Cell-based non-invasive prenatal diagnosis in a pregnancy at risk of cystic fibrosis

TY - JOUR

T1 - Cell-based non-invasive prenatal diagnosis in a pregnancy at risk of cystic fibrosis

AU - Jeppesen, Line Dahl

AU - Hatt, Lotte

AU - Singh, Ripudaman

AU - Ravn, Katarina

AU - Kølvraa, Mathias

AU - Schelde, Palle

AU - Uldbjerg, Niels

AU - Vogel, Ida

AU - Lildballe, Dorte L

N1 - © 2020 John Wiley & Sons Ltd.

PY - 2021/1

Y1 - 2021/1

N2 - OBJECTIVE: We aimed to develop cell-based NIPT for cystic fibrosis (CF) and test a pregnancy at risk of two common pathogenic variants.METHOD: A pregnant woman carrying monozygotic twins opted for prenatal testing as she and her partner were heterozygote carriers of F508del (c.1521:1523del). The partner was also positive for the CFTR-related variant R117H (c.350G>A). Fetal trophoblasts from maternal blood were enriched and isolated using antibodies and a capillary-based cell-picking instrument. Multiplex PCR-based fragment length analysis was performed on the extracted fetal DNA for STR-genotyping, fetal gender and F508del variant status. The R117H variant status was tested using SNaPshot analysis.RESULTS: The fetal origin of the isolated cells was verified by detection of two paternally inherited STR alleles and an Y chromosome marker, while no maternal DNA contamination was detected. The direct variant analysis detected F508del heterozygosity and the SNaPshot analysis for R117H detected only the normal allele. Thus, the results showed that the fetuses were healthy carriers of F508del, concordant with the findings of conventional prenatal testing.CONCLUSION: Cell-based NIPT could accurately state the fetal variant status and distinguish fetal trophoblasts from maternal cells. In the future, cell-based NIPT may provide an accurate less invasive alternative to chorionic villous sampling.

AB - OBJECTIVE: We aimed to develop cell-based NIPT for cystic fibrosis (CF) and test a pregnancy at risk of two common pathogenic variants.METHOD: A pregnant woman carrying monozygotic twins opted for prenatal testing as she and her partner were heterozygote carriers of F508del (c.1521:1523del). The partner was also positive for the CFTR-related variant R117H (c.350G>A). Fetal trophoblasts from maternal blood were enriched and isolated using antibodies and a capillary-based cell-picking instrument. Multiplex PCR-based fragment length analysis was performed on the extracted fetal DNA for STR-genotyping, fetal gender and F508del variant status. The R117H variant status was tested using SNaPshot analysis.RESULTS: The fetal origin of the isolated cells was verified by detection of two paternally inherited STR alleles and an Y chromosome marker, while no maternal DNA contamination was detected. The direct variant analysis detected F508del heterozygosity and the SNaPshot analysis for R117H detected only the normal allele. Thus, the results showed that the fetuses were healthy carriers of F508del, concordant with the findings of conventional prenatal testing.CONCLUSION: Cell-based NIPT could accurately state the fetal variant status and distinguish fetal trophoblasts from maternal cells. In the future, cell-based NIPT may provide an accurate less invasive alternative to chorionic villous sampling.

KW - DNA

UR - http://www.scopus.com/inward/record.url?scp=85096682347&partnerID=8YFLogxK

U2 - 10.1002/pd.5861

DO - 10.1002/pd.5861

M3 - Journal article

C2 - 33150588

SN - 0197-3851

VL - 41

SP - 234

EP - 240

JO - Prenatal Diagnosis

JF - Prenatal Diagnosis

IS - 2

ER -