ABCMdb

PMID: 26493493

Destouni A, Poulou M, Kakourou G, Vrettou C, Tzetis M, Traeger-Synodinos J, Kitsiou-Tzeli S
Single-cell high resolution melting analysis: A novel, generic, pre-implantation genetic diagnosis (PGD) method applied to cystic fibrosis (HRMA CF-PGD).
J Cyst Fibros. 2015 Oct 19. pii: S1569-1993(15)00217-9. doi: 10.1016/j.jcf.2015.09.009., [PubMed]

Sentences

No. Mutations Sentence Comment

79 ABCC7 p.Asp110His ABCC7 p.Arg334Gln Validation case no. Genotype combination (HGVS CFTR reference sequences NM000492.3 and NG016465.1) Light scanner (LSC) exons (Montgomery et al. 2007) Number of cells tested Amplicons in multiplex RxNa PCR efficiency (%) Overall ADO (%) 1 p.Arg334Gln and c.489+3ANG 7.2 and 4.2 48 8 98.85% 2% 2 p.Phe508del and p.Leu732X 10 and 13.3 60 8 98% 1.66% 3 p.Phe508del and p.Asp110His 10 and 4.1 60 8 96.60% 3.33% 4 p.Phe508del and p.Gly542X 10 and 11 40 8 98% 3% Total lymphocytes tested 208 a RxN: reaction, ADO: allele drop-out 3 The mutation p.Phe508del could otherwise more rapidly be detected by fluorescent fragment analysis by incorporating the relevant primer set in the first PCR. Login to comment 101 ABCC7 p.Asp1152His ABCC7 p.Glu826Lys ABCC7 p.Gly1069Arg ABCC7 p.Gly1069Arg ABCC7 p.Gly1069Arg ABCC7 p.Arg334Gln ABCC7 p.Glu279Asp ABCC7 p.Asp1312Gly PGD case no. Genotype combination (HGVS CFTR reference sequences NM000492.3 and NG016465.1) LSC exons (legacy nomenclature) (Montgomery et al., 2007) No. of blastomeres received for diagnosis No. of blastomeres amplified No. of blastomeres genotyped No. of unaffected blastomeres Pregnancy Confirmation of PGD result by PNDa 1 p.Arg334Gln and c.489+3ANG 7.2 and 4.2 5 4 4 2 No N/A 2 p.Phe508del and p.Phe508del 10 11 10 10 6 Yes Yes 3 p.Phe508del and c.489+1GNT 10 and 4.2 8 7 7 4 Yes Yes 4 p.Phe508del and p.Leu732X 10 and 13.3 10 8 8 5 Yes Yes 5 p.Phe508del and p.Asp1152His 10 and 18 4 3 3 3 Yes Yes 6 p.Phe508del and c.2051_2052delAAinsG 10 and 13.2 5 4 4 3 Yes Yes 7 p.Phe508del and p.Gly1069Arg 10 and 17bA1 9 9 9 4 Yes No (BP) 8 p.Phe508del and p.Gly542X 10 and 11 3 3 3 2 Yes Yes 9 p.Phe508del and c.3140-26ANG 10 and 17bA1 8 8 7 3 No N/A 10 p.Gly542X and p.Gly542X 11 2 2 2 1 No transfer 11 p.Glu826Lys and p.Phe508del 13.4 and 10 6 6 4 3 Yes Miscarried 12 p.D1312G and c.489+1GNT 21 and 4.2 11 9 9 8 Yes Miscarried 13 p.Glu279Asp and p.Phe508del 6b and 10 7 7 5 3 No N/A 14 p.Phe508del and p.Gly1069Arg 10 and 17bA1 9 8 8 5 Yes No (BP) 15 p.Glu822X and p.Gly1069Arg 13.4 and 17bA1 5 5 5 5 Yesb Miscarried Total 103 93 88 57 N/A: non-applicable, BP: biochemical pregnancy. Login to comment 147 ABCC7 p.Gly1069Arg Only 2 of the 15 clinical cases performed had the same genotype combination (p.Phe508del and p.Gly1069Arg) and only 1/15 where both parents carried p.Phe508del. Login to comment