Contribution of PCR Denaturing Gradient Gel Electrophoresis Combined with Mixed Chromatogram Software Separation for Complex Urinary Sample Analysis

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Authors

KOTÁSKOVÁ Iva MALIŠOVÁ Barbora OBRUČOVÁ Hana HOLÁ Veronika PEROUTKOVÁ Tereza RŮŽIČKA Filip FREIBERGER Tomáš

Year of publication 2017
Type Article in Periodical
Magazine / Source Journal of Molecular Microbiology and Biotechnology
MU Faculty or unit

Faculty of Medicine

Citation
Doi http://dx.doi.org/10.1159/000484524
Field Genetics and molecular biology
Keywords Catheter; Complex sample; Mixed chromatogram; PCR denaturing gradient gel electrophoresis; RipSeq Mixed
Description Complex samples are a challenge for sequencing-based broad-range diagnostics. We analysed 19 urinary catheter, ureteral Double-J catheter, and urine samples using 3 methodological approaches. Out of the total 84 operational taxonomic units, 37, 61, and 88% were identified by culture, PCR-DGGE-SS (PCR denaturing gradient gel electrophoresis followed by Sanger sequencing), and PCR-DGGE-RM (PCR- DGGE combined with software chromatogram separation by RipSeq Mixed tool), respectively. The latter approach was shown to be an efficient tool to complement culture in complex sample assessment.
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