Authors
Nancy B. Y. Tsui, Rossa W. K. Chiu, Chunming Ding, and Y. M. Dennis Lo
Summary
Noninvasive prenatal diagnosis is an actively researched area in medical genetics. The discovery of cell-free fetal DNA in maternal plasma has opened up a new source of fetal genetic materials for molecular analysis1. The finding that circulating fetal RNA, in addition to fetal DNA, released by the placenta2, is present in maternal plasma has further increased the potential diagnostic applications of plasma nucleic acid analysis. We have recently demonstrated that allelic ratio analysis of single nucleotide polymorphisms (SNPs) present in such circulating placental mRNA would allow us to determine, with high sensitivity and specificity, the aneuploidy status of the fetus noninvasively3. Here, we describe in detail a protocol for carrying out such an analysis, including the blood processing steps, RNA extraction from plasma, RNA quantification by real-time quantitative reverse-transcriptase polymerase chain reaction (QRT-PCR), and allelic ratio determination by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In this protocol, the PLAC4 gene, which is located on chromosome 21, is used as an example3. The allelic ratio determination for PLAC4 in maternal plasma can therefore be used as a noninvasive method for detecting trisomy 21.Further details
The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.Advertisement
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