Comparison of PCR-based DNA fingerprinting techniques for the identification of Listeria species and their use for atypical Listeria isolates

Vaneechoutte, Mario; Tichy, Hans-Volker; Bannerman, Elizabeth; Jäger, B; Billé, Jacques

doi:10.1099/00207713-48-1-127

Cited by 97 publications

(59 citation statements)

References 43 publications

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“…Sequencing was carried out as described previously (Vaneechoutte et al, 2000) and similarity calculations and cluster analysis were carried out as described elsewhere (Nemec et al, 2001). Restriction digestion of amplified rDNA with HaeIII was carried out as described previously (Vaneechoutte et al, 1998). Phenotypic testing was carried out as described previously Laffineur et al, 2002).…”

Section: Methodsmentioning

confidence: 99%

Wautersia gen. nov., a novel genus accommodating the phylogenetic lineage including Ralstonia eutropha and related species, and proposal of Ralstonia [Pseudomonas] syzygii (Roberts et al. 1990) comb. nov.

Vaneechoutte

Kämpfer

Baère

et al. 2004

International Journal of Systematic and Evolutionary Microbiology

245

114

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Culture Collection of the University of Gøteborg, Gøteborg, SwedenComparative 16S rDNA sequence analysis indicates that two distinct sublineages, with a sequence dissimilarity of >4 % (bootstrap value, 100 %), exist within the genus Ralstonia: the Ralstonia eutropha lineage, which comprises Ralstonia basilensis, Ralstonia campinensis, R. eutropha, Ralstonia gilardii, Ralstonia metallidurans, Ralstonia oxalatica, Ralstonia paucula, Ralstonia respiraculi and Ralstonia taiwanensis; and the Ralstonia pickettii lineage, which comprises Ralstonia insidiosa, Ralstonia mannitolilytica, R. pickettii, Ralstonia solanacearum and Ralstonia syzygii comb. nov. (previously Pseudomonas syzygii ). This phylogenetic discrimination is supported by phenotypic differences. Members of the R. eutropha lineage have peritrichous flagella, do not produce acids from glucose and are susceptible to colistin, in contrast to members of the R. pickettii lineage, which have one or more polar flagella, produce acid from several carbohydrates and are colistin-resistant. Members of the R. pickettii lineage are viable for up to 6 days on tryptic soy agar at 25 6C, whereas members of the R. eutropha lineage are viable for longer than 9 days. It is proposed that species of the R. eutropha lineage should be classified in a novel genus, Wautersia gen. nov. Finally, based on the literature and new DNA-DNA hybridization data, it is proposed that Pseudomonas syzygii should be renamed Ralstonia syzygii comb. nov. INTRODUCTIONThe genus Ralstonia (Yabuuchi et al., 1995) was created to accommodate bacteria from ecologically diverse niches that were classified previously as Burkholderia (Yabuuchi et al., 1992) and Alcaligenes. The type species of the genus -Ralstonia pickettii (type strain, ATCC 27511 T ) -was regarded originally as the only representative of clinical importance (Fass & Barnishan, 1976;Fujita et al., 1981;Kahan et al., 1983;Gardner & Shulman, 1984;Verschraegen et al., 1985;Roberts et al., 1990a;Lacey & Want, 1991; Dimech et al., 1993;Raveh et al., 1993) In a previous study , it was indicated that species of the genus Ralstonia can be separated clearly into two phenotypically and genotypically distinct groups. Here, it is proposed to consolidate these findings by allocating one group of species to a novel genus, Wautersia The results of lactose and maltose acidification by Ralstonia pickettii strains, the 16S rRNA gene signature sequences of the Ralstonia species, detailed phenotypic data of different species and a similarity matrix of the 16S rDNA sequences are available as supplementary material (Tables A1, A2, A3 and A4, respectively) in IJSEM Online.

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Section: Methodsmentioning

confidence: 99%

Wautersia gen. nov., a novel genus accommodating the phylogenetic lineage including Ralstonia eutropha and related species, and proposal of Ralstonia [Pseudomonas] syzygii (Roberts et al. 1990) comb. nov.

Vaneechoutte

Kämpfer

Baère

et al. 2004

International Journal of Systematic and Evolutionary Microbiology

245

114

View full text Add to dashboard Cite

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“…Isolates were identi®ed to species level using tDNAintergenic polymerase chain reaction (tDNA-PCR) (Welsh and McClelland 1991) in combination with capillary electrophoresis (Vaneechoutte et al 1998;Baele et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Composition of enterococcal and streptococcal flora from pigeon intestines

et al. 2002

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Aims: To identify the streptococcal and enterococcal components of the micro¯ora from pigeon intestines to species level by tDNA-intergenic polymerase chain reaction. Methods and Results: Gram-positive coccal strains were isolated from the intestines or faeces of 50 different pigeons, all from different lofts, on Columbia CNA blood agar. Sodium azidesensitive enterococci were found to constitute the major coccal members of the intestinal¯ora of pigeons. Forty-two of 69 isolates were identi®ed as Enterococcus columbae (84% of animals tested positive), while E. cecorum was encountered in 28% of all animals (14 isolates). Minor species (less than 10% of pigeons positive) were E. faecalis, E. faecium, E. gallinarum, E. casseli¯avus, Streptococcus alactolyticus and Strep. gallolyticus. Contrary to the species description of E. columbae and E. cecorum, some of the strains were found to be motile. Certain other minor differences with the species description of E. columbae are also indicated. Conclusions: Enterococcus columbae is the major Gram-positive facultatively anaerobic component of the pigeon intestinal¯ora. Certain phenotypic characteristics differ from the original species description. The potentially pathogenic Strep. gallolyticus is a minor component of the micro¯ora. Enterococcus faecium and E. faecalis are rare in pigeons. Signi®cance and Impact of the Study: The enterococcal and streptococcal¯ora of pigeons is composed of mainly host-speci®c bacteria. These are not likely to act as zoonotic pathogens nor as carriers of antibiotic resistance determinants possibly spreading to humans.

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“…Using primers aimed at the amplification of tRNA intergenic spacer regions (1,4,5), no amplification signal could be obtained, as is the case for most Ralstonia species (unpublished results). Therefore, tRNA-PCR appears not to be useful for the identification of most Ralstonia species.…”

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confidence: 99%

Case of Catheter Sepsis with Ralstonia gilardii in a Child with Acute Lymphoblastic Leukemia

Wauters

Claeys²,

Verschraegen³

et al. 2001

J Clin Microbiol

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Acute lymphoblastic leukemia was diagnosed in a 7-year-old girl. Two months after insertion of a central venous catheter, she developed fever and complained of headache and abdominal pain. Physical examination revealed no focus of infection. A gram-negative nonfermenting bacillus was recurrently cultured from blood. Extensive biochemical testing and 16S ribosomal DNA sequencing led to the identification of Ralstonia gilardii. CASE REPORTAcute lymphoblastic leukemia (ALL) was diagnosed in a 7-year-old girl in May 2000, and treatment was initiated according to the EORTC-CLCG-58951 protocol for children with very-low-risk ALL. The girl achieved hematologic remission after induction chemotherapy. A central venous catheter was inserted in June 2000. The girl tolerated the treatment uneventfully until September 2000, when, during a course of chemotherapy (high-dose methotrexate), she developed spiking fever as high as 40°C. She complained of headache and abdominal pain and vomited twice. Physical examination revealed no focus of infection. The leucocyte count was 5,800/ml, with an absolute neutrophil count of 4,760/ml and elevated C-reactive protein (87 mg/dl). The chemotherapy was stopped, and the girl was treated with intravenous (i.v.) ampicillin (100 mg/kg of body weight/day). One day later blood cultures grew gram-negative bacilli, and (i.v.) netromycin (7.5 mg/kg/day) treatment was added. The spiking fever disappeared and the girl's health improved.A gram-negative nonfermenting bacillus was isolated and found to be resistant to ampicillin, piperacillin, aztreonam, gentamicin, and tobramycin and susceptible to cefuroxime, ceftriaxone, ceftazidime, imipenem, co-trimoxazole, ofloxacin, and amikacin. The girl was treated as an outpatient with i.v. ceftriaxone (100 mg/kg/day) once daily for 4 more days. Thirtysix hours after the ceftriaxone treatment was stopped, she again developed spiking fever, and i.v. ceftriaxone (100 mg/kg/ day) was restarted in combination with i.v. amikacin (15 mg/ kg/day). Again, a gram-negative nonfermenting bacillus was cultured from blood. The girl showed an allergic reaction to ceftriaxone with rash and pruritus, and the ceftriaxone was replaced with i.v. ciprofloxacin (20 mg/kg/day). The spiking fever disappeared again, and amikacin and ciprofloxacin i.v. treatment was given for 7 more days. Blood cultures remained negative, and the central venous catheter was not removed. Three months later the intensive chemotherapy was completed and the girl was doing well. The catheter was removed on January 8 2001. No nonfermenting gram-negative bacilli were cultured from the tip.Discussion. The gram-negative bacillus was isolated in pure culture from all eight FAN aerobic blood cultures and from one of the eight BacT/Alert anaerobic cultures (Organon Teknika, Turnhout, Belgium), collected over a period of 10 days. Initial identification based on the API20NE system (bioMérieux, Marcy l'Etoile, France) yielded code 1000474, leading to an identification as Alcaligenes faecalis, Comamonas acidovorans ...

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Comparison of PCR-based DNA fingerprinting techniques for the identification of Listeria species and their use for atypical Listeria isolates

Cited by 97 publications

References 43 publications

Wautersia gen. nov., a novel genus accommodating the phylogenetic lineage including Ralstonia eutropha and related species, and proposal of Ralstonia [Pseudomonas] syzygii (Roberts et al. 1990) comb. nov.

Wautersia gen. nov., a novel genus accommodating the phylogenetic lineage including Ralstonia eutropha and related species, and proposal of Ralstonia [Pseudomonas] syzygii (Roberts et al. 1990) comb. nov.

Composition of enterococcal and streptococcal flora from pigeon intestines

Case of Catheter Sepsis with Ralstonia gilardii in a Child with Acute Lymphoblastic Leukemia

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