Evaluation of three procedures using a laser densitometer and microcomputer for estimating molecular sizes of restriction endonuclease digest fragments and application to Campylobacter jejuni chromosomal DNA

Owen, Robert J.; Beck, A.

doi:10.1111/j.1472-765x.1987.tb01569.x

Cited by 23 publications

(9 citation statements)

References 8 publications

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“…Band size estimation Fragment sizes in the total digest and in the Southern blot hybridization patterns were calculated from migration distances by the DNA SIZE program as described previously [26]. (Fig.…”

Section: Blotting and Hybridizationmentioning

confidence: 99%

DNA restriction digest and ribosomal RNA gene patterns ofCampylobacter jejuni: a comparison with bio-, sero-, and bacteriophage-types of United Kingdom outbreak strains

1990

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SUMMARYDNA restriction endonuclease (Hae III and Hind III) total digest and 16S and 23S ribosomal (r)RNA gene patterns (ribopatterns) were determined for 18 isolates of Campylobacter jejuni from three separate outbreaks of diarrhoea in the north of England. Strains were also characterized by biotyping, serotyping and phage typing. Comparisons of the DNA patterns by visual and numerical methods revealed five distinct strain groupings with clear differences between isolates from different outbreaks as well as some heterogeneity between strains within the community outbreak and one of the school outbreaks. An excellent correlation was observed between the genomic DNA fingerprints data and the Preston bacteriophage group, both of which gave better discrimination than biotyping and serotyping alone or in combination. Only one phage group (PG 37) was not confirmed by the DNA data. DNA fingerprints therefore provide additional information of value in studying the epidemiology of outbreaks of C. jejuni.

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Section: Blotting and Hybridizationmentioning

confidence: 99%

DNA restriction digest and ribosomal RNA gene patterns ofCampylobacter jejuni: a comparison with bio-, sero-, and bacteriophage-types of United Kingdom outbreak strains

1990

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“…Phage typing has been developed but utilized in only a limited number of studies on C. jejuni [13][14][15]. Similarly, DNA (molecular) typing, based on total genomic DNA digest patterns from conventional [14,[16][17][18][19] and pulse field gel electrophoresis [20], ribosomal RNA gene restriction digest patterns [14,17,[21][22][23][24], and plasmid profiling [25], has been applied to a limited extent. Recently, the polymerase chain reaction (PCR) was used to obtain randomly amplified polymorphic DNA (RAPD) profiles.…”

Section: Introductionmentioning

confidence: 99%

Molecular subtyping by genome and plasmid analysis ofCampylobacter jejuniserogroups O1 and O2 (Penner) from sporadic and outbreak cases of human diarrhoea

Fayos

Owen²,

Hernández³

et al. 1993

Epidemiol. Infect.

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SUMMARYRibosomal RNA gene patterns, randomly amplified polymorphic genomic DNA (RAPD) profiles and plasmid profiles were used to discriminate between 28 strains of Campylobacterjejuni serogroups 01 and 02 (Penner). Most isolates were biotype I (Lior). The strains were representative isolates from a UK school outbreak of enteritis (7 cases) and from 21 sporadic human cases of enteritis in 4 countries. The molecular techniques discriminated to various degrees between strains in each of the serogroups. The outbreak strains were homogeneous in most molecular features but a variety of types was detected amongst the isolates from the sporadic cases. Five groups of two or more strains with identical ribopatterns were identified and within each, strains from different patients were homogenous with respect to serogroup. RAPD profile typing based on numerical analysis generally matched ribotyping. Plasmid profiling overall gave least discrimination but was useful in separating some strains similar in other features. We concluded that optimal discrimination of C. jejuni could best be achieved using a combination of phenotypic and genotypic properties. Hae III ribotyping was the single most discriminatory and reproducible technique investigated. Several strains of C. jejuni from sporadic infections had similar molecular profiles which have potential for general typing purposes.

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“…Double bands falling within a given range were scored as a single band. Computed similarities among strains were estimated by means of the Dice coefficient (negative matches excluded) and clustering of strains was based on the unweighted pair group method [19]. H. pylori NCTC 11637 was included as a reference on each gel.…”

Section: Computation Of Strain Similaritiesmentioning

confidence: 99%

Ribosomal RNA gene patterns ofHelicobacter pylorifrom surgical patients with healed and recurrent peptic ulcers

Owen¹,

Bickley²,

Lastovica

et al. 1992

Epidemiol. Infect.

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SUMMARYFifty-two strains of Helicobacter pylori were examined by DNA restriction endonuclease digestion, ribosomal (r)RNA gene probe hybridization and biotyping. Most (49) strains originated from gastric (antral) biopsies of patients before or after elective surgery for duodenal ulcers. Chromosomal DNA Hind III ribopatterns showed 9 strain clusters of which the largest contained 12 strains each with 3 common bands (1 50, 3 45, and 4 26 kb) but which were heterogeneous with respect to biotype and total digest pattern. Isolates from post-operative patients with either healed or recurrent ulcers showed ribopattern heterogeneity and exhibited a similar distribution of H. pylori ribopattern types; no single type predominated in any patient group or was more highly associated with recurrent ulcers than with healed ulcers. Multiple isolates from two surgical patients had only minor genomic variations in each set whereas isolates from two brothers had different ribopatterns. We conclude that Hind III ribopatterns in conjunction with total digest patterns might provide the basis for future epidemiological typing studies.

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Evaluation of three procedures using a laser densitometer and microcomputer for estimating molecular sizes of restriction endonuclease digest fragments and application to Campylobacter jejuni chromosomal DNA

Cited by 23 publications

References 8 publications

DNA restriction digest and ribosomal RNA gene patterns ofCampylobacter jejuni: a comparison with bio-, sero-, and bacteriophage-types of United Kingdom outbreak strains

DNA restriction digest and ribosomal RNA gene patterns ofCampylobacter jejuni: a comparison with bio-, sero-, and bacteriophage-types of United Kingdom outbreak strains

Molecular subtyping by genome and plasmid analysis ofCampylobacter jejuniserogroups O1 and O2 (Penner) from sporadic and outbreak cases of human diarrhoea

Ribosomal RNA gene patterns ofHelicobacter pylorifrom surgical patients with healed and recurrent peptic ulcers

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