| Literature DB >> 30207519 |
Victoria Stadler Tasca Ribeiro1, Felipe Francisco Tuon2,1, Letícia Kraft1, Paula Hansen Suss1, Luciana Cristina Wollmann1, João Gabriel Roderjan1, Diego Armando Brito3, Fabiana Alexandrino4, Juliane Soldi Malgarin4, Luis Gustavo Morello4,5, Francisco Diniz Affonso da Costa1,2, Marcelo Pillonetto2,3.
Abstract
Real-time polymerase chain reaction (qPCR) using 16S rDNA is an alternative to conventional culture-based tests. The aim of this study was to compare the conventional culture method with qPCR using 16S rDNA in a model of cardiac tissue contamination. Samples of cardiac tissue for artificial contamination with Escherichia coli and control samples were submitted for DNA extraction, which was conducted by selective and alkaline lysis and purification steps. A standard curve for 16S rDNA was constructed to determine the efficiency and analytical sensitivity of the assay in concentrations from 106 to 102 c.f.u. ml-1 using TaqMan Master Mix. 16S rDNA was detected in all contaminated samples; however, it was not detected in the the final washing step solution of the sample with a bioburden of 102 c.f.u. ml-1. Using qPCR is a potential alternative to conventional culture for microbiological safety testing of allograft tissues for biobanking, reducing the time and labour input required.Entities:
Keywords: 16S rDNA; cardiovascular tissues; conventional culture; real-time PCR; tissue bank
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Year: 2018 PMID： 30207519 DOI： 10.1099/jmm.0.000837
Source DB: PubMed Journal: J Med Microbiol ISSN： 0022-2615 Impact factor: 2.472