Major Ribotype Research Articles

Ribotyping of Burkholderia pseudomallei from clinical and soil isolates in Thailand

Burkholderia pseudomallei is a soil saprophyte that causes melioidosis in humans and animals. Restriction fragment length polymorphism of the ribosomal DNA regions (ribotyping) were analyzed in 577 isolates comprising 371 clinical and 206 soil isolates collected throughout Thailand in 1997. A total of 77 distinct ribotype patterns consisting of 2–9 bands with sizes ranging from 2.8 to >23 kb were found. Twelve major ribotypes were identified of which types 3, 8 and 23 were commonly found (278/577, 48.2%) in both clinical (217/371, 58.5%) and environmental isolates (61/206, 29.6%). Three unique environmental types were found whereas a unique clinical type was not observed. Even though ribotypes show high heterogeneity in the rDNA region, the unique environmental patterns were clearly different from the clinical patterns as clearly seen by UPGMA dendrogram. Moreover, the three major types (types 3, 8 and 23) were discovered in nearly half of B. pseudomallei isolates. Subtyping of these major ribotypes in correlation with clinical profiles may help researchers to identify the virulence factor of the organism.

Characterization of paired mucoid/non-mucoid Pseudomonas aeruginosa isolates from Danish cystic fibrosis patients: antibiotic resistance, beta-lactamase activity and RiboPrinting.

The purpose of this study was to characterize 42 paired mucoid and non-mucoid Danish cystic fibrosis (CF) Pseudomonas aeruginosa isolates collected in 1997, by RiboPrinting, antibiotic susceptibility and beta-lactamase activity. Eight P. aeruginosa isolates collected before 1991 were included for comparison. Eighteen of the 42 paired mucoid and non-mucoid isolates showed the same ribotype; the remaining 24 belonged to different ribogroups. Mucoid isolates showed higher susceptibility to antibiotics and lower beta-lactamase activity compared with non-mucoid isolates. Significant differences (P < or = 0.01) between mucoid and non-mucoid isolates were found for the meropenem and colistin MICs for the isolates with the same ribotype, and for the MICs of ceftazidime, piperacillin, aztreonam, meropenem, tobramycin, ciprofloxacin and in the basal levels of beta-lactamase for the paired isolates belonging to different ribogroups. A dominant ribotype 73-S2 with hyperinducible beta-lactamase production and significantly higher MICs of piperacillin, meropenem and tobramycin compared with the other major ribotypes (73-S1, 207-S3 and 227-S8) was present among the 84 CF isolates. The isolates collected before 1991 had an antibiotic susceptibility pattern similar to the 1997 isolates. Despite prolonged and intensive antibiotic treatment, susceptible mucoid isolates were isolated from the CF sputum, possibly because these bacteria are protected from the selective pressure of antibiotics by the resistant non-mucoid isolates co-existing in the biofilm in the lungs of CF patients.

Ribotyping of Vibrio parahaemolyticus isolates obtained from food poisoning outbreaks in Taiwan.

Vibrio parahaemolyticus is a prevalent food-borne pathogen in Taiwan, Japan and other Asian countries. This work presents a novel ribotyping method for the molecular epidemiological examination of this pathogen. Genomic DNA was fragmented by HindIII digestion and hybridized with cDNA probe for Escherichia coli 16S and 23S RNA genes. A total of 121 isolates obtained from outbreaks during 1992 and 1994 in Taiwan were characterized by this ribotyping method. Four to seventeen restricted fragments were visualized in these isolates. After hierarchical cluster analysis, these isolates were grouped into thirty different ribotypes. In addition, A3, A7, E3 and F1 were the major ribotypes, consisting of 22.3, 13.2, 9.1, and 8.3% of the isolates, respectively. A, E, F, G and B were the major groups, consisting of 46.2, 14.0, 9.1, 6.7, and 6.7% of the isolates, respectively. The discriminatory ability of this ribotyping method, as determined by Simpson's index of diversity, was 0.93, which closely resembled that of a previously reported pulsed-field gel electrophoresis method.

Applicability of Ribotyping for Intraspecific Classification and Epidemiological Studies of Photobacterium damsela subsp. piscicida

Summary The ribotype analysis of a collection of twenty nine Photobacterium damsela subsp. piscicida strains, with different host and geographical origins, was performed in order to analyze the possible genetic variability within this bacterial fish pathogen. In addition, the value of this technique for intraspecific classification and epidemiological studies of Ph. damsela subsp. piscicida was evaluated. After restriction with four endonucleases, Pvu I Pvu II, Sma I, and Hind III, two major ribotypes could be established. The first ribopattern compiled the European strains, while the second ribotype included the Japanese isolates. A third ribotype was observed with the enzymes Pvu I, Sma I and Hind III, constituted in all cases by an unique strain. Endonuclease Pvu I clearly differentiated the french isolate, while Sma I and Hind III distinguished the Israeli strain from the other European Ph. damsela subsp. piscicida . The similarities among ribotypes, calculated by the Dice coefficient, varied from 30–40%, with Pvu I and Pvu II, to 80–90% with Sma I and Hind III. The results obtained indicate the usefulness of the ribotyping procedures for epidemiological studies and also as an important taxonomic tool for the fish pathogen Ph. damsela subsp. piscicida .