Superiority of Molecular Techniques for Identification of Gram-Negative, Oxidase-Positive Rods, Including Morphologically Nontypical
            <i>Pseudomonas aeruginosa</i>
            , from Patients with Cystic Fibrosis

Wellinghausen, Nele; Köthe, Juliane; Wirths, Beate; Sigge, Anja; Poppert, Sven

doi:10.1128/jcm.43.8.4070-4075.2005

Cited by 84 publications

(75 citation statements)

References 23 publications

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“…To date, the only published DNA probe [18] for the identification of Acinetobacter spp. is used under non-standard hybridization conditions and therefore cannot be used in concert with previously described diagnostic FISH-probes [10,11,13]. Another study describes…”

Section: Introductionmentioning

confidence: 99%

“…FISH works with fluorescence-labeled oligonucleotide probes, which bind to unique complementary target sites on ribosomal RNA [10][11][12][13]. The successful implementation of FISH has been described for detection and identification of various pathogens including Acinetobacter [10][11][12][13][14][15][16][17]. To date, the only published DNA probe [18] for the identification of Acinetobacter spp.…”

Section: Introductionmentioning

confidence: 99%

“…Fluorescence in situ hybridization (FISH) is an alternative molecular method for rapid identification of bacteria that is more cost-effective than automated NAAT platforms and requires less technical expertise to perform than most other available molecular methods. FISH works with fluorescence-labeled oligonucleotide probes, which bind to unique complementary target sites on ribosomal RNA [10][11][12][13]. The successful implementation of FISH has been described for detection and identification of various pathogens including Acinetobacter [10][11][12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

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Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Frickmann¹,

Essig²,

Hagen³

et al. 2011

European Journal of Microbiology and Immunology

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Multi-drug-resistant strains of the Acinetobacter baumannii complex cause nosocomial infections. Rapid identification of Acinetobacter spp. is desirable in order to facilitate therapeutic or hygiene decisions. We evaluated a newly designed DNA probe that can be used under standard conditions in both a microwave oven and a slide chamber for the rapid identification of Acinetobacter spp. by fluorescence in situ hybridization (FISH). Using FISH, the new probe correctly identified 81/81 Acinetobacter spp. isolates and excluded 109/109 tested non-target organisms from agar culture. Furthermore, the new probe correctly identified 7/7 Acinetobacter spp. in 214 blood cultures determined to contain Gram-negative bacteria by Gram staining. Using either the microwave oven or slide chamber technique, the new probe was able to identify Acinetobacter spp. in 100% of the samples tested. FISH used in conjunction with our newly designed probe provides an easy, cheap, precise, and rapid method for the preliminary identification of Acinetobacter spp., especially in laboratories where more sophisticated methods like matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) are not available.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Frickmann¹,

Essig²,

Hagen³

et al. 2011

European Journal of Microbiology and Immunology

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“…Identification of algicidal bacteria-To identify the algicidal bacterial isolates, bacterial chromosomal DNA was isolated according to the method described by Choi et al (2005), and molecular identification was performed as in Wellinghausen et al (2005). Briefly, 16S rDNA was amplified using primers 27F, 5′-GAGTTTGATCATGGCTCAG-3′ and 1492R, 5′-GGT-TACCTTGTTACGACTT-3′, in a 50 μL reaction volume containing 20 ng of template DNA, 1× PCR buffer (Sigma), 5 mM MgCl 2 , 10 mM dNTPs, 10 pM of each primer, and 2.5 units of Taq DNA polymerase.…”

Section: Materials and Proceduresmentioning

confidence: 99%

In situ bacterial mitigation of the toxic cyanobacterium Microcystis aeruginosa: implications for biological bloom control

Kim

Sang

Hwang

et al. 2008

Limnology & Ocean Methods

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The algicidal bacterium Xanthobacter autotrophicus HYS0201-SM02 (SM02) was isolated from the surface water of a eutrophic lake (Lake Daechung, Korea). In vivo and in situ experiments showed that SM02 had algicidal activity against both a cultured strain and natural colonial morphs of the toxic cyanobacterium, Microcystis aeruginosa. Both the SM02 bacteria and its culture filtrate showed anti-algal activity against M. aeruginosa, indicating that an algicidal substance was released from SM02. The threshold concentration of SM02 for maximal algicidal activity against a natural bloom of M. aeruginosa was 10 7 CFU/mL. In situ co-culture of SM02 and M. aeruginosa showed that SM02 did not benefit from the massive decay of M. aeruginosa. In fact, repeated inoculations with a low concentration of SM02 were required for optimal algicidal activity, suggesting that water quality worsened during co-culture (i.e., nutrients and microcystin-LR concentration increased). These results suggest a role for the algicidal bacterium X. autotrophicus SM02 in biorestoration but probably not in treating outdoor Microcystis blooms. When developing a biological agent to control M. aeruginosa blooms in the field, it will be important to screen for specific agents with low threshold concentrations and high algicidal activity.

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“…As Stenotrophomonas maltophilia species once belonged to the Pseudomonas genus, there are signifi cant diffi culties when trying to identify and clearly distinct between the Pseudomonas and Stenotrpohomonas genera. Results from the conventional systems, such as API 20 NE (bioMerieux, France) and BBL Crystal Enteric / Nonfermenter ID kit (Becton Dickinson, USA) are not considered authoritative in the scientifi c research fi eld because, even if they are categorized as "excellent" "very good" or "good identifi cation", these systems exhibit a high level of errors in differentiating between Pseudomonas and Stenotrophomonas species [4]. The most common cause of confusion with the classical identifi cation is discrepancy in the oxidase test in Stenotrophomonas species that are oxidase negative, but atypical, positive oxidase test occurs in as many as 20% of the strains [5].…”

Section: Introductionmentioning

confidence: 99%

Differentiation between Pseudomonas and Stenotrophomonas Species Isolated from Fish Using Molecular and MALDI-TOF Method

et al. 2016

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For the purpose of precise antibiotic susceptibility testing it is necessary to clearly distinguish Pseudomonas and Stenotrophomonas genera, considering acquired resistance of Pseudomonas species, as well as the intrinsic resistance of Stenotrophomonas species. This is why in the identifi cation of the 51 isolates originated from fi sh, the following methods were used: standard PCR, 16S rRNA gene sequencing, and MALDI-TOF.The results of the standard PCR test, 16S rRNA gene sequencing and MALDI-TOF analysis confi rmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confi rmed that 10 strains belong to Stenotrophomonas maltophilia species.Three strains were positive in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identifi ed these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS fi rst identifi ed these three strains as 99% Stenotrophomonas, and in the repeated identifi cation it identifi ed them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identifi ed these strains as Stenotrophomonas.Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identifi ed these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS fi rst identifi ed these three strains as 99% Stenotrophomonas, and in the repeated identifi cation it identifi ed them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identifi ed these strains as Stenotrophomonas.Although modern test methods that have very high specifi city (PCR, 16S rRNA gene sequencing, MALDI TOF) were used in this study, precise differentiation between Unauthenticated Download Date | 5/11/18 8:55 PM Aksentijević et al.: Differentiation between Pseudomonas and Stenotrophomonas species isolated from fi sh using molecular and MALDI-TOF method 305Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods.

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Superiority of Molecular Techniques for Identification of Gram-Negative, Oxidase-Positive Rods, Including Morphologically Nontypical Pseudomonas aeruginosa , from Patients with Cystic Fibrosis

Cited by 84 publications

References 23 publications

Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

Rapid identification ofAcinetobacterspp. by fluorescencein situhybridization (FISH) from colony and blood culture material

In situ bacterial mitigation of the toxic cyanobacterium Microcystis aeruginosa: implications for biological bloom control

Differentiation between Pseudomonas and Stenotrophomonas Species Isolated from Fish Using Molecular and MALDI-TOF Method

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