Differentiation between Pseudomonas and Stenotrophomonas species isolated from fish using molecular and MALDI-TOF method
Diferencijacija Pseudomonas i Stenotrophomonas vrsta izolovanih iz riba primenom molekularnih metoda i MALDI-TOF metode
2016
Authors
Aksentijević, KsenijaAšanin, Jelena
Milivojević, Dušan
Čolović, Svetlana
Butorac, Ana
Cindrić, Mario
Mišić, Dušan
Article (Published version)
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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 identification of the 51 isolates originated from fish, 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 confirmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confirmed 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 identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified ...them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Although modern test methods that have very high specificity (PCR, 16S rRNA gene Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods.
S obzirom na značaj stečene rezistencije Pseudomonas vrsta, kao i na intrinzičnu rezistenciju Stenotrophomonas vrsta, a u cilju preciznog ispitivanja osetljivosti na antibiotike, neophodna je jasna diferencijacija pripadnika ovih rodova bakterija. U tom cilju su u identifikaciji 51 izolata poreklom od riba korišćene metode: standardni PCR, 16S rRNA sekvenciranje gena, MALDI-TOF. Rezultati standardnog PCR testa, 16S rRNA sekvenciranja gena i MALDI-TOF analize su za 35 sojeva potvrdili pripadnost rodu Pseudomonas. Standardnim PCR testom i primenom aparata VITEK MS utvrđeno je da 10 sojeva pripada vrsti Stenotrophomonas maltophilia. U 16S rRNA sekvenciranju gena 3 soja koja su bila pozitivna u oba standardna PCR testa identifikovana su kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. VITEK MS je ova tri soja u prvoj identifikaciji identifikovao kao 99% Stenotrophomonas, a u ponovljenoj identifikaciji kao 99% Pseudomonas. Ti sojevi su na aparatu MALDI TOF/TOF 4800 Plus bili identifikovan...i kao Stenotrophomonas. U 16S rRNA sekvenciranju gena tri soja koja su bila negativna u oba standardna PCR testa su identifikovana kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. Aparat VITEK MS je ova tri soja identifikovao u jednoj identifikaciji kao 99% Stenotrophomonas, a u drugoj identifikaciji 99% kao Pseudomonas. Ti sojevi su na aparatu MALDI TOF/ TOF 4800 Plus bili identifikovani kao Stenotrophomonas. Iako su u ovom istraživanju korišćene savremene metode ispitivanja koje imaju vrlo visoku specifičnost (PCR, 16s rRNK sequencing, MALDI TOF) precizna diferencijacija Pseudomonas i Stenotrophomonas vrsta nije mogla biti postignuta.
Keywords:
fish / Pseudomonas / Stenotrophomonas / PCR / 16S rRNA / MALDI-TOFSource:
Acta Veterinaria-Beograd, 2016, 66, 3, 304-316Publisher:
- Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
Funding / projects:
- Implementation and evaluation of a new molecular method for a quick detection of a mecA gene directly in swabs originated from humans, animals and their environment (RS-MESTD-Technological Development (TD or TR)-31079)
DOI: 10.1515/acve-2016-0027
ISSN: 0567-8315
WoS: 000384771100003
Scopus: 2-s2.0-84992223383
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Institution/Community
Fakultet veterinarske medicineTY - JOUR AU - Aksentijević, Ksenija AU - Ašanin, Jelena AU - Milivojević, Dušan AU - Čolović, Svetlana AU - Butorac, Ana AU - Cindrić, Mario AU - Mišić, Dušan PY - 2016 UR - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1360 AB - 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 identification of the 51 isolates originated from fish, 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 confirmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confirmed 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 identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Although modern test methods that have very high specificity (PCR, 16S rRNA gene Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods. AB - S obzirom na značaj stečene rezistencije Pseudomonas vrsta, kao i na intrinzičnu rezistenciju Stenotrophomonas vrsta, a u cilju preciznog ispitivanja osetljivosti na antibiotike, neophodna je jasna diferencijacija pripadnika ovih rodova bakterija. U tom cilju su u identifikaciji 51 izolata poreklom od riba korišćene metode: standardni PCR, 16S rRNA sekvenciranje gena, MALDI-TOF. Rezultati standardnog PCR testa, 16S rRNA sekvenciranja gena i MALDI-TOF analize su za 35 sojeva potvrdili pripadnost rodu Pseudomonas. Standardnim PCR testom i primenom aparata VITEK MS utvrđeno je da 10 sojeva pripada vrsti Stenotrophomonas maltophilia. U 16S rRNA sekvenciranju gena 3 soja koja su bila pozitivna u oba standardna PCR testa identifikovana su kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. VITEK MS je ova tri soja u prvoj identifikaciji identifikovao kao 99% Stenotrophomonas, a u ponovljenoj identifikaciji kao 99% Pseudomonas. Ti sojevi su na aparatu MALDI TOF/TOF 4800 Plus bili identifikovani kao Stenotrophomonas. U 16S rRNA sekvenciranju gena tri soja koja su bila negativna u oba standardna PCR testa su identifikovana kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. Aparat VITEK MS je ova tri soja identifikovao u jednoj identifikaciji kao 99% Stenotrophomonas, a u drugoj identifikaciji 99% kao Pseudomonas. Ti sojevi su na aparatu MALDI TOF/ TOF 4800 Plus bili identifikovani kao Stenotrophomonas. Iako su u ovom istraživanju korišćene savremene metode ispitivanja koje imaju vrlo visoku specifičnost (PCR, 16s rRNK sequencing, MALDI TOF) precizna diferencijacija Pseudomonas i Stenotrophomonas vrsta nije mogla biti postignuta. PB - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd T2 - Acta Veterinaria-Beograd T1 - Differentiation between Pseudomonas and Stenotrophomonas species isolated from fish using molecular and MALDI-TOF method T1 - Diferencijacija Pseudomonas i Stenotrophomonas vrsta izolovanih iz riba primenom molekularnih metoda i MALDI-TOF metode VL - 66 IS - 3 SP - 304 EP - 316 DO - 10.1515/acve-2016-0027 ER -
@article{ author = "Aksentijević, Ksenija and Ašanin, Jelena and Milivojević, Dušan and Čolović, Svetlana and Butorac, Ana and Cindrić, Mario and Mišić, Dušan", year = "2016", abstract = "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 identification of the 51 isolates originated from fish, 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 confirmed 35 strains to belong to the Pseudomonas genus. Standard PCR test and VITEK MS device confirmed 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 identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Three strains were negative in both standard PCR tests for Pseudomonas and Stenotrpohomonas. 16S rRNA gene sequencing identified these 3 strains to be 99% Pseudomonas sp. and 99% Stenotrophomonas sp. VITEK MS first identified these three strains as 99% Stenotrophomonas, and in the repeated identification it identified them as 99% Pseudomonas. MALDI TOF/TOF 4800 Plus device identified these strains as Stenotrophomonas. Although modern test methods that have very high specificity (PCR, 16S rRNA gene Pseudomonas and Stenotrophomonas species for 6 isolates could not be reached using the above mentioned methods., S obzirom na značaj stečene rezistencije Pseudomonas vrsta, kao i na intrinzičnu rezistenciju Stenotrophomonas vrsta, a u cilju preciznog ispitivanja osetljivosti na antibiotike, neophodna je jasna diferencijacija pripadnika ovih rodova bakterija. U tom cilju su u identifikaciji 51 izolata poreklom od riba korišćene metode: standardni PCR, 16S rRNA sekvenciranje gena, MALDI-TOF. Rezultati standardnog PCR testa, 16S rRNA sekvenciranja gena i MALDI-TOF analize su za 35 sojeva potvrdili pripadnost rodu Pseudomonas. Standardnim PCR testom i primenom aparata VITEK MS utvrđeno je da 10 sojeva pripada vrsti Stenotrophomonas maltophilia. U 16S rRNA sekvenciranju gena 3 soja koja su bila pozitivna u oba standardna PCR testa identifikovana su kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. VITEK MS je ova tri soja u prvoj identifikaciji identifikovao kao 99% Stenotrophomonas, a u ponovljenoj identifikaciji kao 99% Pseudomonas. Ti sojevi su na aparatu MALDI TOF/TOF 4800 Plus bili identifikovani kao Stenotrophomonas. U 16S rRNA sekvenciranju gena tri soja koja su bila negativna u oba standardna PCR testa su identifikovana kao 99% Pseudomonas sp. i 99% Stenotrophomonas sp. Aparat VITEK MS je ova tri soja identifikovao u jednoj identifikaciji kao 99% Stenotrophomonas, a u drugoj identifikaciji 99% kao Pseudomonas. Ti sojevi su na aparatu MALDI TOF/ TOF 4800 Plus bili identifikovani kao Stenotrophomonas. Iako su u ovom istraživanju korišćene savremene metode ispitivanja koje imaju vrlo visoku specifičnost (PCR, 16s rRNK sequencing, MALDI TOF) precizna diferencijacija Pseudomonas i Stenotrophomonas vrsta nije mogla biti postignuta.", publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd", journal = "Acta Veterinaria-Beograd", title = "Differentiation between Pseudomonas and Stenotrophomonas species isolated from fish using molecular and MALDI-TOF method, Diferencijacija Pseudomonas i Stenotrophomonas vrsta izolovanih iz riba primenom molekularnih metoda i MALDI-TOF metode", volume = "66", number = "3", pages = "304-316", doi = "10.1515/acve-2016-0027" }
Aksentijević, K., Ašanin, J., Milivojević, D., Čolović, S., Butorac, A., Cindrić, M.,& Mišić, D.. (2016). Differentiation between Pseudomonas and Stenotrophomonas species isolated from fish using molecular and MALDI-TOF method. in Acta Veterinaria-Beograd Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 66(3), 304-316. https://doi.org/10.1515/acve-2016-0027
Aksentijević K, Ašanin J, Milivojević D, Čolović S, Butorac A, Cindrić M, Mišić D. Differentiation between Pseudomonas and Stenotrophomonas species isolated from fish using molecular and MALDI-TOF method. in Acta Veterinaria-Beograd. 2016;66(3):304-316. doi:10.1515/acve-2016-0027 .
Aksentijević, Ksenija, Ašanin, Jelena, Milivojević, Dušan, Čolović, Svetlana, Butorac, Ana, Cindrić, Mario, Mišić, Dušan, "Differentiation between Pseudomonas and Stenotrophomonas species isolated from fish using molecular and MALDI-TOF method" in Acta Veterinaria-Beograd, 66, no. 3 (2016):304-316, https://doi.org/10.1515/acve-2016-0027 . .