Quantity versus quality: Optimal methods for cell-free DNA isolation from plasma of pregnant women Article

Jorgez, CJ, Dang, DD, Simpson, JL et al. (2006). Quantity versus quality: Optimal methods for cell-free DNA isolation from plasma of pregnant women . 8(10), 615-619. 10.1097/01.gim.0000241904.32039.6f

cited authors

  • Jorgez, CJ; Dang, DD; Simpson, JL; Lewis, DE; Bischoff, FZ

fiu authors

abstract

  • PURPOSE: Methods to isolate cell-free fetal DNA from maternal plasma are critical in developing noninvasive fetal DNA testing strategies. Given that plasma consists of heterogeneous DNA-size fragments in a complex mix of proteins, recovery and analysis of this DNA are understandably inefficient. To facilitate recovery, we performed qualitative and quantitative analysis of DNA isolated from maternal plasma. METHODS: DNA isolated from maternal blood (n = 15) was compared using five different DNA isolation protocols: two conventional, two column-based, and one magnetic-bead based. Purity and concentration of DNA recovered were determined with a NanoDrop spectrophotometer. Real-time polymerase chain reaction quantification of the β-globin and DYS1 loci was performed to determine total and fetal-specific genome equivalents, respectively. RESULTS: DNA quality and quantity were different among the five methods tested. Although purity and concentration of total DNA were greatest with the conventional boiling-lysis approach, correct detection of a male fetus was achieved in only 62.5% of cases. DNA isolation using the magnetic beads yielded the highest quantity of total DNA (2018.83 ± 4.09 GEq/mL), with 100% fetal DNA detection. CONCLUSIONS: Optimal plasma DNA recovery protocols must take into account DNA purity and concentration. We confirm that the magnetic-beads method provides a fast, simple, sensitive, and specific approach to purify plasma DNA. The resulting high-quality DNA facilitates efficient examination of fetal DNA sequences. © 2006 The American College of Medical Genetics.

publication date

  • October 1, 2006

Digital Object Identifier (DOI)

start page

  • 615

end page

  • 619

volume

  • 8

issue

  • 10