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Aeromonas detection and characterization using genus-specific PCR and single-strand conformation polymorphism (SSCP)

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Abstract

Based on sequence alignment, oligonucleotide primers targeting the Aeromonas extracellular lipase gene were developed for PCR detection of member of the genus. A pair of primers designed for conserved regions of the gene amplified a 276 bp sequence in all Aeromonas species and tested strains, but did not have a positive result with other Gram-positive and Gram-negative bacteria, showing high specificity and sensitivity. Selective enrichment in alkaline peptone water, followed by centrifugation, and direct usage of cells suspension as template, detected initial populations of 10 c.f.u. ml−1. Single-strand conformation polymorphism analysis of the PCR products allowed the characterization of Aeromonas strains with a high discriminatory power (Simpson′s index = 0.988). The method presented here provides a useful tool for the rapid detection of Aeromonas and the characterization of Aeromonas isolates.

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Acknowledgments

We are grateful to the University of Caxias do Sul for the financial support, to CAPES, and CNPq for scholarship funds.

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Authors and Affiliations

  1. Institute of Biotechnology, University of Caxias do Sul, R. Francisco G. Vargas 1130, Caxias do Sul, Rio Grande do Sul, 95001-970, Brazil

    Ana Paula Longaray Delamare, Roberto Francisco Lucena, Guilherme Thomazi, Shana Ferrarini, Jucimar Zacaria & Sergio Echeverrigaray

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  1. Ana Paula Longaray Delamare
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  2. Roberto Francisco Lucena
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  3. Guilherme Thomazi
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  4. Shana Ferrarini
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  5. Jucimar Zacaria
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  6. Sergio Echeverrigaray
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Correspondence to Ana Paula Longaray Delamare.

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Delamare, A.P.L., Lucena, R.F., Thomazi, G. et al. Aeromonas detection and characterization using genus-specific PCR and single-strand conformation polymorphism (SSCP). World J Microbiol Biotechnol 28, 3007–3013 (2012). https://doi.org/10.1007/s11274-012-1111-5

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  • DOI: https://doi.org/10.1007/s11274-012-1111-5

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