Author/Authors :
Keshavarz, Zeinab Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences , Moezzi, Leili Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences , Ranjbaran, Reza Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences , Aboualizadeh, Farzaneh Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences , Behzad-Behbahani, Abbas Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences , Abdullahi, Masooma Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences , Sharifzadeh, Sedigheh Diagnostic Laboratory Sciences and Technology Research Center - School of Paramedical Sciences - Shiraz University of Medical Sciences
Abstract :
Background: Discovery of short cell free fetal DNA (cffDNA) fragments in maternal
plasma has created major changes in the field of prenatal diagnosis. The use of
cffDNA to set up noninvasive prenatal test is limited due to the low concentration of
fetal DNA in maternal plasma therefore, employing a high efficiency extraction
method leads to more accurate results. The aim of this study was to evaluate the efficiency
of Triton/Heat/Phenol (THP) protocol in comparison with the QIAamp DNA
Blood mini Kit for cffDNA purification.
Methods: In order to evaluate the efficiency of THP protocol, DNA of Rhesus D (RhD)
negative pregnant women's plasma was collected, then real-time PCR for RHD exon
7 was performed. The Ct value data of real time PCR obtained by two different
methods were compared and after delivery serology test on cord blood was done to
validate the real time PCR results.
Results: The results indicated significant differences between two extraction methods
(p=0.001). The mean±SD of Ct-value using THP protocol was 33.8±1.6 and 36.1±2.47
using QIAamp DNA Blood mini Kit.
Conclusion: our finding demonstrated that THP protocol was more effective than the
QIAamp DNA Blood mini Kits for cffDNA extraction and lead to decrease the false
negative results.
Keywords :
THP protocol , Real time polymerase chain reaction , Prenatal diagnosis , Fetus
