Transrenal DNA as a diagnostic tool: important technical notes

Annals of the New York Academy of Sciences
Ying-Hsiu SuSamuil Umansky

Abstract

A small portion of DNA from apoptotic cells escapes complete degradation, appears in blood as oligonucleosomal-size fragments, is excreted in the urine, and can be used for diagnostic purposes. More detailed study revealed that transrenal DNA (Tr-DNA) belongs to a relatively low molecular-weight (150-250 bp) fraction, thereby requiring more careful attention to methods employed for purification and analysis. For example, here it is demonstrated that the QIAamp blood kit purifies primarily high molecular-weight DNA from serum, whereas the Guanidine/Promega Wizard Resin (GITC/WR) method purifies primarily low molecular-weight DNA. As a result, sensitivity in detection of K-RAS mutations in serum of patients with colorectal tumors is significantly higher with DNA isolated with the GITC/WR method than with the QIAamp kit. Amplicon size is also extremely important in analysis of Tr-DNA, because the shorter the amplicon, the higher is the sensitivity of biomarker detection in Tr-DNA. One hundred fifty-seven and 87 bp amplicons were employed for detection of mutant K-RAS in DNA isolated from 0.1 mL of urine obtained from 15 patients with pancreatic cancer. Mutant K-RAS was found in Tr-DNA of 3 and 10 patients with the long and short a...Continue Reading

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Citations

May 19, 2010·Biomarkers in Medicine·Barbara L Parsons, Fanxue Meng
Jun 7, 2014·Critical Care : the Official Journal of the Critical Care Forum·Jose Garnacho-MonteroAntonio Puppo-Moreno
Nov 11, 2014·Hepatology Research : the Official Journal of the Japan Society of Hepatology·Surbhi JainYing-Hsiu Su
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Dec 2, 2006·Biochimica Et Biophysica Acta·M Fleischhacker, B Schmidt
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