ABCMdb

PMID: 17890437

Montgomery J, Wittwer CT, Kent JO, Zhou L
Scanning the cystic fibrosis transmembrane conductance regulator gene using high-resolution DNA melting analysis.
Clin Chem. 2007 Nov;53(11):1891-8. Epub 2007 Sep 21., [PubMed]

Sentences

No. Mutations Sentence Comment

8 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Arg553* The melting patterns of most heterozygotes were unique (37 of 40 pairs within the same amplicon), the exceptions being p.F508del vs p.I507del, p.G551D vs p.R553X, and p.W1282X vs c.4002A>G. Login to comment 9 ABCC7 p.Gly542* The homozygotes p.G542X, c.2789 ؉ 5G>A, and c.3849 ؉ 10kbC>T were directly identified, but homozygous p.F508del was not. Login to comment 17 ABCC7 p.Trp1282* Several common mutations are specific to ethnic groups, such as p.W1282X among Ashkenazi Jews and c.3120 ϩ 1GϾA among native Africans. Login to comment 90 ABCC7 p.Cys1395Tyr ABCC7 p.Leu1388Val ABCC7 p.Ser158Thr Four variants (p.S158T, c.3891GϾA, p.L1388V, and p.C1395Y) have not been reported previously, and each was detected in only a single sample. Login to comment 98 ABCC7 p.Arg75Gln ABCC7 p.Arg668Cys ABCC7 p.Gly576Ala ABCC7 p.Leu967Ser ABCC7 p.Cys1395Tyr ABCC7 p.Leu1388Val ABCC7 p.Ser158Thr Allele fraction (%) 1 125GϾC 3.8 3 356GϾA R75Q 3.5 4 605GϾC S158T Ͻ0.4b,c 6b 1001 ϩ 11CϾT 13.1 10 1540AϾG M470V 30.0d 1716GϾA 1.5 12 1859GϾC G576A 1.5 13 2134CϾT R668C 1.5 14a 2694TϾG 26.2 14b 2752 - 6TϾC 0.4 15 3032TϾC L967S 0.8 17b 3417AϾT T109S 1.5 19 3601 - 17TϾC 0.4 20 3891GϾA Ͻ0.4b,c 4002AϾG 1.5 21 4029AϾG 0.4 23 4294CϾG L1388V 0.4b 4316GϾA C1395Y 0.4b 4374 ϩ 13AϾG 0.4 24 4404CϾT 0.8 4521GϾA 20.8 a All variants were identified by scanning random panels and confirmed by sequencing. Login to comment 107 ABCC7 p.Gly85Glu p.G85E is in a region of lower stability and the change in melting curve shape is most distinctive at lower temperatures. Login to comment 108 ABCC7 p.Arg75* In contrast, the p.R75X variant occupies a region of greater thermal stability and exhibits a shape change at higher temperatures. The disease-associated variants tested in the blinded study are shown in Table 2. Login to comment 127 ABCC7 p.Gly85Glu ABCC7 p.Arg75* G85E is in a low-temperature domain compared with R75X, resulting in deviation of the melting curves at different temperatures. The PCR product was 169 bp in length. Login to comment 132 ABCC7 p.Gly542* Furthermore, 3 homozygous disease-causing variants were identified in the blinded study: p.G542X, c.2789 ϩ 5GϾA, and c.3849 ϩ 10kbCϾT. Login to comment 141 ABCC7 p.Gly551Asp ABCC7 p.Arg553* Each genotype traces a different path, with the exception of p.G551D and p.R553X. Login to comment 145 ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Trp1282* ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Asn1303Lys ABCC7 p.Arg1162* ABCC7 p.Ile148Thr ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Arg31Cys ABCC7 p.Arg75* 2 223CϾT R31C 3 355CϾT R75X 386GϾA G85E 4 482GϾA R117H 575TϾC I148T 621 ؉ 1GϾTb 5 711 ؉ 1GϾT 7 1078delT 1132CϾT R334W 1150delA 1172GϾC R347P 8 1341 ϩ 18AϾCc 9 1496CϾA A455E 10 1651-1653del I507del 1653-1655del F508deld 11 1717 - 1GϾA 1756GϾT G542Xe 1784GϾA G551Db 1789CϾT R553Xf 1811GϾC R560T 12 1898 ؉ 1GϾA 13 2184delA 14b 2789 ؉ 5GϾAe 16 3120 ؉ 1GϾA 18 3500 - 2AϾTg 19 3616CϾT R1162X 3659delC Intron 19 3849 ؉ 10kbCϾTe 20 3978GϾA W1282X 21 4041CϾG N1303K 22 4178GϾA G1349Dc a Disease-causing variants recommended for genotyping by the ACMG (4) are in bold. Login to comment 148 ABCC7 p.Gly1349Asp c c.1341 ϩ 18AϾC was associated with the variant p.G1349D in 1 sample. Login to comment 162 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Arg553* The exceptions included 3-bp deletions, p.I507del/p.F508del, and 2 single-base variant pairs (p.G551D/p.R553X and p.W1282X/ c.4002AϾG). Login to comment 165 ABCC7 p.Gly551Asp ABCC7 p.Arg553* Fig. 4 in the online Data Supplement details an unlabeled probe assay to differentiate p.G551D from p.R553X. Login to comment 166 ABCC7 p.Trp1282* Fig. 5 in the online Data Supplement describes a small amplicon assay to distinguish the disease-causing variant p.W1282X from the benign variant c.4002AϾG. Login to comment 202 ABCC7 p.Gly551Asp ABCC7 p.Gly542* With the exception of p.G551D and p.R553X, the melting profile of each genotype is distinct, including the G542X homozygote. Login to comment 205 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* The substitutions were 5 bp (p.G551D and p.R553X) and 24 bp (p.W1282X and c.4002AϾG) apart, and the variants within each pair were of the same type (16). Login to comment