Conventional culture method and qPCR using 16S rDNA for tissue bank: a comparison using a model of cardiac tissue contamination

• DOI: 10.1099/jmm.0.000837
• Publication Date: 2018-09-12T15:12:49Z
• Authors: Victoria Stadler ...; Felipe Francisco ...; Letícia Kraft; Paula Hansen Suss; Luciana Cristina ...; João Gabriel Rode...; Diego Armando Brito; Fabiana Alexandrino; Juliane Soldi Mal...; Luis Gustavo Morello; Francisco Diniz A...; Marcelo Pillonetto
• Keywords: DNA, Bacterial; 0301 basic medicine; Bacteriological Techniques; Myocardium; Heart; Bacterial Infections; Tissue Banks; Real-Time Polymerase Chain Reaction; DNA, Ribosomal; Sensitivity and Specificity; 3. Good health; 03 medical and health sciences; RNA, Ribosomal, 16S; Escherichia coli; Heart Transplantation; Humans

Real-time polymerase chain reaction (qPCR) using 16S rDNA is an alternative to conventional culture-based tests. The aim of this study was compare the culture method with qPCR in a model cardiac tissue contamination. Samples for artificial contamination Escherichia coli and control samples were submitted DNA extraction, which conducted by selective alkaline lysis purification steps. A standard curve constructed determine efficiency analytical sensitivity assay concentrations from 106 102 c.f.u. ml-1 TaqMan Master Mix. detected all contaminated samples; however, it not final washing step solution sample bioburden ml-1. Using potential microbiological safety testing allograft tissues biobanking, reducing time labour input required.