RHD Gene Amplification on Reused FTA Card: A Pilot Study
Published: 2022-03-21
Page: 72-75
Issue: 2022 - Volume 5 [Issue 2]
Mujtaba A. Allawati *
Department of Medical Laboratory Science, Oman College of Health Sciences, Oman.
*Author to whom correspondence should be addressed.
Abstract
Aims: RHD gene analysis performed directly from repeatedly extracted DNA on FTA card disc has not been studied. Here we demonstrate a direct real-time PCR of RHD gene exon 10 by reusing FTA card.
Study Design: This is an experimental pilot study to observe the RHD exon 10 amplification on repeatedly extracted DNA on FTA card.
Methodology: Five samples from dry blood spot on FTA cards were subjected to wash and lysis four times without additional extraction process. The lysates on each round was demonstrated for signs of amplification by melting curve analysis and agarose gel electrophoresis.
Results: Successful amplification of RHD gene exon 10 was observed by melting curve analysis (melting temperature 84.85oC) and reconfirmed by 2% agarose gel electrophoresis (393 bp fragment).
Conclusion: Repeated amplification of DNA from direct FTA card with dried blood sample without the need of time consuming extraction process was possible.
Keywords: FTA card, RHD gene, real-time PCR