Aeromonas detection and characterization using genus-specific PCR and single-strand conformation polymorphism (SSCP)
2012
Delamare, Ana Paula Longaray | Lucena, Roberto Francisco | Thomazi, Guilherme | Ferrarini, Shana | Zacaria, Jucimar | Echeverrigaray, Sergio
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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