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Clin Chem. 2015 Dec;61(12):1515-23. doi: 10.1373/clinchem.2015.244962. Epub 2015 Oct 14.

Cell-Free RNA Is a Reliable Fetoplacental Marker in Noninvasive Fetal Sex Determination.

Author information

1
Department of Clinical Genetics and GROW School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, the Netherlands;
2
Department of Human Genetics, Radboud University Medical Center, Nijmegen, the Netherlands.
3
Department of Clinical Genetics and.
4
Department of Clinical Genetics and GROW School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, the Netherlands; Department of Human Genetics, Radboud University Medical Center, Nijmegen, the Netherlands. joris.veltman@mumc.nl.

Abstract

BACKGROUND:

Noninvasive genetic tests that use cell-free fetal DNA (cffDNA) are used increasingly in prenatal care. A low amount of cffDNA can have detrimental effects on the reliability of these tests. A marker to confirm the presence of fetal nucleic acids is therefore required that is universally applicable and easy to incorporate.

METHODS:

We developed a novel multiplex, single-tube, noninvasive fetal sex determination assay by combining amplification of AMELY cffDNA with one-step reverse transcription (RT)-PCR of trophoblast-derived cell-free RNA (cfRNA), which functions as a sex-independent fetoplacental marker. We tested plasma samples from 75 pregnant women in duplicate in a blinded fashion. The fetus was considered to be male in the case of a positive result for AMELY and cfRNA amplification in both RT-PCRs. The fetus was considered to be female in the case of negative AMELY and positive cfRNA result in both RT-PCRs. In other cases, the test was repeated. We compared the results with invasive prenatal testing and pregnancy outcomes.

RESULTS:

The AMELY cffDNA amplification and cfRNA result was unambiguous and identical in duplicate in 71 of 75 plasma samples (95%). Four samples (5%) required an extra replicate because of an absent fetoplacental marker. Thereafter, fetal sex was correctly determined in all 75 plasma samples.

CONCLUSIONS:

Amplification of trophoblast-derived cfRNA is a reliable marker for the confirmation of the presence of fetoplacentally derived nucleic acids in noninvasive fetal sex determination.

PMID:
26467504
DOI:
10.1373/clinchem.2015.244962
[Indexed for MEDLINE]
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