Evaluation Of DNA Extraction From Archieved Formalin-Fixed Paraffin Tissues (FFPT) By β-Globin Gene PCR.

Abstract

Background: the knowledge of the nucleic acid state before start to molecular detection have a great importance to make a math measurements by which the nucleic acid quantity and quality will be knows.

Objective: Conventional methods have limitations for extracting amplifiable DNA from specimens containing a small number of cells. DNA quality and quantity was evaluated by β-globin amplification and Nanodrop instrument respectively. DNA extraction from FFPT has become essential for molecular research specially with advances in molecular technology.

Methodology: The current study tested breast cancer FFPT specimens of Iraqi women which collected from AL-Sader teaching hospital histopathology and neoplasm units , and several privet histopathology labs in AL-Najaf province, the specimens were stored for the years (2008,2009,2010 and 2011). The specimens were included of BC 54 , fibroadenoma 21, mastitis 18, and ductectesia 19 samples.

Results: Specific primers were used to detect the activity of beta-globin gene by PCR technique and the results appeared that The mean of DNA quantity was (61.68 ng/ul) while the DNA extract quality which appeared positive by β-glubine PCR was 71.42% (80 of 112), BC 70.37% (38 of 54), fibroadenoma 85.71% (18 of 21), mastitis 77.77% (14 of 18) and ductectesia 52.63 % (10 of 19), there was a significant difference ( p<0.5). The beta-globin PCR appeared different results depending on the DNA stats.

Conclusions: the evaluation of the extracted DNA by Nanodrop system and beta-globin gene is the easiest and definite method compared with other researchers who worked of other evaluation methods.

Recommendations: This study recommends that every study performed on DNA should be started with beta-globin testing to avoid false negative results caused by degraded DNA.