization experiments with pMe1510 as a probe, chromosomal DNA from Methylobacterium sp. DM4 gave definite signals while purified plasmid DNA did not. Plasmid pME1510 did not hybridize with total DNA from a cured DCM-non-utilizing derivative of Methylobacterium sp. DM4. It is concluded that the DCM-utilization genes are located on the chromosome or on a megaplasmid. Curing procedures thus led to the formation of a chromosomal or megaplasmid deletion larger than 21 kb and covering the DCM-utilization genes or to the loss of an undetected megaplasmid.

Chromosomes, Bacterial↗
PubMed1989

Plasmid analysis of 219 methicillin-resistant Staphylococcus aureus strains with uncommon profiles isolated in Lisbon.

During the years 1986 and 1987, 219 methicillin-resistant Staphylococcus aureus (MRSA) strains were isolated in a Lisbon hospital. Antimicrobial susceptibilities and genetic analysis showed that resistance to penicillin, methicillin, erythromycin (inducible phenotype), tetracycline, gentamicin, tobramycin, kanamycin, streptomycin, neomycin and trimethoprim were chromosomally encoded. Plasmid DNA was absent from 38.8% of the isolates. Constitutive erythromycin resistance was coded by three plasmids of c. 2.3 Md, c. 2.0 Md and c. 1.6 Md. Chloramphenicol resistance was mediated by two plasmids of c. 2.9 Md and c. 1.8 Md. Small cryptic plasmids of c. 1.65 Md, c. 1.2 Md and c. 1.0 Md were also detected. The majority of the strains revealed antigen 17, were lysed by phages 75, 89 and/or 85, and were either devoid of plasmid DNA, or possessed a c. 1.6 Md plasmid coding for constitutive erythromycin resistance or a c. 1.0 Md cryptic plasmid. These observations cannot rule out that the MRSA Lisbon isolates are a homogeneous group of strains that might have a common origin, and seem to be different from MRSA recently isolated in other countries.

DNA, Bacterial↗
PubMed1986

Development of antibiotic resistance by staphylococcus aureus in a single patient. Confirmation by phage typing, antibiograms, and plasmid analysis.

A patient is described who had colonization and infection with strains of Staphylococcus aureus increasingly resistant to multiple antibiotics while receiving several courses of broad-spectrum antibiotics. S. aureus strains with different phage types, antibiograms, and plasmid profiles were isolated from different sites at the same time. After the patient was transferred to an intermediate-care service where he received only supportive care and no further antibiotics, culture samples from areas previously colonized or infected (sputum, anterior nares, perineal region, rectum) showed no S. aureus. This patient graphically demonstrates that antibiotics may create an environment allowing the overgrowth of resistant organisms and that withdrawal of this selection pressure may allow more sensitive organisms to recolonize.

Adult↗
PubMedDate not supplied

Plasmid analysis and epidemiology of Salmonella enteritidis infection in three commercial layer flocks.

Ninety-six S. enteritidis isolates obtained from three commercial layer flocks in 1988-90 were examined following DNA extraction, restriction enzyme digestion, and gel electrophoresis for plasmid size profiles and restriction fragment length polymorphisms (RFLPs). The S. enteritidis isolates from the three flocks had three, eight, and two different plasmid profiles, respectively. Only four isolates from one flock lacked plasmids. A 36-megadalton (mDa) (54-kilobase) plasmid was present in 73% of the isolates, either alone or in combination with other plasmids. Isolates with only the 36-mDa plasmid had identical RFLPs. The diversity of plasmid profiles was greater than that of phage-types among isolates from the three flocks: 12 unique plasmid profiles vs. four phage-types. Mixed infections with S. enteritidis strains having distinct plasmid profiles occurred in all three flocks. Reinfection of these flocks in 1990 with one or more of the strains obtained earlier was evident, because some of the original isolates and the 1990 isolates had matching plasmid profiles and were of the same phage-types. Isolates from both environmental and tissue samples, examined from one flock, were found to share the same plasmid profile and phage-type.

Animals↗
PubMed1992

Studies on an outbreak of neonatal diarrhea caused by EPEC 0127:H6 with plasmid analysis restriction analysis and outer membrane protein determination.

An outbreak of enteropathogenic Escherichia coli (EPEC) 0127:H6 diarrhea occurred at two nurseries for the newborn in Chongqing in May 1987. Sixty-nine neonates had diarrhea; two deaths resulted. The epidemic strains, carrying 1.5 and 60 Md plasmid DNA, had an identical restriction digest profile and the same outer membrane protein pattern and could produce localized adherence to HeLa, HEp-2 and FL cells. The rates of contamination with EPEC 0127:H6 on medical staff's hands in these two nurseries were 11.8% and 8.7%, respectively, whereas 85 samples from milk, air and other sources were all negative for EPEC. The source of infection was the index case's mother who had had watery stools. Transmission of EPEC 0127:H6 from infant to infant took place by way of the fecal-oral route, most likely via the hands of medical staff attending their care. We present the first case, confirmed by plasmid and restriction analyses and outer membrane protein determination, of a neonate who acquired EPEC during delivery through ingestion of organisms residing in the maternal birth canal.

Bacterial Outer Membrane Proteins↗