Comparisons between capillary zone electrophoresis and real-time PCR for quantification of circulating DNA levels in human sera.
BACKGROUND
Recently, some research results showed that the circulating DNA in serum or plasma had potential for the molecular diagnosis and prognosis of certain cancers. Several methods have been employed for the quantification of circulating DNA. However, the circulating DNA levels obtained by various methods exhibited considerable differences. Additionally, these methods were labor-extensive and time-consuming, and not suitable for the quantification of circulating DNA in numerous samples due to the use of commercial DNA extraction kits for the purification of circulating DNA. We presented a new method for the quantification of circulating DNA in sera by capillary zone electrophoresis (CZE) with laser-induced fluorescence detection (LIF).


METHODS
In the present work, we want to make comparison between CZE-LIF assay and real time PCR for the quantification of circulating DNA levels. Linearity, intra and inter variability of two methods were evaluated.


RESULTS
The intra and inter variability of circulating DNA quantification by real-time PCR were 7.3% and 14.92%, respectively. In CZE assay the intra and inter variability were 4.19% and 6.91%, respectively. The R.S.D. values of the same coated capillary and different coated capillaries were 5.14% and 9.02%, respectively. Our data showed that the circulating DNA levels obtained by two methods had a good correlation. Moreover, we further confirmed that blood samples collection, serum preparation and other treatment procedures had a significant impact on the DNA levels in sera.


CONCLUSIONS
Our data further illustrated that CZE-LIF is a simple, rapid and sensitive method for the quantification of circulating DNA in human sera, and well suitable for the analysis of a large number of samples in clinical diagnosis.
Sang
Fuming
F
Department of Chemistry, Shanghai Jiaotong University, 800 Dongchuan Road, Shanghai 200240, PR China.
Ren
Jicun
J
D002138:Calibration; D004247:DNA / Q000097:blood*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D044967:Serum / Q000737:chemistry*
D004247:DNA