INTRODUCTION: Nowadays, on-a-chip capillary electrophoresis is a routine method for the detection of PCR fragments. The Agilent 2100 Bioanalyzer was one of the first commercial devices in this field. Our project was designed to study the characteristics of Agilent DNA 1000 kit in PCR fragment analysis as a part of circulating tumour cell (CTC) detection technique. Despite the common use of this kit a complex analysis of the results from a long-term project is still missing. MATERIALS AND METHODS: A commercially available Agilent DNA 1000 kit was used as a final step in the CTC detection (AdnaTest) for the determination of the presence of PCR fragments generated by Multiplex PCR. Data from 30 prostate cancer patients obtained during two years of research were analyzed to determine the trueness and precision of the PCR fragment size determination. Additional experiments were performed to demonstrate the precision (repeatability, reproducibility) and robustness of PCR fragment concentration determination. RESULTS: The trueness and precision of the size determination was below 3% and 2% respectively. The repeatability of the concentration determination was below 15%. The difference in concentration determination increases when Multiplex-PCR/storage step is added between the two measurements of one sample. CONCLUSIONS: The characteristics established in our study are in concordance with the manufacturer's specifications established for a ladder as a sample. However, the concentration determination may vary depending on chip preparation, sample storage and concentration. The 15% variation of concentration determination repeatability was shown to be partly proportional and can be suppressed by proper normalization.

• Klíčová slova
• Agilent DNA 1000 kit, capillary electrophoresis, circulating tumour cells, lab-on-a-chip devices, multiplex PCR,
• MeSH
• aktiny genetika   MeSH
• antigeny povrchové genetika   MeSH
• DNA nádorová genetika   MeSH
• erbB receptory genetika   MeSH
• glutamátkarboxypeptidasa II genetika   MeSH
• laboratoř na čipu * MeSH
• lidé MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• nádorové biomarkery genetika   MeSH
• nádorové cirkulující buňky metabolismus   MeSH
• nádory prostaty rezistentní na kastraci krev   genetika   MeSH
• prostatický specifický antigen genetika   MeSH
• reagenční diagnostické soupravy MeSH
• regulace genové exprese u nádorů * MeSH
• reprodukovatelnost výsledků MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• antigeny povrchové MeSH
• DNA nádorová MeSH
• erbB receptory MeSH
• FOLH1 protein, human MeSH Prohlížeč
• glutamátkarboxypeptidasa II MeSH
• nádorové biomarkery MeSH
• prostatický specifický antigen MeSH
• reagenční diagnostické soupravy MeSH