Non-invasive prenatal diagnosis (NIPD) of cystic fibrosis: an optimized protocol using MEMO fluorescent PCR to detect the p.Phe508del mutation

Guissart, Claire; Dubucs, Charlotte; Raynal, Caroline; Girardet, Anne; Mau-Them, Frédéric Tran; Debant, V.; Rouzier, Cécile; Boureau-Wirth, Amandine; Haquet, Emmanuelle; Puechberty, Jacques; Bieth, Éric; Deguine, D. Dupin; Kien, Philippe Khau Van; Bréchard, Marie-Pierre; Pritchard, Victoria L.; Kœnig, M.; Claustres, Mireille; Vincent, Marie-Claire

doi:10.1016/j.jcf.2016.12.011

Cited by 19 publications

(11 citation statements)

References 19 publications

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“…Our results showed that indirect testing based on fluorescent multiplex PCR can be safely used for NIPD, reaching sensitivity and specificity similar to those seen in previous studies [10,21,23,24]. However, this test is less informative than the SNP-haplotype-based approach by next-generation sequencing [25][26][27][28][29].…”

Section: Discussionsupporting

confidence: 86%

“…Appendix B). The p.Phe508del mutation was correctly detected in all families' samples [10] as well as SRY sequence. We found that the number of informative markers was always higher in amniotic fluid or chorial villosities than in corresponding maternal plasma ( Fig.…”

Section: Clinical Validationmentioning

confidence: 84%

“…Previous studies were performed on small sample groups and on targeted mutation of the CFTR gene [7,9,10,[16][17][18][19][20]. In contrast, here we analyzed more than 14 samples from high-risk CF pregnancies and over 16 samples from pregnancies with a priori CF risk based on ultrasound findings, the latter secondarily excluded following parental genotyping.…”

Section: Discussionmentioning

confidence: 99%

“…material) in K3 EDTA Vacutainer tubes or cell-stabilizing tubes (Streck blood collection tube [BCT]). Samples were processed up to 24 h (EDTA tubes) or up to 72 h (BCT) after sampling as previously described [9,10].…”

Section: Participants and Processing Of Blood Samplesmentioning

confidence: 99%

“…All samples were tested with both fluorescent multiplex PCR assays (Plex A and Plex B). The Plex A set was performed as previously described [10]. The PCR mix for the Plex B set contained 15 μL of 2X multiplex PCR buffer (Platinium; Life Technologies, France) and 0.1-0.6 μM each of primer pairs (Table 1).…”

Section: Assay Designmentioning

confidence: 99%

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A Broad Test Based on Fluorescent-Multiplex PCR for Noninvasive Prenatal Diagnosis of Cystic Fibrosis

et al. 2018

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Background: Analysis of cell-free fetal DNA in maternal plasma is very promising for early diagnosis of monogenic diseases. However, it has been limited by the need to set up patient- or disease-specific custom-made approaches. Here we propose a universal test based on fluorescent multiplex PCR and size fragment analysis for an indirect diagnosis of cystic fibrosis (CF). Methods: The test, based on haplotyping, includes nine intra- and extragenic short tandem repeats of the CFTR locus, the coamplification of p.Phe508del (the most frequent mutation in CF patients worldwide), and a specific SRY sequence. The assay is able to determine the inherited paternal allele. Results: Our simple approach was successfully applied to 30 couples and provided clear results from the maternal plasma. The mean rate of informative markers was sufficient to propose it for use in indirect diagnosis. Conclusions: This noninvasive prenatal diagnosis test, focused on indirect diagnosis of CF, offers many advantages over current methods: it is simple, rapid, and cost-effective. It allows for the testing of a large number of couples with high risk of CF, whatever the familial mutation of the CFTR gene. It provides an alternative method to reduce the number of invasive tests.

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