Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it>
<p>Abstract</p> <p>Background</p> <p>We assessed the potential of multilocus variable-number tandem-repeat analysis (MLVA), pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing for discriminating 104 sporadic and outbreak-related <it>Y...
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| Format: | Article |
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BMC
2011-02-01
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| Series: | BMC Microbiology |
| Online Access: | http://www.biomedcentral.com/1471-2180/11/42 |
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| author | Skurnik Mikael Kuusi Markku Haukka Kaisa Toivonen Susanna Sihvonen Leila M Siitonen Anja |
| author_facet | Skurnik Mikael Kuusi Markku Haukka Kaisa Toivonen Susanna Sihvonen Leila M Siitonen Anja |
| author_sort | Skurnik Mikael |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>We assessed the potential of multilocus variable-number tandem-repeat analysis (MLVA), pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing for discriminating 104 sporadic and outbreak-related <it>Yersinia enterocolitica </it>(YE) bio/serotype 3-4/O:3 and 2/O:9 isolates. MLVA using six VNTR markers was performed in two separate multiplex PCRs, and the fluorescently labeled PCR products were accurately sized on an automated DNA sequencer.</p> <p>Results</p> <p>MLVA discriminated 82 sporadic YE 3-4/O:3 and 2/O:9 strains into 77 types, whereas PFGE with the restriction enzyme <it>Not</it>I discriminated the strains into 23 different PFGE pulsotypes. The discriminatory index for a sporadic strain was 0.862 for PFGE and 0.999 for MLVA. MLVA confirmed that a foodborne outbreak in the city of Kotka, Finland in 2003 had been caused by a multiresistant YE 4/O:3 strain that was distinctly different from those of epidemiologically unrelated strains with an identical PFGE pulsotype. The multiresistance of <it>Y. enterocolitica </it>strains (19% of the sporadic strains) correlated significantly (p = 0.002) with travel abroad. All of the multiresistant <it>Y. enterocolitica </it>strains belonged to four PFGE pulsotypes that did not contain any susceptible strains. Resistance to nalidixic acid was related to changes in codons 83 or 87 that stemmed from mutations in the <it>gyrA </it>gene. The conjugation experiments demonstrated that resistance to CHL, STR, and SUL was carried by a conjugative plasmid.</p> <p>Conclusions</p> <p>MLVA using six loci had better discriminatory power than PFGE with the <it>Not</it>I enzyme. MLVA was also a less labor-intensive method than PFGE and the results were easier to analyze. The conjugation experiments demonstrated that a resistance plasmid can easily be transferred between <it>Y. enterocolitica </it>strains. Antimicrobial multiresistance of <it>Y. enterocolitica </it>strains was significantly associated with travel abroad.</p> |
| first_indexed | 2024-12-12T22:50:58Z |
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| id | doaj.art-0d4a185f0bb9432faeffa7f024c0cf57 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
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| publishDate | 2011-02-01 |
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| spelling | doaj.art-0d4a185f0bb9432faeffa7f024c0cf572022-12-22T00:09:04ZengBMCBMC Microbiology1471-21802011-02-011114210.1186/1471-2180-11-42Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it>Skurnik MikaelKuusi MarkkuHaukka KaisaToivonen SusannaSihvonen Leila MSiitonen Anja<p>Abstract</p> <p>Background</p> <p>We assessed the potential of multilocus variable-number tandem-repeat analysis (MLVA), pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing for discriminating 104 sporadic and outbreak-related <it>Yersinia enterocolitica </it>(YE) bio/serotype 3-4/O:3 and 2/O:9 isolates. MLVA using six VNTR markers was performed in two separate multiplex PCRs, and the fluorescently labeled PCR products were accurately sized on an automated DNA sequencer.</p> <p>Results</p> <p>MLVA discriminated 82 sporadic YE 3-4/O:3 and 2/O:9 strains into 77 types, whereas PFGE with the restriction enzyme <it>Not</it>I discriminated the strains into 23 different PFGE pulsotypes. The discriminatory index for a sporadic strain was 0.862 for PFGE and 0.999 for MLVA. MLVA confirmed that a foodborne outbreak in the city of Kotka, Finland in 2003 had been caused by a multiresistant YE 4/O:3 strain that was distinctly different from those of epidemiologically unrelated strains with an identical PFGE pulsotype. The multiresistance of <it>Y. enterocolitica </it>strains (19% of the sporadic strains) correlated significantly (p = 0.002) with travel abroad. All of the multiresistant <it>Y. enterocolitica </it>strains belonged to four PFGE pulsotypes that did not contain any susceptible strains. Resistance to nalidixic acid was related to changes in codons 83 or 87 that stemmed from mutations in the <it>gyrA </it>gene. The conjugation experiments demonstrated that resistance to CHL, STR, and SUL was carried by a conjugative plasmid.</p> <p>Conclusions</p> <p>MLVA using six loci had better discriminatory power than PFGE with the <it>Not</it>I enzyme. MLVA was also a less labor-intensive method than PFGE and the results were easier to analyze. The conjugation experiments demonstrated that a resistance plasmid can easily be transferred between <it>Y. enterocolitica </it>strains. Antimicrobial multiresistance of <it>Y. enterocolitica </it>strains was significantly associated with travel abroad.</p>http://www.biomedcentral.com/1471-2180/11/42 |
| spellingShingle | Skurnik Mikael Kuusi Markku Haukka Kaisa Toivonen Susanna Sihvonen Leila M Siitonen Anja Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it> BMC Microbiology |
| title | Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it> |
| title_full | Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it> |
| title_fullStr | Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it> |
| title_full_unstemmed | Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it> |
| title_short | Multilocus Variable-Number Tandem-Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Patterns in Discrimination of Sporadic and Outbreak-Related Strains of <it>Yersinia enterocolitica</it> |
| title_sort | multilocus variable number tandem repeat analysis pulsed field gel electrophoresis and antimicrobial susceptibility patterns in discrimination of sporadic and outbreak related strains of it yersinia enterocolitica it |
| url | http://www.biomedcentral.com/1471-2180/11/42 |
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