Non-invasive prenatal detection of trisomy 21 using tandem single nucleotide polymorphisms
Sujana Ghanta, Michael E Mitchell, Mary Ames, Mats Hidestrand, Pippa Simpson, Mary Goetsch, William G Thilly, Craig A Struble, Aoy Tomita-Mitchell, Sujana Ghanta, Michael E Mitchell, Mary Ames, Mats Hidestrand, Pippa Simpson, Mary Goetsch, William G Thilly, Craig A Struble, Aoy Tomita-Mitchell
Abstract
Background: Screening tests for Trisomy 21 (T21), also known as Down syndrome, are routinely performed for the majority of pregnant women. However, current tests rely on either evaluating non-specific markers, which lead to false negative and false positive results, or on invasive tests, which while highly accurate, are expensive and carry a risk of fetal loss. We outline a novel, rapid, highly sensitive, and targeted approach to non-invasively detect fetal T21 using maternal plasma DNA.
Methods and findings: Highly heterozygous tandem Single Nucleotide Polymorphism (SNP) sequences on chromosome 21 were analyzed using High-Fidelity PCR and Cycling Temperature Capillary Electrophoresis (CTCE). This approach was used to blindly analyze plasma DNA obtained from peripheral blood from 40 high risk pregnant women, in adherence to a Medical College of Wisconsin Institutional Review Board approved protocol. Tandem SNP sequences were informative when the mother was heterozygous and a third paternal haplotype was present, permitting a quantitative comparison between the maternally inherited haplotype and the paternally inherited haplotype to infer fetal chromosomal dosage by calculating a Haplotype Ratio (HR). 27 subjects were assessable; 13 subjects were not informative due to either low DNA yield or were not informative at the tandem SNP sequences examined. All results were confirmed by a procedure (amniocentesis/CVS) or at postnatal follow-up. Twenty subjects were identified as carrying a disomy 21 fetus (with two copies of chromosome 21) and seven subjects were identified as carrying a T21 fetus. The sensitivity and the specificity of the assay was 100% when HR values lying between 3/5 and 5/3 were used as a threshold for normal subjects.
Conclusions: In summary, a targeted approach, based on calculation of Haplotype Ratios from tandem SNP sequences combined with a sensitive and quantitative DNA measurement technology can be used to accurately detect fetal T21 in maternal plasma when sufficient fetal DNA is present in maternal plasma.
Conflict of interest statement
Competing Interests: Sujana Ghanta, Mary Ames, Mats Hidestrand, Mary Goetsch, and Craig Struble have been given stock options of Tandem Diagnostics, the company that is developing the technology the authors have reported in this study. Sujana Ghanta, Mary Ames, and Craig Struble currently serve as consultants to Tandem Diagnostics. Michael E. Mitchell is a board member, shareholder, and consultant of Tandem Diagnostics. Aoy Tomita-Mitchell is a shareholder of Tandem Diagnostics. Tandem Diagnostics also currently has a sponsored research with Dr. Aoy Tomita-Mitchell's and Michael E. Mitchell's labs at the Medical College of Wisconsin for research that is further development of this reported technology. Mats Hidestrand and Mary Goetsch work for Dr. Aoy Tomita-Mitchell's lab at the Medical College of Wisconsin and receive part of their salaries for this research. Michael E. Mitchell, William Thilly, and Aoy Tomita-Mitchell are also inventors on patent applications involving the technology reported herein. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials.
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