Design of a Cell-Free DNA Isolation method from maternal plasma based on Metal Organic Frameworks; with the aim of use in prenatal sex determination
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Mohammad Mahdi Abolhosseini,1 Banafsheh Rastegari,2,* Rita Arabsolghar,3 Abbas Behzad Behbahani,4
1. Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
2. Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
3. Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
4. Diagnostic Laboratory Sciences and Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
- Introduction: Many sex-linked genetic diseases are identified in humans, with male fetuses more exposed to the complications of the diseases. So, prenatal sexing seems important in identifying these infants. Common methods are unable to define fetal gender safely, early and accurately. The use of cell-free fetal DNA has been in the focus of researchers. One of the challenges of using this method is the isolation of fetal DNA from maternal plasma. Commercial kits are expensive and offering low yield of DNA. In this project, we intend to isolate fetal DNA from maternal plasma using metal organic frameworks nanoparticles labeled with specific probes of the SRY gene specific for the Y chromosome.
- Methods: Y-Chromosome Specific probes were immobilized on BIOMIL-4 metal organic frameworks (M.O.Fs) and the resulting construct was characterised using XRD and FTIR methods. These functionalized M.O.Fs were added to maternal plasma aliquots for the target DNA fragments to hybridize to the probes. Conventional and Real-Time Polymerase Chain Reaction were performed to confirm isolation of target DNA fragments.
- Results: The XRD curve confirmed that immobilization of dna probes on MOFs did not alter the crystaline structure of the frameworks. The FTIR analysis also confirmed the presence of chemical bonds reflecting the presence of nucleic acids in the surface of MOFs. The PCR reaction successfully amplified targeted DNA fragments, confirming the isolation of Y chromosome specific DNA.
- Conclusion: Using metal organic frameworks for immobilization of hybridization probes may be a method of interest for isolation of specific DNA fragments from clinical samples.
- Keywords: Cell-free fetal DNA, Metal-Organic Frameworks, Hybridization Probes, Sex determination
