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Enhanced Genetic Analysis of Single Human Bioparticles Recovered by Simplified Micromanipulation from Forensic ‘Touch DNA’ Evidence
Published on: March 9, 2015
A selected pre-amplification strategy for genetic analysis using limited DNA targets
Peng Xia1, Ramin Radpour, Corina Kohler
1Women's Hospital, Department of Biomedicine, University of Basel, Basel, Switzerland.
Clinical Chemistry and Laboratory Medicine
|August 14, 2009
Summary
A new pre-amplification strategy dramatically increases fetal DNA in maternal plasma, improving genetic testing with limited DNA. This method enhances minor target detection but is not suitable for precise quantification.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Limited DNA resources or targets in genetic testing can cause misdiagnosis.
- A novel strategy was developed to selectively amplify minor DNA targets.
Purpose of the Study:
- To evaluate a selective pre-amplification strategy using cell-free (ccf) male fetal DNA in maternal plasma.
- To assess the efficiency of increasing minor target DNA in a predominant background.
Main Methods:
- Utilized circulating cell-free (ccf) male fetal DNA as a minor target in maternal plasma.
- Employed Expand Long Template PCR for selective pre-amplification of the SRY locus (Y chromosome).
- Assessed amplification efficiency using real-time PCR for SRY and GAPDH (total DNA).
Main Results:
- Selective pre-amplification significantly increased ccf fetal DNA (p=0.000, fold change=11,596).
- The proportion of ccf fetal DNA rose from 2.19% to 25,334% post-amplification.
- Increased fetal DNA amounts were uncorrelated with pre-amplification levels (r=-0.017, p=0.949).
Conclusions:
- The strategy shows potential for genetic analysis with limited DNA resources and targets.
- Quantitative assessment is not recommended due to observed imbalanced amplification.
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